The Effect of Dietary Garlic (Allium Sativum), Black Cumin (Nigella Sativa) and Their Combination on Performance, Intestine Morphometry, Serum Biochemistry and Antioxidant Status of Broiler Chickens

I Aydogan E Yildirim A Kurum D Bolat M Cinar M Basalan A Yigit About the authors

ABSTRACT

This study was conducted to examine the effects of garlic (G), black cumin (BC) and G+BC within the broiler rations, on performance, relative organ weights, intestinal morphology, serum biochemistry and plasma total oxidant/ antioxidant status. Two hundred broilers chicks (Ross 308) were used and they were divided into 4 groups consisting of 5 subgroups of 10 animals each. The trial was ended on the 35th day. No addition was made to the control group ration. 5g / kg garlic (G), 5g / kg black cumin (BC) and 5g / kg G + 5g / kg BC were added to the experimental group rations, respectively.

There was no statistical difference among the trial groups in terms of BWG, FI, FCR and relative organ weights (p>0.05). Serum total protein (p<0.05) and albumin (p<0.001) levels were higher in BC and BC +G groups as compared to G and control group. Serum AST activity were lower in G group as compared to control and other experimental groups (p<0.05). On histological examinations, regarding the duodenal epithelium; there was no difference among the trial groups. Hyperemia was observed, especially in the G group, in macroscopic exams of lamina propria and other viticular areas. The lymph follicles were more wide spread in the G + BC, G and BC groups than in the control. The results of the study showed that the combination of BC and G can have beneficial effects, and different doses of G and BC may be used to see positive or negative effects.

Keywords:
Antioxidant status; Black Cumin; Broiler performance; Garlic; Serum biochemistry

INTRODUCTION

Poultry is one of the most important sources of food industry. Broiler chickens can be made ready to the market in six weeks time. Use of growth promoter was common in poultry industry to improve the performance (Apata, 2009Apata DF. Antibiotic resistance in poultry. International Journal of Poultry Science 2009; 8(4), 404-408.). Medicinal plants have been widely used for treatment of diseases in humans for centuries. Plant species and products have been used as feed additives in animal nutrition in recent years as well. Feed additives derived from plants are more preferable in animal production since they are natural, residue free, and less toxic compared to other synthetic feed additives (Wang et al., 1998Wang R, Li D, Bourne S. Can 2000 years of herbal medicine history help us solve problems in the year 2000. In Alltechs annual symposium 1998; Vol. 14, pp. 168-184.; Guo, 2003Guo F. Mushroom and herb polysachariides as alternative for antimicrobial growth promotors in poultry. 2003.). Herbal products such as garlic (Allium sativum) and black cumin (Nigella sativa), which are phytogenic feed additives, have been considered as an alternative to growth factors in poultry nutrition in the recent years.

Garlic (Allium sativum) has a variety of organasulphur containing compounds, such as allicin that has antibacterial, antifungal, antiparasite, antiviral, antioxidant, antihyperlipedemic and immunostimulatory effects (Yoo et al., 2014Yoo M, Lee S, Kim S, Hwang JB, Choe J, Shin D. Composition of organosulfur compounds from cool-and warm-type garlic (Allium sativum L.) in Korea. Food Science and Biotechnology 2014; 23(2):337-344.; Kodera et al., 2017Kodera Y, Ushijima M, Amano H, Suzuki JI, Matsutomo T. Chemical and Biological Properties of S-1-Propenyl-L-Cysteine in Aged Garlic Extract. Molecules, 2017; 22(4):570., Yıldırım & Çınar, 2017). Beneficial effects of garlic was shown on the performance of the broilers (Rehman & Munir, 2015Rehman Z, Munir MT. Effect of garlic on the health and performance of broilers. Veterinaria 2015; 3(1): 32-39.). However Konjufca et al. (1997Konjufca VH, Pesti GM, Bakalli RI. Modulation of cholesterol levels in broiler meat by dietary garlic and copper. Poultry science 1997; 76(9):1264-1271.) suggested that the addition of garlic in levels of 1.5, 3 and 4.5% was not effective on broiler performance. Kansal et al. (2017Kansal A, Ali N, Fahim A, Bharti M, Chandra G, Siddiqui R. Effect of Dietary Supplementation of Garlic on the Haemato-biochemical Parameters and Performance of Broiler Chickens. International Journal of Livestock Research 2017; 7(10): 223-230.) declared that 0.75% garlic in the mixed diet has a positive effect on biochemical parameters in broilers.

Black cumin (Nigella sativa) is a medicinal plant that belongs to the Ranunculacea family. It is a biologically effective substance with a healing potential and showed to be effective in the treatment of diabetes and cancer, has a diuretic effect and prevents high blood pressure. It has analgesic, antimicrobial, anthelmintic, bronchodilator, stomach protector, liver protector, anti-inflammatory effects (Yıldırım & Çınar, 2017). Black cumin seeds contain significant levels of protein, energy and essential oils. Black cumin’s nutritional composition contains protein and amino acids (22.7%), fat (38.20%), and total carbohydrates (31.94%). The major components of BC are the volatile oils thymoquinoline and dithymoquinoline (Zahoor et al., 2004Zahoor A, Ghafoor A, Aslam M. Nigella sativa-a potential commodity in crop diversification traditionally used in healthcare. Project on introduction of medicinal herbs and spices as crops. Ministry of Food. Agriculture and Livestock Education 2004; 280-295.). N. sativa seeds in feed serve as a growth promoter and improve broiler performance. Broilers fed with 1.5% crushed black seed obtained better performance and FCR (Al-Beitawi et al., 2009Al-Beitawi NA, El-Ghousein SS, Nofal AH. Replacing bacitracin methylene disalicylate by crushed Nigella sativa seeds in broiler rations and its effects on growth, blood constituents and immunity. Livestock Science 2009;125(2-3): 304-307.). Al Homidan et al. (2002Al-Homidan A, Al-Qarawi AA, Al-Waily SA, Adam SEI. Response of broiler chicks to dietary Rhazya stricta and Nigella sativa. British Poultry Science 2002; 43(2): 291-296.) found no negative effect of using 20 and 100 g/kg N. sativa seed on the growth broiler chickens. Saeid et al. (2013Saeid JM, Mohamed AB, AL-Baddy MA. Effect of adding garlic powder (Allium sativum) and black seed (Nigella sativa) in feed on broiler growth performance and intestinal wall structure. Journal of Natural Sciences Research 2013; 3(1): 35-41.) examined the combined effect of garlic powder and black cumin combination and found a beneficial effect on performance. Singh & Kumar (2018) studied black cumin in broilers and found that 1% and 1.5 % black cumin cause an increase in serum total protein and decrease in total cholesterol whereas the serum levels of glucose, calcium, phosphorus, uric acid, creatinine and serum ALT, GOT and ALP activities were unchanged. To our knowledge, as mentioned above, there are a lot of different results regarding the different doses of G or BC treatment in broilers. Therefore, the objective of this study was to determine the effect of 5 g/kg G, 5 g/kg BC and their combination on performance, some biochemical parameters, total antioxidant status and to examine resultant intestinal morphometry.

MATERIALS AND METHODS

Plant Material

Garlic and black cumin were obtained from a local market in Turkey.

Experimental design

A total of 200 one-day-old commercial male broiler chickens (Ross 308), were used in a 35-day experiment. The birds were weighed and randomly allocated to 4 treatment groups. Each group was divided into five replicates consisting of 10 chicks. Chicks were raised in stainless steel pens (40 × 65 × 98 cm). The ingredients and nutrient composition of diets given to chickens are presented in Table 1. Dietary treatments consisted of control group (basal diet) (C); the garlic group (basal diet + 5 g/kg G); black cumin group (basal diet + 5 g/kg BC) ; black cumin and garlic group (5 g/kg G + 5g/kg BC). The diets were analysed according to the AOAC (2000). Chicks were fed on ad libitum and clean drinking water throughout the study. The initial room temperature began at 33°C and was gradually reduced by 3°C per week until reaching 24°C.

Table 1
Ingredients and nutrient composition of the basal diet (%).

Performance Parameters and Weight of Organs

The birds were indivually weighed at weekly intervals (weeks 1 to 5). Average body weight (BW), body weight gain (BWG), feed intake (FI) and feed conversion ratio (FCR) were also calculated weekly (FI/ BWG). By the end of the trial, 3 chicks were randomly choosen from each pen; and in total 15 chicks were slaughtered, defeathered, processed (separation of head and feet), and removed (separation of digestive system). The liver, bursa of fabricius, spleen, and pancreas were excised, weighed and the relative organ weights were expressed as a percentage of BW. Animal care protocol was approved by the Kırıkkale University animal welfare Committee (Protocol Number: 2016/17).

Histological examination and Stereological analysis

In the study, 6 tissue samples with a length of 5 cm were taken from the duodenum of each animal. By applying the systematic random sampling rule (Gundersen and Jensen, 1987Gundersen HJG, Jensen EB. The efficiency of systematic sampling in stereology and its prediction. Journal of microscopy 1987; 147(3): 229-263.) to these pieces, 6 tissue samples with a length of 0.82 cm were obtained. Samples were kept in 10% neutral buffered formalin solution for 24 hours. Routine histological procedure was applied to all sections and then paraffin blocks were prepared. 40 µm thick and 6 µm thin sections were obtained from each paraffin block by microtome. All sections were stained with Crossman’s modified triple staining (Denk et al., 1989Denk W, Webb WW, Hudspeth AJ. Mechanical properties of sensory hair bundles are reflected in their Brownian motion measured with a laser differential interferometer. Proceedings of the National Academy of Sciences 1989; 86(14): 5371-5375.) for stereological and histological examination. The surface area of the duodenum in tissue samples was estimated by the Isotropic Fakir Method (Kubínová & Janácek, 1998Kubínová L, Janacek J. Estimating surface area by the isotropic Fakir method from thick slices cut in an arbitrary direction. Journal of Microscopy 1998; 191(2):201-211.). For Isotropic Fakir Method, a stereology system consisting of a microscope, Leica® DM4000, (Leica Mycrosystems CMS GMBH, Wetzlar, Germany), a computer controlled three axis stage (Ludl Mac 5000®; Ludl Electronic Products Ltd, NewYork), a digital camera (MBF® 2000R Fast 1394 Color; Qimaging, Surrey, Canada) and stereology software (Stereoinvestigator®; MBF Bioscience, Williston, VT) were used.

Blood Collection

At the end of the trial, blood samples were collected from the jugular vein of birds stunned before slaughter using heparinized and serum tubes and were centrifuged at 1600×g at 4 °C for 10 min and kept at -80 °C for serum biochemical analysis and plasma total oxidant (TOS) and total antioxidant (TAS) analysis.

Serum biochemistry

Serum AST, ALT activities, glucose, total cholesterol, high-density lipoprotein (HDL cholesterol), total bilirubin, total protein, albumin, üric acid, creatinine concentrations were assigned via an autoanalyzer, using diagnostic kits (Gesan Chem 400, Italy). Serum globulin levels were obtained by subtracting albumin values ​​from total protein values.

Analysis of Total Antioxidant and Oxidant Status

Total antioxidant status (TAS) was measured colorimetrically in plasma via an autoanalyser (Mindray BS300, China) commercially avaible kit (Rel Assay Diagnostic, Gaziantep, Turkey) according to the colorimetric method developed by Erel (2004Erel O. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clinical Biochemistry 2004; 37(4), 277-285.). This method was based on the oxidation of 2,2’-azino-bis (3ethylbenzotiazoline-6-sulfonic acid) radical to a colorless reduced form with the antioxidants present in the sample. As the calibration of the method was made by Trolox, the results were expressed as millimoles of trolox equivalentperliter (mmol trolox equiv/l).

Total oxidant status (TOS) was also analysed colorimetrically in plasma via an autoanalyser (Mindray BS300, China) a commercially avaible kit (Rel Assay Diagnostic, Gaziantep, Turkey). The method of the commercial kit was developed by Erel (2005Erel O. A new automated colorimetric method for measuring total oxidant status. Clinical biochemistry 2005; 38(12):1103-1111.). The procedure was based on the reduction of ferrous ion to ferric ion with the presence of various oxidative species in acidic conditions. The calibration was done by hydrogen peroxide, therefore the data expressed as micromolar hydrogen peroxide equivalent per liter (µmol H2O2 equiv/l).

Statistical Analysis

The data obtained from biochemical and performance analysis were given as mean ± standart error and were analyzed using SPSS 16.0 for windows (SPSS Inc., Chicago, IL, USA). A one-way analysis of variance (ANOVA) was done to determine the significance of differences among groups. Differences between means were analyzed using Duncan’s multiple range test. Statements of statistical significance were based on a probability ofp< 0.05.

RESULTS

BWG, FI and FCR of the groups were shown in Table 2. No statistically significant difference was found in the mean BWG, FI and FCR of the control and trial groups (G, BC and G + BC) (p>0.05). However at the end of the experiment, the groups that had garlic added to the diets showed numerical improvement in FCR.

Table 2
The effect of garlic and black cumin added to the rations on growth performance in broilers.

The findings regarding the various organ weights assessed at the end of the trial and their ratio to the percentage of BW (% BW) were presented in Table 3. There was no significant difference among trial groups (p>0.05).

Table 3
Effects of supplementation with Garlic and Black cumin on relative organ weights (% of body weight) and duodenum surface area (mm2) of the trail groups.

As indicated in Table 3, the duodenum surface area of control is 158.3 ± 14.44 mm2 (Mean ± SD), garlic group is 161.6 ± 5.35 mm2 (Mean ± SD), BC is 159.8 ± 11.25 mm2 (Mean ± SD) and G+BC is 144.8 ± 12.95 mm2 (Mean ± SD). There was no statistical difference among the trial groups (p>0.05). On histological examinations, in the duodenal epithelium; there was no difference in the G, BC and G+BC groups compared to the control (Figure 1 A-B). However, goblet cells were more concentrated in the crypts in the application groups (Figure 1 C). Hyperemia was noted, especially in the garlic group, in overlapping macroscopic findings in the lamina propria and other connective tissue areas (Figure 1 D). The lymph follicles were more wide spread in the G, BC and G+BC groups than in control (Figure 1 E-F)

The results of the serum biochemistry and plasma TAS and TOS were shown in Table 4. Aspartate amino transferase activity was significantly lower in the garlic group compared to other groups. Serum total protein (TP) was significantly increased in BC + G as compared to G and control groups, on the other hand the level of TP was higher in BC group comprared to garlic group (p<0.05). The groups that are given BC + G to the ration had significantly increased serum albumin (p<0.001) levels as compared to G, BC and control groups. The albumin level in BC group is significantly higher than the control and G group (p<0.001). Serum ALT activity, total cholesterol, HDL-cholesterol, total bilirubin, globulin, glucose, uric acid, creatinine concentrations were not affected by the dietary treatments (p>0.05). Also plasma TAS and TOS are not affected by the dietary treatments.

Table 4
Effects of Garlic and Black cumin supplementation on serum biochemistry and plasma TAS and TOS of broilers (n=13).

DISCUSSION

No statistically significant difference was found in the mean BWG, FI and FCR of the control and trial groups (G, BC and G+BC) at the end of the experiment (p>0.05). However, the garlic group showed numerical improvement in FCR. These results were consistent with Issa and Omar (2012Issa KJ, Omar JMA. Effects of garlic powder on performance and lipid profile of broilers. Open Journal of Animal Science 2012; 2: 62-68.), who reported that chickens fed with diet that had additional 0.2% and 0.4% garlic powder did not have a statistically significant change on body weight and body weight gain. Likewise, Lee et al. (2016Lee KW, Lee KC, Kim GH, Kim JH, Yeon JS, Cho SB, Chang BJ, Kim SK. Effects of dietary fermented garlic on the growth performance, relative organ weights, intestinal morfology, cecal microflora and serum characteristics of broiler chickens. Brazilian Journal of Poultry Science, 2016; 18 (3): 511-518.) showed that daily BWG, FI and FCR were not affected in the groups treated with fermented garlic (0.1%; 0.3%; 0.5%). However, Mahmood et al. (2009Mahmood S, Mushtaq-Ul-Hassan M, Alam M, Ahmad F. Comparative efficacy of Nigella sativa and Allium sativum as growth promoters in broilers. International Journal of Agriculture and Biology 2009; 11(6): 775-778.) reported that the addition of 0.5% garlic to the broiler ration improved body weight gain.

Although the rates of FI and FCR were numerically lower in the groups received garlic, the results were not found statistically significant (p>0.05). Adebiyi et al. (2017Adebiyi FG, Ologhobo AD, Adejumo IO. Efficacy of Allium sativum as growth promoter, immune booster and cholesterollowering agent on broiler chickens. Asian Journal of Animal Science 2017; 11(5): 202-213.) reported that there was no significant difference in FI and FCR of chickens fed on 1%, 2% and 3% raw and dry garlic. These results are compatible with our study. Contrary to the results of this study, Elagib et al. (2013Elagib HAA, El-Amin WIA, Elamin KM, Malik HEE. Effect of dietary garlic (Allium sativum) supplementation as feed additive on broiler performance and blood profile. Journal of Animal science advances 2013; 3(2): 58-64.) who used 0, 3 and 5% garlic powder and Borgohain et al. (2017Borgohain B, Mahanta JD, Islam R, Sapcota D, Sarma S, Borah MC. Effect of feeding garlic (Allium sativum) as prebiotic on the performance of broiler chicken. International Journal of Livestock Research 2017; 7(7): 225-233.) that used garlic powder at the rate of 0.5, 1.0 and 1.5% reported an increase in feed intake and an improvement in feed utilization in the diets containing garlic. These differences in results may be due to various factors such as the level of garlic used, the ration composition, and the agricultural situation. Ashayerizadeh et al. (2009Ashayerizadeh O, Dastar B, Shargh MS. Use of garlic (Allium sativum), black cumin seeds (Nigella sativa L.) and wild mint (Mentha longifolia) in broiler chicken diets. Journal of Animal and Veterinary Advances 2009; 8(9): 1860-1867.) observed that the addition of garlic powder, black cumin powder and wild peppermint to the broiler rations did not have a significant effect on feed intake, whereas the group that had garlic added to the diet had significantly improved body weight and FCR compared to the control.

Lewis et al. (2003Lewis MN, Rose SP, Mackenzie AM, Tucker LA. Effects of dietary inclusion of plant extracts on the growth performance of male broiler chickens. Spring Meetingof the WPSA UK Branch Posters 2003; 43-44.) examined the effects of high levels of garlic extract on the performance of broiler chickens for 7-27 days and stated that it increased the body weight by 7%. Saeid et al. (2013Saeid JM, Mohamed AB, AL-Baddy MA. Effect of adding garlic powder (Allium sativum) and black seed (Nigella sativa) in feed on broiler growth performance and intestinal wall structure. Journal of Natural Sciences Research 2013; 3(1): 35-41.) observed a significant improvement in the ratio of BW, BWG and FCR by the addition of 0.5% garlic powder, black cumin and combination to the ration. Mansoub and Mohammad (2011Mansoub NH, Mohannad AMN. Effect of garlic, thyme and yogurt compared to antibiotic on performance, immunity and some blood parameters of broiler chicken. Indian Journal of Animal Science 2011; 81(12): 1197-1200.) found an improvement in BW and FCR in the group in which 1 g / kg of garlic was added, compared to other groups.

Black cumin has a positive effect as well as a neutral effect on performance. Lymia et al. (2010Lymia HA, MajeedKhadiga A, Abdelati Nabiela M Elbagir. Performance of Broiler Chickens Fed Diets Containing Low Inclusion Levels of Black Cumin Seed. Journal of Animal and Veterinary Advances 2010; 9(21):2725-2728.) showed that BW and FCR were not affected in broiler fed on black cumin added ration. Al-Mufarrej (2014Al-Mufarrej SI. Immune-responsiveness and performance of broiler chickens fed black cumin (Nigella Sativa L.) powder. Journal of the Saudi Society of Agricultural Sciences 2014; 13(1): 75-80. AOAC (2000) Official Methods of Analysis. 17th Edition, The Association of Official Analytical Chemists, Gaithersburg, MD, USA.) studied the effect of black cumin on BW, BWG and FI in broilers at the level of 0.7%, 1.4%, 2.1% or 2.8%, these diets had no significant effect on BW and BWG. Güler et al. (2006) did not report a significant change in FCR of broilers containing black cumin and antibiotics. However, the rate of FCR was increased in some studies that added black cumin (Osman & El-Barody, 1999Osman AMA, El-Barody MAA. Growth performance and immune response of broiler chicks as affected by diet density and Nigella sativa seeds supplementation. Egyptian Poultry Science Journal 1999; 19: 619-634.; Al-homidan et al., 2002Al-Homidan A, Al-Qarawi AA, Al-Waily SA, Adam SEI. Response of broiler chicks to dietary Rhazya stricta and Nigella sativa. British Poultry Science 2002; 43(2): 291-296.; Abbas & Ahmed, 2010Abbas TE, Ahmed ME. Effect of supplementation of Nigella sativa seeds to the broiler chicks diet on the performance and carcass quality. International Journal of Agriculture sciences 2010; 2(2): 9.). In another study, diets containing 4% black cumin were found to consume less feed compared to the control diet and the FCR was better (Durrani et al., 2007Durrani FR, Chand N, Zaka K, Sultan A, Khattak FM, Durrani Z. Effect of Different Levels of Feed Added Black Seed (Nigella sativa L). Pakistan Journal of Biological Sciences 2007; 10(22): 4164-4167.). Saeid et al. (2013Saeid JM, Mohamed AB, AL-Baddy MA. Effect of adding garlic powder (Allium sativum) and black seed (Nigella sativa) in feed on broiler growth performance and intestinal wall structure. Journal of Natural Sciences Research 2013; 3(1): 35-41.) used the combination of %5 G and BC Premix for 42 days in commercial strain (Hubbard) broilers and found that the premix and BC positively influenced BW, BWG and FI compared to the control group. And no differences were noted in the FCR between G, BC groups and control group. These results are incompatible with the results of the current study; this incompatibilty is attributed to the strain of the broilers and differences in the duration of the study.

Some studies showed that with added BC and G there was no difference in relative organ weights (Khadr & Abdel-Fattah, 2006Khadr NA, Abdel-Fattah FAI. Response of Broiler Chickens to Diet Containing Black Seed (Nigella sativa L.) as Medical Plant. Benha Veterinary Medical Journal 2006; 17(2): 323-343.; Fallah, 2015Fallah, R. Effect of Adding Aloe vera Gel and Garlic Powder on Carcass Characteristic and Internal Organ Mass of Broiler Chickens. Global Journal of Animal Scientific Research 2015; 3 (1): 136-141.). Relative organ weights were not affected by the addition of the garlic, black cumin and G+ BC in this study too (p>0.05) (Table 3). Mahmood et al. (2009Mahmood S, Mushtaq-Ul-Hassan M, Alam M, Ahmad F. Comparative efficacy of Nigella sativa and Allium sativum as growth promoters in broilers. International Journal of Agriculture and Biology 2009; 11(6): 775-778.) and Samanthi et al. (2015) reported that the supplementation of the garlic to the ration did not affect relative organ weights (heart, stony liver and spleen). The results obtained are similar with our study.

In a study conducted by Kumar et al. (2017Kumar P, Patra AK, Mandal GP, Samanta I, Pradhan S. Effect of black cumin seeds on growth performance, nutrient utilization, immunity, gut health and nitrogen excretion in broiler chickens. Journal of the Science of Food and Agriculture, 2017; 97(11): 3742-3751.) the addition of 5, 10, 20 g/kg black cumin to the feed did not affect the morphology of the duodenum. These findings are compatible with the results of our study which showed no significant difference among trial groups in terms of the surface area of the duodenum. Lymphoid tissue is an immune system related organ that responds to tissue damage or other stimuli in the form of hyperplasia, hypertrophy, atrophy etc. On the other hand normal function of lymphoid tissue such as filtering lymph or generating antibody may also cause histomorphological changes (Haley, 2017Haley PJ. The lymphoid system: a review of species differences. Journal of toxicologic pathology 2017; 30(2): 111-123.). Molnar et al. (2011Molnár AK, Podmaniczky B, Kürti P, Tenk I, Glávits R, Virág GY, Szabó ZS. Effect of different concentrations of Bacillus subtilis on growth performance, carcase quality, gut microflora and immune response of broiler chickens, British Poultry Science 2011; 52(6): 658-665.) found increased diffuse lymphohistiocytic infiltration and solitary lymphoid follicles in the ileal mucosa, this increase was attributed to the increase in immunological responses in chickens fed with the B. subtilis supplemented diet (Molnar et al., 2011). Similarly in our study, the lymph follicles were observed to be more wide spread in the G, BC and G + BC groups than in the control. Recently, garlic has been recognised for maintaining the homeostasis of the immune system (Arreola et al., 2015Arreola R, Quintero-Fabián S, López-Roa RI, Flores-Gutiérrez EO, Reyes-Grajeda JP, Carrera-Quintanar L, Ortuño-Sahagún D. Immunomodulation and anti-inflammatory effects of garlic compounds. Journal of immunology research, 2015; 401630. https://doi.org/10.1155/2015/401630
https://doi.org/10.1155/2015/401630...
). Abdullah et al. (2019Abdullah FK, Al-Nasser AY, Al-Saffar A, Omar AE and Ragheb G. Impact of Dietary Supplementation of Different Levels of Black Seeds (Nigella sativa L.) On Production Performance, Mortality and Immunity of Broiler Chickens. International Journal of Poultry Science 2019; 18: 467-474.) supplemented black cumin seeds (0, 1 and 3%) to the diets of broilers, and found improved antibody production, as well as the immune-responsiveness of birds.

There are studies on garlic showing the inhibition of blood coagulation by platelet aggregation and platelet growth (Srivastava et al., 1993Srivastava KC, Tyagi OD: Effect of garlic-derived principle (ajoene) on aggregation and arachidonic acid metabolism in human blood platelet. Prostaglandins, Leukotrienes, and Essential Fatty Acids 1993; 49:587-595., Teranishi et al., 2003Teranishi K, Apitz-Castro R, Robson SC, Romano E, Cooper DK. Inhibition of baboon platelet aggregation in vitro and in vivo by the garlic derivative, ajoene. Xenotransplantation 2003;10(4):374-379.). Hyperemia seen in garlic group may be due to the anticoagulant effect of garlic or may be an acute immunologic response.

Measuring the biochemical parameters in a living organism plays a vital role in the diagnosis and treatment of a disease. Serum AST and ALT activity is used for the detection and differential etiologic diagnosis of hepatic diseases. The increase in the serum transaminase levels are thought to be caused by cellular release of only cytoplasmic enzymes associated with reversible hepatic cell damage (Vroon & Israili, 1990Vroon DH, Israili Z. Aminotransferases. In: Walker HK, Hall WD, Hurst JW, editors. Clinical Methods: The History, Physical, and Laboratory Examinations. 3rd edition. Boston: Butterworths; 1990. Chapter 99. Available from: https://www.ncbi.nlm.nih.gov/books/NBK425/). Creatinine is a marker that can help to evaluate the glomerular function (Gounden & Jialal, 2020Gounden V, Jialal I. Renal Function Tests. [Updated 2020 Mar 31]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing;2020 Jan-Available from: https://www.ncbi.nlm.nih.gov/books/NBK507821/). Similar to our results some researchers also found that supplementation of garlic and ginger essential oils at 10, 20 and 40 mg/kg did not cause significant effects in the activities of serum ALT or in blood creatinine level of broiler chickens (Dieumou et al. 2009Dieumou, F.E., A. Teguia, J.R. Kuiate, J.D. Tamokou, N.B. Fonge and M.C. Dongmo. Effects of ginger, (Zingiber officinale) and garlic (Allium sativum) essential oils on growth performance and gut microbial population of broiler chickens. Livestock Research for Rural Development 2009; 21(8): 23-32.). Lee et al. ( 2016Lee KW, Lee KC, Kim GH, Kim JH, Yeon JS, Cho SB, Chang BJ, Kim SK. Effects of dietary fermented garlic on the growth performance, relative organ weights, intestinal morfology, cecal microflora and serum characteristics of broiler chickens. Brazilian Journal of Poultry Science, 2016; 18 (3): 511-518.) stated that serum AST and ALT activities were decreased, but uric acid, creatinine and bilirubin values were not changed in broilers given 0.1 % fermented garlic. In accordance with our work, El-Latif et.al. (2013El-Latif ASA, Saleh NS, Allam TS, Ghazy EW. The effects of rosemary (Rosemarinus afficinalis) and garlic (Allium sativum) essential oils on performance, hematological, biochemical and immunological parameters of broiler chickens. British Journal of Poultry Sciences 2013; 2(2), 16-24.) showed that in broilers given 100 ve 200 mg/kg garlic oil the AST activity decreased, and ALT activity and uric acid value were remained unchanged. The authors indicated that garlic essential oils did not have any negative effects on liver and kidney functions (El-Latif et al. 2013).

In poultry, blood glucose level is an important physiological parameter and is affected by many factors such as diet and light regimes (Smith, 1972Smith CJV. Blood glucose levels in young chickens: The influence of light regimes. Poultry Science 1972; 51: 268-273.). El-Kaiat et al. (2002El-Kaiat AM, Soliman AZM, Hassan MSH. Combine effect of garlic, fenugreek and black seed on some productive and physiological response of laying hens. Egyptian Poultry Science 2002; 22:147-174.) found a 16% reduction in serum glucose concentration in egg chicken and Yatoo et al. (2012Yatoo MA, Sharma RK, Khan N, Rastogi A, Pathak AK. Effect of fenugreek and black cumin seeds as feed additives on blood biochemical profile and performance of broilers. Indian Journal of Animal Nutrition 2012; 29(2):174-178.) observed a significant reduction in blood glucose when using 1% black cumin in broiler diets. Similar to our results many studies have reported that blood glucose concentrations have not been affected by black cumin (Khalaji et al., 2011Khalaji S, Zaghari M, Hatami KH, Hedari-Dastjerdi S, Lotfi L, Nazarian H. Black cumin seeds, Artemisia leaves (Artemisia sieberi), and Camellia L. plant extract as phytogenic products in broiler diets and their effects on performance, blood constituents, immunity, and cecal microbial population. Poultry Science 2011; 90(11): 2500-2510., Ghasemi et al., 2014Ghasemi HA, Kasani N, Taherpour K. Effects of black cumin seed (Nigella sativa L.), a probiotic, a prebiotic and a synbiotic on growth performance, immune response and blood characteristics of male broilers. Livestock Science 2014; 164:128-134.; Kumar et al., 2017Kumar P, Patra AK, Mandal GP, Samanta I, Pradhan S. Effect of black cumin seeds on growth performance, nutrient utilization, immunity, gut health and nitrogen excretion in broiler chickens. Journal of the Science of Food and Agriculture, 2017; 97(11): 3742-3751.).

High levels of dietary cholesterol may cause atherosclerosis and coronary artery diseases in humans. Although chicken meat has low cholesterol when compared to other meat sources, changes and supplementations are made to the broiler diet to lower the level of cholesterol (Daneshyar et al., 2011Daneshyar M, Ghandkanlo MA, Sabzi Bayeghra F, Farhangpajhoh F, Aghaei M. Effects of dietary turmeric supplementation on plasma lipoproteins, meat quality and fatty acid composition in broilers. South African Journal of Animal Science 2011; 41: 420-428.). Garlic is a substance that have therapeutic effects like lowering the blood cholesterol (Lawrence & Lawrence, 2011Lawrence R, Lawrence K. Antioxidant activity of garlic essential oil (Allium sativum) grown in north Indian plains. Asian Pacific Journal of Tropical Biomedicine 2011; 1: 51-54.).

Onyimonyi et al. (2012Onyimonyi, AE, Chukwuma PC, Igbokwe C. Growth and hypocholesterolemic properties of dry garlic powder (Allium sativum) on broilers. African Journal of Biotechnology 2012; 11(11): 2666-2671.) found that the addition of 0.75% garlic to the broiler rations for 8 weeks decreased the total cholesterol level significantly when compared to the groups which added 0.25% and 0.50 garlic. Reports indicate that 1g / kg garlic powder added to broiler rations does not significantly affect serum total cholesterol concentration after a 35-day trial period (Horton, 1991Horton GMJ, Fennell MJ, Prasad BM. Effect of dietary garlic (Allium sativum) on performance, carcass composition and blood chemistry changes in broiler chickens. Canadian Journal of Animal Science 1991; 71(3): 939-942.). In a study, in which groups that received 2%, 6%, and 8% oven-dried garlic powder (2, 6 and 8) were compared with the control group, the serum total cholesterol level decreased in the groups by 19.52, 33.72 and 46.74%, respectively in the laying hens (Khan et al., 2007Khan SH, Sardar R, Anjum MA. Effects of dietary garlic on performance and serum and egg yolk cholesterol concentration in laying hens. Asian Journal of Poultry Science 2007; 1(1): 22-27.). Rahimi et al. (2011Rahimi S, Teymouri ZZ, Karimi TM, Omidbaigi R, Rokni H. Effect of the three herbal extracts on growth performance, immune system, blood factors and intestinal selected bacterial population in broiler chickens. Journal of Agricultural Science and Technology 2011;13: 527-539.) added 0.1% garlic powder to broiler ration, and showed that the HDL-cholesterol level increased while total cholesterol level decreased. Issa and Omar (2012Issa KJ, Omar JMA. Effects of garlic powder on performance and lipid profile of broilers. Open Journal of Animal Science 2012; 2: 62-68.) showed that in the groups that received 0.2% and 0.4% garlic powder, the blood total cholesterol level decreased, while HDL-cholesterol level increased in Cobb broilers compared to the control. Garlic thought to reduce the total cholesterol by inhibiting the synthesis of harmful LDL cholestrol and by increasing beneficial HDL cholesterol in the blood. The mechanism of the hypocholesterolemic and hypolipidemic effects of garlic products is believed to be effective by depressing the cholestogenic and lipogenic activities of liver enzymes such as malic enzyme, fatty acid synthase, glucose 6 phosphate dehydrogenase and 3-hydroxy-3-methyl glutalyl CoA reductase (Qureshi et al., 1983Qureshi AA, Abuirmeileh N, Din ZZ, Elson CE, Burger WC. Inhibition of cholesterol and fatty acid biosynthesis in liver enzymes and chicken hepatocytes by polar fractions of garlic. Lipids 1983; 18(5): 343-348.; Mahmoud et al., 2010Mahmoud KZ, Gharaibeh SM, Zakaria HA, Qatramiz AM. Garlic (Allium sativum) supplementation: Influence on egg production, quality, and yolk cholesterol level in layer hens. Asian-Australasian Journal of Animal Sciences 2010; 23(11):1503-1509.). In this study no significant change was detected in the level of total cholesterol and HDL cholesterol among trial groups.

Blood proteins in birds is an important indicator while estimating the health of the animal and helps to evaluate the biochemical metabolic changes. The changes in the blood protein levels generally depends on the alterations in the diet (Tóthová et al., 2019Tóthová C, Sesztáková E, Bielik B, Nagy O. Changes of total protein and protein fractions in broiler chickens during the fattening period. Veterinary World 2019; 12(4), 598-604.). In a study conducted by Kumar et al. (2017Kumar P, Patra AK, Mandal GP, Samanta I, Pradhan S. Effect of black cumin seeds on growth performance, nutrient utilization, immunity, gut health and nitrogen excretion in broiler chickens. Journal of the Science of Food and Agriculture, 2017; 97(11): 3742-3751.) the total protein concentration tended to be higher in the groups fed on black cumin than the control groups. The same results had been found by Hassan et al. (2007Hassan MSH, AboTaleb AM, Wakwak MM, Yousef BA. Productive physiological and immunological effect of using some natural feed additives in Japanese quail. Egyptian Poultry Science 2007; 27: 557-581.) who showed high levels of serum total protein, albumin and globulin in broilers fed on high levels of black cumin. Total serum protein increased with the addition of black cumin (Al-Beitawi et al., 2009Al-Beitawi NA, El-Ghousein SS, Nofal AH. Replacing bacitracin methylene disalicylate by crushed Nigella sativa seeds in broiler rations and its effects on growth, blood constituents and immunity. Livestock Science 2009;125(2-3): 304-307.; Khan et al., 2012Khan RU, Nikousefat Z, Tufarelli V, Naz S, Javdani M, Laudadio V. Garlic (Allium sativum) supplementation in poultry diets: effects on production and physiology. World' s Poultry Science Journal 2012; 68(3):417-424.; Yatoo et al., 2012Yatoo MA, Sharma RK, Khan N, Rastogi A, Pathak AK. Effect of fenugreek and black cumin seeds as feed additives on blood biochemical profile and performance of broilers. Indian Journal of Animal Nutrition 2012; 29(2):174-178.; Saleh 2014Saleh AA. Nigella seed oil as alternative to avilamycin antibiotic in broiler chicken diets. South African Journal of Animal Science 2014; 44(3): 254-261.). However, El-Ghammry et al. (2002El-Ghammry AA, El-Mallah GM El-Yamny AT. The effect of incorporation yeast culture, Nigella sativa seeds and fresh garlic in broiler diets on their performance. Egyptian Poultry Science 2002; 22: 445-459.) and Toghyani et al. (2010Toghyani MA, Geisari G, Ghalamkari M, Mohammadrezaei M. Growth performance, serum biochemistry and blood hematology of broiler chicks fed different levels of black seed (Nigella sativa L.) and peppermint (Mentha piperita). Livestock Science 2010; 129: 173-178.) found that black cumin does not affect plasma total protein, albumin and globulin concentrations. Hermes et al. (2011Hermes H, Attia FM, Ibrahim KA, El-Nesr SS. Physiological responses of broiler chickens to dietary different forms and levels of Nigella sativa L. during Egyptian summer season. Journal of Agricultural and Veterinary Science 2011; 4:17-33.) observed an increase in total plasma proteins as well as albumin and globulin in black cumin group as compared to the control group, but the differences were not significant. The recorded increase of albumin (p<0.001) and total protein (p<0.05) in BC and G+BC groups of this study can be attributed to the BC as it contains more protein and its immuno stimulating effect.

In recent years many scientist showed interest in researches about oxidative stress and antioxidant agents. Among these antioxidant agents natural substances attracted most of the attention because these substances can prevent diseases and positively affect the health (Hassan et al., 2018Hassan F, Roushdy EM, Kishawy, Zaglool, AW, Tukur HA, Saadeldin IM. Growth Performance, Antioxidant Capacity, Lipid-Related Transcript Expression and the Economics of Broiler Chickens Fed Different Levels of Rutin. Animals: an open access journal from MDPI, 2018; 9(1): 7.). The efficacy of garlic preparations or extracts to alleviate free radical damage to biological membranes or other biological systems has been demonstrated (Horie et al., 1989Horie T, Murayama T, Mishima T, Itoh F, Minamide Y, Fuwa T, Awazu S. Protection of liver microsomal membranes from lipid peroxidation by garlic extract. Journal of Medicinal Plant and Natural 1989; 55:506-508.; Knasmüller et al., 1989Knasmüller S, De Martin R, Domjan G, Szakmary A. Studies on the antimutagenic activities of garlic extract. Environmental and Molecular Mutagenesis 1989; 13:357-365.; Kourounakis & Rekka, 1991Kourounakis PN, Rekka EA. Effect on active oxygen species of alliin and Allium sativum (garlic) powder. Research Communications in Chemical Pathology and Pharmacology Journal 1991; 74: 249-252.; Chung, 2006Chung LY. The Antioxidant Properties of Garlic Compounds: Allyl Cysteine, Alliin, Allicin, and Allyl Disulfide. Journal of Medicinal Food 2006; 9 (2): 205-213.). Pourali et al. (2014Pourali M, Kermanshahi H, Golia A., Razmi G, Soukhtanloo M. Antioxidant and anticoccidial effects of garlic powder and sulfur amino acids on Eimeria-infected and uninfected broiler chickens. Iranian Journal of Veterinary Research 2014; 15(3), 227-232.) studied the antioxidant effect of 0.5 % garlic powder on Eimeria infected and uninfected broiler chickens; and found that the MDA levels were decreased in 0.5% garlic powder group as compared to the not given group in infected broilers showing the antioxidant effect of the garlic powder. On the other hand, similar to our study no statistically significant difference was recorded in the TOS between control and 0.5% garlic powder group in the uninfected broilers. Pourali et al. (2014) showed no difference in the SOD and GPX activities of control and 0.5% garlic powder received group in uninfected broiler chickens. This result is compatible with our study as no difference was found in TAS between 0.5% G and control group. Black cumin is an excellent superoxide anion scavenger that prevents oxidative damage (Azeema et al., 2014Azeema T, Rehmanb ZU, Umar S, Asifa M, Arif M, Rahman A. Effect of Nigella Sativa on poultry health and production: A review. Science Letters 2014; 2: 76-82.). Tuluce et al. (2009Tuluce YH, Ozkol B, Sogut I, Celik. Effects of Nigella sativa on lipid peroxidation and reduced glutathione levels in erythrocytes of broiler chickens. Cell Membranes and Free Radical Research 2009;1:1-3.) added 0.5 %, 1 % and 1.5 % BC to the diet of broilers for 6 weeks, and found that erythrocyte MDA levels were significantly decreased in 0.5 % and 1 % BC groups. Guler et al. (2007Guler T, Ertas ON, Kizil M, Dalkilic B, Ciftci M. Effect of dietary supplemental black cumin seeds on antioxidant activity in broilers. Medycyna Weterynaryjna 2007; 63: 1060-1063.) studied black cumin seeds at 0.5%, 1%, 2% or 3% in the basal diet of broilers for 42 days and found that 2% and 3% BC seed reduced the MDA concentration in serum when compared to birds fed 1% and 0.5% black cumin seeds and control diet. These results showed that the antioxidant effect of BC depends on the dose supplemented to the diet.

As a result, although not statistically significant, numerical improvement in FCR was observed in groups given garlic and garlic + black cumin. In addition, there are no differences in TAS, TOS levels. In histological examinations hyperemia was observed, especially in the G group, in overlapping macroscopic findings in lamina propria and other viticular areas. The lymph follicles were observed to be more wide spread in the G, BC and G+BC groups as compared to the control. The results of the study showed that the combination of BC and G can have beneficial effects, and doses of G and BC may be the reason for the positive effects as compared to no effect.

ACKNOWLEDGEMENT

This research was supported by the Scientific Research Fund in the University of Kirikkale (Project No: 2017/065).

REFERENCES

  • Abbas TE, Ahmed ME. Effect of supplementation of Nigella sativa seeds to the broiler chicks diet on the performance and carcass quality. International Journal of Agriculture sciences 2010; 2(2): 9.
  • Abdullah FK, Al-Nasser AY, Al-Saffar A, Omar AE and Ragheb G. Impact of Dietary Supplementation of Different Levels of Black Seeds (Nigella sativa L.) On Production Performance, Mortality and Immunity of Broiler Chickens. International Journal of Poultry Science 2019; 18: 467-474.
  • Adebiyi FG, Ologhobo AD, Adejumo IO. Efficacy of Allium sativum as growth promoter, immune booster and cholesterollowering agent on broiler chickens. Asian Journal of Animal Science 2017; 11(5): 202-213.
  • Al-Beitawi NA, El-Ghousein SS, Nofal AH. Replacing bacitracin methylene disalicylate by crushed Nigella sativa seeds in broiler rations and its effects on growth, blood constituents and immunity. Livestock Science 2009;125(2-3): 304-307.
  • Al-Homidan A, Al-Qarawi AA, Al-Waily SA, Adam SEI. Response of broiler chicks to dietary Rhazya stricta and Nigella sativa. British Poultry Science 2002; 43(2): 291-296.
  • Al-Mufarrej SI. Immune-responsiveness and performance of broiler chickens fed black cumin (Nigella Sativa L.) powder. Journal of the Saudi Society of Agricultural Sciences 2014; 13(1): 75-80. AOAC (2000) Official Methods of Analysis. 17th Edition, The Association of Official Analytical Chemists, Gaithersburg, MD, USA.
  • Apata DF. Antibiotic resistance in poultry. International Journal of Poultry Science 2009; 8(4), 404-408.
  • Arreola R, Quintero-Fabián S, López-Roa RI, Flores-Gutiérrez EO, Reyes-Grajeda JP, Carrera-Quintanar L, Ortuño-Sahagún D. Immunomodulation and anti-inflammatory effects of garlic compounds. Journal of immunology research, 2015; 401630. https://doi.org/10.1155/2015/401630
    » https://doi.org/10.1155/2015/401630
  • Ashayerizadeh O, Dastar B, Shargh MS. Use of garlic (Allium sativum), black cumin seeds (Nigella sativa L.) and wild mint (Mentha longifolia) in broiler chicken diets. Journal of Animal and Veterinary Advances 2009; 8(9): 1860-1867.
  • Azeema T, Rehmanb ZU, Umar S, Asifa M, Arif M, Rahman A. Effect of Nigella Sativa on poultry health and production: A review. Science Letters 2014; 2: 76-82.
  • Borgohain B, Mahanta JD, Islam R, Sapcota D, Sarma S, Borah MC. Effect of feeding garlic (Allium sativum) as prebiotic on the performance of broiler chicken. International Journal of Livestock Research 2017; 7(7): 225-233.
  • Chung LY. The Antioxidant Properties of Garlic Compounds: Allyl Cysteine, Alliin, Allicin, and Allyl Disulfide. Journal of Medicinal Food 2006; 9 (2): 205-213.
  • Daneshyar M, Ghandkanlo MA, Sabzi Bayeghra F, Farhangpajhoh F, Aghaei M. Effects of dietary turmeric supplementation on plasma lipoproteins, meat quality and fatty acid composition in broilers. South African Journal of Animal Science 2011; 41: 420-428.
  • Denk W, Webb WW, Hudspeth AJ. Mechanical properties of sensory hair bundles are reflected in their Brownian motion measured with a laser differential interferometer. Proceedings of the National Academy of Sciences 1989; 86(14): 5371-5375.
  • Dieumou, F.E., A. Teguia, J.R. Kuiate, J.D. Tamokou, N.B. Fonge and M.C. Dongmo. Effects of ginger, (Zingiber officinale) and garlic (Allium sativum) essential oils on growth performance and gut microbial population of broiler chickens. Livestock Research for Rural Development 2009; 21(8): 23-32.
  • Durrani FR, Chand N, Zaka K, Sultan A, Khattak FM, Durrani Z. Effect of Different Levels of Feed Added Black Seed (Nigella sativa L). Pakistan Journal of Biological Sciences 2007; 10(22): 4164-4167.
  • Elagib HAA, El-Amin WIA, Elamin KM, Malik HEE. Effect of dietary garlic (Allium sativum) supplementation as feed additive on broiler performance and blood profile. Journal of Animal science advances 2013; 3(2): 58-64.
  • El-Ghammry AA, El-Mallah GM El-Yamny AT. The effect of incorporation yeast culture, Nigella sativa seeds and fresh garlic in broiler diets on their performance. Egyptian Poultry Science 2002; 22: 445-459.
  • El-Kaiat AM, Soliman AZM, Hassan MSH. Combine effect of garlic, fenugreek and black seed on some productive and physiological response of laying hens. Egyptian Poultry Science 2002; 22:147-174.
  • El-Latif ASA, Saleh NS, Allam TS, Ghazy EW. The effects of rosemary (Rosemarinus afficinalis) and garlic (Allium sativum) essential oils on performance, hematological, biochemical and immunological parameters of broiler chickens. British Journal of Poultry Sciences 2013; 2(2), 16-24.
  • Erel O. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clinical Biochemistry 2004; 37(4), 277-285.
  • Erel O. A new automated colorimetric method for measuring total oxidant status. Clinical biochemistry 2005; 38(12):1103-1111.
  • Fallah, R. Effect of Adding Aloe vera Gel and Garlic Powder on Carcass Characteristic and Internal Organ Mass of Broiler Chickens. Global Journal of Animal Scientific Research 2015; 3 (1): 136-141.
  • Ghasemi HA, Kasani N, Taherpour K. Effects of black cumin seed (Nigella sativa L.), a probiotic, a prebiotic and a synbiotic on growth performance, immune response and blood characteristics of male broilers. Livestock Science 2014; 164:128-134.
  • Gounden V, Jialal I. Renal Function Tests. [Updated 2020 Mar 31]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing;2020 Jan-Available from: https://www.ncbi.nlm.nih.gov/books/NBK507821/
  • Guler T, Ertas ON. The effect of dietary black cumin seeds (Nigella sativa L.) on the performance of broilers. Asian-Australasian Journal of Animal Sciences 2006; 19(3): 425-430.
  • Guler T, Ertas ON, Kizil M, Dalkilic B, Ciftci M. Effect of dietary supplemental black cumin seeds on antioxidant activity in broilers. Medycyna Weterynaryjna 2007; 63: 1060-1063.
  • Gundersen HJG, Jensen EB. The efficiency of systematic sampling in stereology and its prediction. Journal of microscopy 1987; 147(3): 229-263.
  • Guo F. Mushroom and herb polysachariides as alternative for antimicrobial growth promotors in poultry. 2003.
  • Haley PJ. The lymphoid system: a review of species differences. Journal of toxicologic pathology 2017; 30(2): 111-123.
  • Hassan MSH, AboTaleb AM, Wakwak MM, Yousef BA. Productive physiological and immunological effect of using some natural feed additives in Japanese quail. Egyptian Poultry Science 2007; 27: 557-581.
  • Hassan F, Roushdy EM, Kishawy, Zaglool, AW, Tukur HA, Saadeldin IM. Growth Performance, Antioxidant Capacity, Lipid-Related Transcript Expression and the Economics of Broiler Chickens Fed Different Levels of Rutin. Animals: an open access journal from MDPI, 2018; 9(1): 7.
  • Hermes H, Attia FM, Ibrahim KA, El-Nesr SS. Physiological responses of broiler chickens to dietary different forms and levels of Nigella sativa L. during Egyptian summer season. Journal of Agricultural and Veterinary Science 2011; 4:17-33.
  • Horie T, Murayama T, Mishima T, Itoh F, Minamide Y, Fuwa T, Awazu S. Protection of liver microsomal membranes from lipid peroxidation by garlic extract. Journal of Medicinal Plant and Natural 1989; 55:506-508.
  • Horton GMJ, Fennell MJ, Prasad BM. Effect of dietary garlic (Allium sativum) on performance, carcass composition and blood chemistry changes in broiler chickens. Canadian Journal of Animal Science 1991; 71(3): 939-942.
  • Issa KJ, Omar JMA. Effects of garlic powder on performance and lipid profile of broilers. Open Journal of Animal Science 2012; 2: 62-68.
  • Kansal A, Ali N, Fahim A, Bharti M, Chandra G, Siddiqui R. Effect of Dietary Supplementation of Garlic on the Haemato-biochemical Parameters and Performance of Broiler Chickens. International Journal of Livestock Research 2017; 7(10): 223-230.
  • Khadr NA, Abdel-Fattah FAI. Response of Broiler Chickens to Diet Containing Black Seed (Nigella sativa L.) as Medical Plant. Benha Veterinary Medical Journal 2006; 17(2): 323-343.
  • Khalaji S, Zaghari M, Hatami KH, Hedari-Dastjerdi S, Lotfi L, Nazarian H. Black cumin seeds, Artemisia leaves (Artemisia sieberi), and Camellia L. plant extract as phytogenic products in broiler diets and their effects on performance, blood constituents, immunity, and cecal microbial population. Poultry Science 2011; 90(11): 2500-2510.
  • Khan SH, Sardar R, Anjum MA. Effects of dietary garlic on performance and serum and egg yolk cholesterol concentration in laying hens. Asian Journal of Poultry Science 2007; 1(1): 22-27.
  • Khan RU, Nikousefat Z, Tufarelli V, Naz S, Javdani M, Laudadio V. Garlic (Allium sativum) supplementation in poultry diets: effects on production and physiology. World' s Poultry Science Journal 2012; 68(3):417-424.
  • Knasmüller S, De Martin R, Domjan G, Szakmary A. Studies on the antimutagenic activities of garlic extract. Environmental and Molecular Mutagenesis 1989; 13:357-365.
  • Kodera Y, Ushijima M, Amano H, Suzuki JI, Matsutomo T. Chemical and Biological Properties of S-1-Propenyl-L-Cysteine in Aged Garlic Extract. Molecules, 2017; 22(4):570.
  • Konjufca VH, Pesti GM, Bakalli RI. Modulation of cholesterol levels in broiler meat by dietary garlic and copper. Poultry science 1997; 76(9):1264-1271.
  • Kourounakis PN, Rekka EA. Effect on active oxygen species of alliin and Allium sativum (garlic) powder. Research Communications in Chemical Pathology and Pharmacology Journal 1991; 74: 249-252.
  • Kubínová L, Janacek J. Estimating surface area by the isotropic Fakir method from thick slices cut in an arbitrary direction. Journal of Microscopy 1998; 191(2):201-211.
  • Kumar P, Patra AK, Mandal GP, Samanta I, Pradhan S. Effect of black cumin seeds on growth performance, nutrient utilization, immunity, gut health and nitrogen excretion in broiler chickens. Journal of the Science of Food and Agriculture, 2017; 97(11): 3742-3751.
  • Lawrence R, Lawrence K. Antioxidant activity of garlic essential oil (Allium sativum) grown in north Indian plains. Asian Pacific Journal of Tropical Biomedicine 2011; 1: 51-54.
  • Lee KW, Lee KC, Kim GH, Kim JH, Yeon JS, Cho SB, Chang BJ, Kim SK. Effects of dietary fermented garlic on the growth performance, relative organ weights, intestinal morfology, cecal microflora and serum characteristics of broiler chickens. Brazilian Journal of Poultry Science, 2016; 18 (3): 511-518.
  • Lewis MN, Rose SP, Mackenzie AM, Tucker LA. Effects of dietary inclusion of plant extracts on the growth performance of male broiler chickens. Spring Meetingof the WPSA UK Branch Posters 2003; 43-44.
  • Lymia HA, MajeedKhadiga A, Abdelati Nabiela M Elbagir. Performance of Broiler Chickens Fed Diets Containing Low Inclusion Levels of Black Cumin Seed. Journal of Animal and Veterinary Advances 2010; 9(21):2725-2728.
  • Mahmood S, Mushtaq-Ul-Hassan M, Alam M, Ahmad F. Comparative efficacy of Nigella sativa and Allium sativum as growth promoters in broilers. International Journal of Agriculture and Biology 2009; 11(6): 775-778.
  • Mahmoud KZ, Gharaibeh SM, Zakaria HA, Qatramiz AM. Garlic (Allium sativum) supplementation: Influence on egg production, quality, and yolk cholesterol level in layer hens. Asian-Australasian Journal of Animal Sciences 2010; 23(11):1503-1509.
  • Mansoub NH, Mohannad AMN. Effect of garlic, thyme and yogurt compared to antibiotic on performance, immunity and some blood parameters of broiler chicken. Indian Journal of Animal Science 2011; 81(12): 1197-1200.
  • Molnár AK, Podmaniczky B, Kürti P, Tenk I, Glávits R, Virág GY, Szabó ZS. Effect of different concentrations of Bacillus subtilis on growth performance, carcase quality, gut microflora and immune response of broiler chickens, British Poultry Science 2011; 52(6): 658-665.
  • Onyimonyi, AE, Chukwuma PC, Igbokwe C. Growth and hypocholesterolemic properties of dry garlic powder (Allium sativum) on broilers. African Journal of Biotechnology 2012; 11(11): 2666-2671.
  • Osman AMA, El-Barody MAA. Growth performance and immune response of broiler chicks as affected by diet density and Nigella sativa seeds supplementation. Egyptian Poultry Science Journal 1999; 19: 619-634.
  • Pourali M, Kermanshahi H, Golia A., Razmi G, Soukhtanloo M. Antioxidant and anticoccidial effects of garlic powder and sulfur amino acids on Eimeria-infected and uninfected broiler chickens. Iranian Journal of Veterinary Research 2014; 15(3), 227-232.
  • Qureshi AA, Abuirmeileh N, Din ZZ, Elson CE, Burger WC. Inhibition of cholesterol and fatty acid biosynthesis in liver enzymes and chicken hepatocytes by polar fractions of garlic. Lipids 1983; 18(5): 343-348.
  • Rahimi S, Teymouri ZZ, Karimi TM, Omidbaigi R, Rokni H. Effect of the three herbal extracts on growth performance, immune system, blood factors and intestinal selected bacterial population in broiler chickens. Journal of Agricultural Science and Technology 2011;13: 527-539.
  • Rehman Z, Munir MT. Effect of garlic on the health and performance of broilers. Veterinaria 2015; 3(1): 32-39.
  • Rozbeh F. Effect of adding aloe vera gel and garlic powder on carcass characteristic and internal organ mass of broiler chickens. Global Journal of Animal Scientific Research 2015; 3(1):136-141.
  • Saeid JM, Mohamed AB, AL-Baddy MA. Effect of adding garlic powder (Allium sativum) and black seed (Nigella sativa) in feed on broiler growth performance and intestinal wall structure. Journal of Natural Sciences Research 2013; 3(1): 35-41.
  • Saleh AA. Nigella seed oil as alternative to avilamycin antibiotic in broiler chicken diets. South African Journal of Animal Science 2014; 44(3): 254-261.
  • Samanthi KAM, Nayananjalie WAD, Adikari AMJR, Liyanage R. Dietary garlic (Allium sativum L.) supplementation on performance, meat quality and lipid profile in broilers. Annual Symposium. 2014, 6.
  • Singh and Kumar. Effect of dietary black cumin (Nigella sativa) on the growth performance, nutrient utilization, blood biochemical profile and carcass traits in broiler chickens. Animal Nutrition and Feed Technology 2018; 18:409-419.
  • Smith CJV. Blood glucose levels in young chickens: The influence of light regimes. Poultry Science 1972; 51: 268-273.
  • Soliman AZM, Ghazalah AA, El-Samra SH, Atta AM, Abdo ZMA. The synergistic effect of either black seeds or garlic with fat on broiler performance and immunity. Egyptian Journal of Nutrition and Feeds 1999; 2: 603-620.
  • Srivastava KC, Tyagi OD: Effect of garlic-derived principle (ajoene) on aggregation and arachidonic acid metabolism in human blood platelet. Prostaglandins, Leukotrienes, and Essential Fatty Acids 1993; 49:587-595.
  • Teranishi K, Apitz-Castro R, Robson SC, Romano E, Cooper DK. Inhibition of baboon platelet aggregation in vitro and in vivo by the garlic derivative, ajoene. Xenotransplantation 2003;10(4):374-379.
  • Toghyani MA, Geisari G, Ghalamkari M, Mohammadrezaei M. Growth performance, serum biochemistry and blood hematology of broiler chicks fed different levels of black seed (Nigella sativa L.) and peppermint (Mentha piperita). Livestock Science 2010; 129: 173-178.
  • Tollba AH, Hassan MH. Using some natural additives to improve physiological and productive performance of broiler chicks under high temperature conditions. 2-Black cumin (Nigella sativa) or garlic (Allium sativum). Egyptian Poultry Science 2003; 23: 327-340.
  • Tóthová C, Sesztáková E, Bielik B, Nagy O. Changes of total protein and protein fractions in broiler chickens during the fattening period. Veterinary World 2019; 12(4), 598-604.
  • Tuluce YH, Ozkol B, Sogut I, Celik. Effects of Nigella sativa on lipid peroxidation and reduced glutathione levels in erythrocytes of broiler chickens. Cell Membranes and Free Radical Research 2009;1:1-3.
  • Vroon DH, Israili Z. Aminotransferases. In: Walker HK, Hall WD, Hurst JW, editors. Clinical Methods: The History, Physical, and Laboratory Examinations. 3rd edition. Boston: Butterworths; 1990. Chapter 99. Available from: https://www.ncbi.nlm.nih.gov/books/NBK425/
  • Wang R, Li D, Bourne S. Can 2000 years of herbal medicine history help us solve problems in the year 2000. In Alltechs annual symposium 1998; Vol. 14, pp. 168-184.
  • Yatoo MA, Sharma RK, Khan N, Rastogi A, Pathak AK. Effect of fenugreek and black cumin seeds as feed additives on blood biochemical profile and performance of broilers. Indian Journal of Animal Nutrition 2012; 29(2):174-178.
  • Yildirim E, Çinar M. Herbal treatment approaches in metal poisoning. Turkiye Klinikleri Journal Vet Sci Pharmacology Toxicology-Special Topics 2017; 3(1):50-6.
  • Yoo M, Lee S, Kim S, Hwang JB, Choe J, Shin D. Composition of organosulfur compounds from cool-and warm-type garlic (Allium sativum L.) in Korea. Food Science and Biotechnology 2014; 23(2):337-344.
  • Zahoor A, Ghafoor A, Aslam M. Nigella sativa-a potential commodity in crop diversification traditionally used in healthcare. Project on introduction of medicinal herbs and spices as crops. Ministry of Food. Agriculture and Livestock Education 2004; 280-295.

Publication Dates

  • Publication in this collection
    01 Dec 2020
  • Date of issue
    2020

History

  • Received
    07 May 2020
  • Accepted
    05 Oct 2020
Fundação APINCO de Ciência e Tecnologia Avícolas Av. Andrade Neves, 2501 - Castelo, 13070-001 Campinas SP - Brazil, Tel.: (55 19) 3243-6555 / Fax.: (55 19) 3243-8542 - Campinas - SP - Brazil
E-mail: rvfacta@terra.com.br