ABSTRACT

This study was carried out to investigate the effects of adding fruit of juniper berry (Juniperus communis) on performance, some carcass characteristics, serum parameters, histopathological features and jejunum villi lengths in broiler diets. A total of 150 male chicks were used in the study. The chicks were housed for 42 days. One of the experimental groups was a control group, and the diets of the 4 treatment groups included 0.5%, 1%, 1.5% and 2% dried natural juniper berries, respectively. Addition of fruit of juniper berry to the diets had positive effects on body weight, average daily weight gain, feed conversion rate and slaughter weight, feed intake, hot carcass yield and serum calcium and phosphorus levels. Juniper berry addition in the diets also increased the length of the jejunum villi. As a result, it can be recommended to add up to 1% of juniper berry in broiler diet.

Keywords:
Broiler; Juniper berry; Performance; Villi lengths

INTRODUCTION

Juniperus ssp. genus is known to have about 60 species worldwide (Kesbic, 2019Kesbic OZ. Effects of juniper berry oil on growth performance and blood parameters in common carp (Cyprinus carpio). Aquaculture 2019;50:342-349.). Juniper berry contains up to 2% volatile oil comprising of the main compounds α-pinenes (35.4-51.85%), limonene (1.2-25.1%) and β-caryophyllene [1.3-4.2%) with smaller amounts of cadinene, α-terpinenes, tripionela, O-cymene, isopinocarveal, cedrene, elemene, thiopsene (Pandey et al., 2018Pandey S, Tiwari S, Kumar A, Niranjan A, Chand J, Lehri A, et al. Antioxidant and anti-aging potential of juniper berry (Juniperus communis L.) essential oil in Caenorhabditis elegans model system. Industrial Crops and Products 2018;120(15):113-122.). Juniper berry has been reported to have appetizing, stress preventing, immune system boosting, anti-indigestive, analgesic, hepatoprotective, antihyperlipidemic, antimicrobial, anti-inflammatory, diuretic, antioxidant, reactive oxygen species preventing, anti hypercholesterolemic, anticalaleptic activating and neuroprotective activities. It was determined that the extract obtained from Juniperus communis of fruits decreased the glycemic level of blood (Loziene and Venskutonis, 2016Loziene K, Venskutonis PR. In essential oils in food preservation, flavor and safety. Preedy V, editor. Juniper (Juniperus communis L.) oils. London: Academic Press 2016; p.495-500.; Pandey et al., 2018; Kesbiç, 2019). Although studies are limited in poultry, there are many studies on the use of essential oils derived from medicinal and aromatic plants (Cengiz et al., 2015Cengiz SS, Yesilbag D, Meral Y, Çetin I, Biricik H. Juniper oil improves oxidative stability and eggshell and albumin quality of quail eggs. British Poultry Science 2015;56(1):58-65.; Okoro, 2016Okoro MC. Effect of dietary inclusion of ginger (Zingiber officinale) on performance of broilers and laying chickens [thesis]. Owerri (NG): Federal University of Technology; 2016.; Sevim and Cufadar, 2017Sevim B, Cufadar Y. Effects of an addition of different essential oils and their combinations to diets on performance and carcass characteristics parameters in broilers. Turkish Journal of Agriculture-Food Science and Technology 2017;5(8):964-968.). Inci et al. (2016Inci H, Gökçe Ö, Sengül AY, Sögüt B, Nursoy H, Sengül T. Using juniper berry (Juniperus communis) as a supplement in Japanese quail diets. Revista Brasileira de Zootecnia 2016;45(5):230-235.) reported that the addition of juniper berry at 0.5-1% level had positive effects on fattening performance and some carcass characteristics in the quail diet. However, no studies have been found showing the use of juniper berry directly in broilers rations. Therefore, Iin this study, we investigated the effects of adding 0.5%, 1%, 1.5% and 2% concentration level of Juniper berry to the broiler diets on body weight, daily gain, feed intake, feed efficiency, and parameters of serum, carcass and histopathology and jejunum villi lengths. This study was carried out to draw attention to the use of juniper berries, which are abundant in nature, in broiler rations and to shed light on future studies.

MATERIALS AND METHODS

Animal and feed material and experimental design

150 three-day-old chicks of Ross-308^® used in the experiment were obtained from CP Pullet Hatchery^® in Erzincan, Turkey. The experiment was started after the approval of protocols by the Local Ethical Committee of Van Yüzüncü Yıl University affiliated in Van, Turkey (Date: 02.11.2009, Decision No: 07). The chicks were distributed into 5 groups with 30 animals in each group. The experiment was carried out at Van Yüzüncü Yıl University Animal Research and Application Farm Directorate in Turkey (42º40’E and 37o43’). The chicks were housed in 1x1.5 m compartments with 10 chicks and under 23 hours of light followed by 1 hour darkness of photoperiods per daily cycle. One type of diet was used during the experiment, it was arranged according to the NRC (1994NRC. The nutrient requirements of poultry. 9th ed. Washington: National Academy Press; 1994.) (Table 1). In the control group (C) and 4 groups which were formed by 0.5%, 1%, 1.5% and 2% dried natural berries of Juniper communis juniper berries added to the diets. Feeds and water were fed ad libitum to chicken.

Feed, performance, serum, carcass and histopathologic of analysis

Feed samples were obtained at 105 °C and 16 h in oven-drying, ash for 2 h at 600 °C with furnace, fat by diethyl ether extraction method crude protein by N×6.25 Kjeldahl method (AOAC, 1990). Body weight, daily body weight gain or average daily gain (ADG), feed intake and feed conversion rate (FCR) of all birds used in the experiment were determined weekly. At the end of the experiment, 7 chickens from each group were slaughtered, and serums were removed from their blood. The values of alanine amino transaminase (ALT), aspartate amino transaminase (AST), calcium (Ca), phosphorus (P), potassium (K), chlorine (Cl) and sodium (Na) were analyzed on the auto analyzer (Cobas Integra-800 Analyzer, Roche Diagnostics GmbH, Mannheim, DE) in Van Medical Park Hospital. The birds’ heads, feet, feathers and organs of digestive system except liver and kidney were separated from those trunks as “hot carcass”. Division of hot carcass weight by body weight before slaughtering was calculated as “carcass yield”. Fats of covers of gizzard, duodenum and intestine were weighed as “abdominal fat” and abdominal fat: hot carcass rate as “abdominal fat rate” (Nursoy et al., 2011Nursoy H, Sögüt B, Tasal T, Aldemir R, Kaplan O, Altaçli S. The effects of varying dietary Na/K ratio and electrolyte balance of diets on growth, blood gases, hematological variables, ionized calcium and carcass traits in broiler chickens. Journal of Veterinary Faculty of Kafkas University 2011;17(6):979-986.). Liver and jejunal tissues were taken into a buffered formalin solution 10% and then routed to paraffin blocks procedures. Sections of 4µm thickness from each block were stained with hematoxylin-eosin and immunohistochemistry and examined by light microscopy and pictures were taken (Apaydın Yıldırım et al., 2017; Ertekin et al., 2017).

Statistical analysis

Data obtained were analyzed by SAS statistical program according to one-way ANOVA. The following mathematical model was applied:

in which Yij represents the j-th observation on the i-th level of juniper berry content (i=0, 0.5, 1, 1.5, and 2% of the diet), µ is the overall mean response, τi is the effect due to the i-th level of juniper berry content, and εij represents the random error present in the j-th observation on the i-th treatment. The differences between the groups were performed by Duncan multiple comparison test within SAS 9.2 (SAS, 2008).

RESULTS

In general, live weights of birds were found to be high between 0-6 weeks in the control group and in 0.5-1% groups and the highest body weight was determined in 3-6 weeks and 1-6 weeks in the 0.5% group (p<0.05). Likewise, 0.5% group had the highest ADG. The lowest daily feed intake was determined in the 0.5% group between 3-6 weeks (p<0.01) and in the 0.5% group during the trial (Table 2).

The slaughter weight on day 42 was found to be highest in 0.5% group (p<0.05). There was no statistically significant difference between the groups with hot carcass weight, abdominal fat weight and abdominal fat rate (p>0.05), (Table 3). Serum parameters of control and treatment groups did not statistically affect the serum ALT, AST, Cl, K and Na values of juniper fruits included in the diet (p>0.05, Table 4). However, serum P and Ca levels were the highest in 0.5% group (p<0.01). Histopathological features were not affected by the addition of juniper fruit to the chicken diet on hepatocytes, sinusoidal cavities, mucosal, submucosa, and serosa of the liver and jejunal tissues (Figure 1 and Figure 2). The juniper groups 1% and 2% had a villi length higher than the control group (p<0.01, Table 5). In addition, the lengths of juniper groups showed a higher tendency as the consecutive fruit increase in diets increased linearly (Graph 1).

DISCUSSION

In the present study, the effects of whole, dry and grounded portions of varying concentration levels of juniper fruits were investigated on different parameters during 6 weeks. The highest body weight was found at the control group in 1-3 weeks and the highest body weight was determined at the 0.5% group in 3-6 weeks and 1-6 weeks (p<0.05, Table 2). The highest ADG was found in the control group in 1-3 weeks, and 0.5% juniper fruit group was found to have the highest ADG in 3-6 weeks and 1-6 weeks (p<0.05). The positive effects of juniper berry were not observed on chickens in 1-3 weeks. However, using 0.5-1% of juniper berry in the broiler diet increases body weight and ADG of 3-6 weeks. Body weights and ADG of the groups in this study are in line with the results of Lewis et al. (2003Lewis MN, Rose SP, Mackenzie M, Tucker LA. Effect of dietary inclusion of plant extracts on the growth performance of male broiler chickens. British Poultry Science 2003;44(Supl 1):43-44.), Lee et al. (2003Lee KW, Everts H, Kappert HJ, Frehner M, Losa R, Beynen AC. Effects of dietary essential oil components on growth performance, digestive enzymes and lipid metabolism in female broiler chickens. British Poultry Science 2003;44(3):450-457.) and Yeşilbağ et al. (2014). The highest feed intake was found at 2% juniper group in 1-3 weeks (p<0.05) and 1-6 weeks (p<0.01) in the experiment. Feed intakes of data from the present study confirmed those values of Inci et al. (2016Inci H, Gökçe Ö, Sengül AY, Sögüt B, Nursoy H, Sengül T. Using juniper berry (Juniperus communis) as a supplement in Japanese quail diets. Revista Brasileira de Zootecnia 2016;45(5):230-235.). In contrast, they were not compatible with studies which showed that different aromatic plant or essential oils had no effects on BW and feed intake in broilers (Zeng et al., 2015Zeng Z, Zhang S, Wang H, Piao X. Essential oil and aromatic plants as feed additives in non-ruminant nutrition: a review. Journal of Animal Science Biotechnology 2015;6(1):2-10.; Bozkurt et al., 2016Bozkurt M, Ege G, Aysul N, Aksit H, Tüzün AE, Küçükyilmaz K, et al. Effect of anticoccidial monensin with oregano essential oil on broilers experimentally challenged with mixed Eimeria spp. Poultry Science 2016; 95:1858-1868.; Simitzis, 2017Simitzis PE. Enrichment of animal diets with essential oils-a great perspective on improving animal performance and quality characteristics of the derived products. Medicines 2017;4(2):2-21.).

Figure 1
A: Control group; liver tissues in normal histological structure, B: 1% group; liver tissues in normal histological structure, C: 2% group; liver tissues in normal histological structure, H&E, Bar:20µm.

Figure 2
A: Control group; jejunum tissues in normal histological structure, B: 1% group; jejunum tissues in normal histological structure, C: 2% group; jejunum tissues in normal histological structure, H&E, Bar: 100 µm.

When values of the FCR were examined in Table 2, the best utilization was determined at 0.5% group between 3-6 weeks (p<0.01) and 1-6 weeks (p<0.05). The results were supported by the values of Inci et al. (2016Inci H, Gökçe Ö, Sengül AY, Sögüt B, Nursoy H, Sengül T. Using juniper berry (Juniperus communis) as a supplement in Japanese quail diets. Revista Brasileira de Zootecnia 2016;45(5):230-235.) that a low level of juniper added to diet increased FCR. On the other hand, other researchers (Zeng et al., 2015Zeng Z, Zhang S, Wang H, Piao X. Essential oil and aromatic plants as feed additives in non-ruminant nutrition: a review. Journal of Animal Science Biotechnology 2015;6(1):2-10.; Bozkurt et al., 2016Bozkurt M, Ege G, Aysul N, Aksit H, Tüzün AE, Küçükyilmaz K, et al. Effect of anticoccidial monensin with oregano essential oil on broilers experimentally challenged with mixed Eimeria spp. Poultry Science 2016; 95:1858-1868.; Simitzis, 2017Simitzis PE. Enrichment of animal diets with essential oils-a great perspective on improving animal performance and quality characteristics of the derived products. Medicines 2017;4(2):2-21.) reported that different aromatic plant or essential oils in broiler diets had no positive effects on FCR.

The 0.5% group was found to have the highest slaughter weight after 42 days (p<0.05, Table 3). There was no statistical difference between the groups with hot carcass weight (p>0.05). Carcass yield was the lowest at 0.5% group (p<0.01). However, few researchers (Denli et al., 2004Denli M, Okan F, Uluocak AN. Effect of supplementation of herb essential oils on the growth performance, carcass and intestinal characteristics of quail (Cotunix coturnix japonica). South African Journal of Animal Science 2004;34(3):174-179.; Ocak et al., 2008Ocak N, Erener G, Ak FB, Sungu M, Altop A, Özmen A. Performance of broilers fed diets supplemented with dry peppermint (Mentha piperita L.) or thyme (Thymus vulgaris L.) leaves as growth promoter source. Czech Journal of Animal Science 2008;53(4):169-175.; Vazquez et al., 2017Vázquez RS, Macías JAG, Meléndez LAD, Hume ME, Zamora GM. Mexican orégano (Lippia berlandieri Schauer) oil on turkey slaughter quality. Ecosistemas Recursos Agropecuarios 2017;4(10):177-182.) reported that different aromatic plant or essential oils did not affect hot and cold carcass characteristics. The lowest values of both abdominal fat weight (26.57 g) and abdominal fat rate (1.46%) were determined in the 2% group, but no statistical significance was found (p>0.05) in the experiment. Similar results of decreased abdominal fat weight were also reported by Ocak et al. (2008) and Denli et al. (2004).

The serum parameters of the control and treatment groups did not statistically affect the serum ALT, AST, Cl, K and Na values of the juniper fruit included in the diet (p> 0.05, Table 4). However, serum P and Ca levels were found the highest in the 0.5% group (p<0.01). Histopathological features were not affected by adding juniper berry in the broiler diet on hepatocytes, sinusoidal cavities, mucosal, submucosa and serosa of the liver and jejunum tissues (Figure 1 and Figure 2). The villi lengths of the juniper groups of 1% and 2% were higher than those of the control group (p<0.01, Table 5). In addition, the villi lengths of the juniper groups showed a higher trend with increasing juniper berry concentrations in the diets as linear effects (Graph 1). The increase in digestion and absorption of nutrients stem from increased villi lengths leading to positive effect on body weight, ADG, feed intake and FCR. Since there were very limited studies on the use of juniper berry in broilers or poultry diets, the performance, serum parameters and histopathological features data could not be compared one-to-one.

Graph 1
Jejunum villus lengths (µm) of the groups.

CONCLUSION

It was determined in this study that adding juniper berry in broiler diet had positive effects on body weight, ADG, feed intake, FCR, carcass characteristics, serum parameters and histopathological features. For optimum benefits, up to 1% of juniper berry can be added in broiler diet.

REFERENCES

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