Effect of dietary BHA supplementation on certain physiological values in broiler chicken

Hadeel M. Hameed¹, Hiyam N. Maty², Ashwaq A. Hassan³

Department of Physiology, Biochemistry, and Pharmacology, College of Veterinary Medicine, University of Mosul, Iraq.

hadeel.mohammad@uomosul.edu.iq, 0000-0002-4858-2391

hemyatem@yahoo.com, 0000-0003-1792-5192

d.ashwaqah@yahoo.com. 0000-0003-4883-6192

Abstract

Butylated hydroxyl anisole (BHA) is manufactured from a mixture of two isomeric organic compounds, 2-tertiary-butyl-4-hydroxyanisole and 3-tert-butyl-4-hydroxyanisole, and is widely used in poultry feed as a phenolic free radical scavenger. The current search investigated the impact of different doses of BHA on some hematological and hormonal features of broiler. Fifty-four one-day-old birds were utilized in this study randomly divided into three groups, the control group was given a standard ratio, the 2^ed group was given a standard diet filled up with BHA at a rate of 2.5 mg/kg feed, and the 3^rd group was given a standard ration fill up with BHA at a rate of 5 mg/kg feed by 18 birds/group, each with three replicates (6 birds/ repeated cycle). The study included three age stages (15, 30, and 45) days. The results showed a significant rise in RBC, WBC, lymphocytes, MCH, SOD, and CAT for the two groups of BHA during 30 and 45 days of treatment compared with the control with a significant improvement in the stress index, as well as a significant decrease in MCV during the three periods of treatment. Supplementation with BHA in both doses caused a significant rise in T3 and T4 during 45 days of the study compared with the control group. We deduce from this research that adding BHA to the poultry diets improved physiological and hormonal characteristics due to its ability to scavenge free radicals. It promotes significant health and growth.

Keywords: BHA, Hematology, T3, T4, Broiler

التأثیر الغذائی لإضافة البوتیل هیدروکسی الأنیسول فی بعض القیم الفسلجیة لأفراخ فروج اللحم

هدیل محمد حمید1، هیام نذیر متی2 و اشواق احمد حسن3

فرع الفسلجة والکیمیاء الحیاتیة والأدویة، کلیة الطب البیطری، جامعة الموصل، الموصل، العراق

الخلاصة

یعتبر البوتیل هیدروکسی الأنیسول من مضادات الاکسدة المصنعة من خلیط من مرکبین عضویین ایزومیریین، 2-ثالثی-بیوتیل-4-هیدروکسی انیسول و 3-ثلاثی-بیوتیل-4-هیدروکسی انیسول یستخدم بشکل واسع النطاق فی اعلاف الدواجن کمواد فینولیة کاسحة للجذور الحرة. تناولت الدراسة الحالیة تاثیر مستویات مختلفة من البوتیل هیدروکسی الأنیسول فی بعض الصفات الدمویة والهرمونیة لفروج اللحم. استخدم فی هذه الدراسة 54 طائر بعمر یوم قسمت مجامیع الدراسة بشکل عشوائی الى ثلاث مجامیع وهی مجموعة السیطرة اعطیت علیقة قیاسیة والمجموعة الثانیة اعطیت علیقة قیاسیة مضاف الیها بوتیل هیدروکسی الأنیسول بترکیز 2,5 ملغم/کغم علف والمجموعة الثالثة اعطیت علیقة قیاسیة مضاف الیها البوتیل هیدروکسی الأنیسول بترکیز5ملغم/کغم علف وبواقع 18 طائر/ لکل مجموعة وقسمت کل مجموعة الى ثلاث مکررات بواقع 6طائر/ مکرر وشملت الدراسة ثلاث مراحل عمریة عند 15، 30 و 45 یوم. اظهرت النتائج زیادة معنویة فی خلایا الدم البیض، خلایا الدم الحمر,الخلایا اللمفاویة,معدل هیموکلوبین الکریة,سوبر اوکسید دسمیوتیز وانزیم الکاتالیز مع تحسن معنوی فی دلیل الکرب اضافة الى انخفاض معنوی فی معدل حجم الکریة خلال الفترات الثلاثة من المعاملة. تجهیز البوتیل هیدروکسی الأنیسول بجرعتیه ادى الى ارتفاع معنوی فی مستوى هرمون الثایروکسین وثلاثی ایوداید الثایرونین خلال فترة 45 یوم من الدراسة مقارنة مع السیطرة. استنتج من هذه الدراسة ان اضافة البوتیل هیدروکسی الأنیسول فی غذاء الدواجن یحسن من بعض الصفات الدمویة والهرمونیة نتیجة لدوره الکاسح للجذور الحرة مما یؤدی الى صحة ونمو أفضل.

Introduction

During previous years, different types of food additives were used in poultry rations, including probiotics, organic acids, and others, as (1) indicated that adding the probiotic and emulsifiers to quail diets at a rate of 0.5 mg/kg of feed led to an increase in the level of the FSH and LH, as indicated by (2) that adding organic acids to quail diet led to an elevated in the level of growth hormone. BHA is one of the additives used in poultry diets, but the references to study the physiological importance of BHA. BHA is an industrial antioxidant widely used in animal diets as a free radical scavenger (3). Synthetic antioxidants are manufactured in different ways, and in general, they are phenolic compounds that mainly work to capture free radicals and stop oxidative reactions. Industrial antioxidants are used in various fields, significantly as feed additives in poultry diets, to improve growth and production. BHA is a fat-soluble antioxidant (4), and it is a phenolic preservative similar in composition to vitamin E and is widely used in many industries and feed additives (5,6). It has a vital role in inhibiting the oxidation process of fats by increasing the effectiveness of antioxidants. It was also recorded previously has a remarkable function effect in protecting the liver from oxidative stress when added at a concentration of 0.5 mg/kg to poultry diets (5).

Materials and methods

Animals study

This study was conducted in the College of Veterinary Medicine, Mosul. In this study, 54 rose-bred birds were used at the age of one day until the end of the experiment to 45 days under appropriate conditions of temperature and light. The birds were fed with two rations, the growth and finisher ration. Ration and water were available freely, according to NRC (7).

Experimental design

Fifty-four birds were utilized in this experiment, randomly partitioned into three groups. Each group included 18 birds/group, and each group was divided into three replicates, six birds/repeat. The birds were divided into three age stages (15, 30, and 45). The study groups included the following; the control group was given a standard diet without the addition. In the second group, a standard diet was given to which BHA was added at a rate of 2.5 mg/kg of feed. In the third group, a standard diet was given to which BHA was added at a rate of 5 mg/kg of feed.

Studied traits

After the end of the experiment, the birds were slaughtered by cutting the jugular vein. The blood samples were collected and divided into two parts. The first part was placed in test tubes containing anticoagulants (heparinized tubes) to conduct blood tests. The blood tests checked for red blood cell count (RBC), white blood cell count (WBC), leukocyte differential count (DLC), stress index (heterophils/lymphocytes), hemoglobin concertation (Hb), packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concertation (MCHC). According to Campbell (8), the other part was placed in tubes that did not contain anticoagulants and left to clot at room temperature. It was centrifuged at a velocity of 3000 rpm for 15 minutes to impose the serum were kept at -20°C to estimate the level of thyroid hormones using ELISA technique (9), catalase, and superoxide dismutase by using commercial kits.

Statistical analysis

Data was determined using the rate of a one-way analysis of variance (ANOVA). The significance of variations among means was assessed and used the Duncan Multiple Range Test at P

Results

The results shown in table 1 in the current study indicate a significant rise in the number of WBC in the BHA group supplied with a dose of 5 mg during the 30-45 days of the study. The BHA group with a dose of 2.5 mg during 45 days compared with the control. Treatment with both groups of BHA during the 30-45 days of treatment indicates a significant elevate in the number of lymphocytes compared with the control. There was a significant decrease in the number of heterotrophils of BHA groups during the 45 days of the study compared with the other group. Treatment of the BHA and the control group at a dose of 5 mg during the 45th period of the study and the BHA at the dose of 2.5 mg during the 30 days of treatment decreased the rate of basophils in comparison with the control and the BHA at a dose of 5 mg during the 30 days of the study. As for the results, the number of eosinophil treatments with the BHA and control groups during the periods 30 and 45 days led to a significant rise in their numbers compared to the control during 15 days of the experiment. While results of the BHA and control showed during 30 and 45 days of the experiment indicate a significant improvement in the stress index compared with the control during 15 days of treatment.

Table 1: Effect of BHA on white blood cell count and DLC in broiler after 15, 30, and 45 days of treatments

Values using various crafts in the pillar are significantly diverse at P<0.05.

The data in Table 2 shows a significant elevate in the number of RBC for the BHA groups during 45 days of the study compared to the control. The treatment with BHA at a dose of 5 mg in the 30 and 45 days of the study and the BHA group at a dose of 2.5 mg within 45 days to a significant rise in hemoglobin level compared with the control group. The two BHA groups caused a significant elevation in PCV levels during 30 and 45 days compared to the control.

Table 2: Effect of BHA on blood picture in broiler after 15, 30, and 45 days of treatments

Values using various crafts in the pillar are significantly diverse at P<0.05.

Table 3 referred to a significant reduction in the rate of MCV for the two BHA groups during 15, 30, and 45 days of the study compared with the control. Treatment with BHA at a dose of 5mg led to a significant increase in the level of MCH during 30 days of the treatment compared with the control and BHA groups at a dose of 2.5 mg, while treatment with BHA at a dose of 5 mg caused a significant rise in the level of MCHC during 30 and 45 days of the study, and the BHA group at a dose of 2.5 mg during 45 days compared with the control.

Table 3: Effect of BHA on some blood parameters in broiler after 15, 30, and 45 days of treatments

Values using various crafts in the pillar are significantly diverse at P<0.05.

The data in table 4 denote a significant rise in the level of SOD for the two BHA groups during 30 and 45 days compared with the control. As for the results of CAT, the results indicated a significant increase in the BHA group with a dose of 5 mg within 45 days compared with the control and BHA groups at a rate of 2.5 mg, the BHA group with a dose of 5 mg during the 30 days of the study and the BHA group with a dose of 2.5 mg during the periods of 30 and 45 days indicate a significant elevate in the level of CAT compared with the control. Treatment with the two groups of BHA caused a significant increase in the rate of T3 during 30 and 45 days of the study compared with the control, and the BHA group with a dose of 5 mg illustrate a significant rise in the level of T3 compared to the BHA group at a rate of 2.5 mg during 30 days of the experiment. Treatment with both BHA groups led to a significant rise in T4 levels during the 30 and 45 days of the study compared with the control.

Table 4: Influence of BHA on some antioxidant status and thyroid hormone in broiler after 15, 30, and 45 days of treatments

Values using various crafts in the pillar are significantly diverse at P<0.05.

Discussion

The results obtained noted that BHA led to an improvement in broilers' physiological and blood characteristics. Although it is considered one of the widely used industrial phenolic antioxidants as feed additives in poultry diets (11), scientific research on its essential effects in many physiological traits was not widely recorded. It is noted that the addition of BHA to the broiler diet with different concentrations led to an elevated rate of lymphocytes accompanied by a reduction in the rate of heterophils, eosinophils, and basophils. This effect in the number of blood cells can be attributed to its effect on improving the immune status of the bird, as Lanzhou et al. (12) indicated that the addition of BHA to the diets of broilers at the age of 1-21 days at a concentration of 5 and 10 mg/kg of feed led to an improvement in the bird’s immune status. Treatment with BHA caused an excess in the number of red and white cells, and this is consistent with what was mentioned by Adeyemi et al. (13), where he indicated that adding BHA at a concentration of 0.12 g/kg in broiler diets led to an increase in the numbers of these cells. The effect of BHA on some hematological characteristics can also be attributed to thyroid hormones. These hormones play an essential role in the progress of lymphoid organs and the arranging of immune assignment in birds and raise the number of lymphocytes, phagocytes, IgM, and IgG (14), as indicated by Keshavarz et al. (15) to giving thyroxine at a concentration of 0.3 mg/bird daily to laying hens at the age of 47 weeks for 14 weeks led to an increase in the number of white and red cells, and this was identical to the results of this study. The increase in hemoglobin and packed cell volume is due to a direct relationship with an upraise in the number of RBC. The results also showed a significant reduction in MCV level due to an inverse relationship with an increase in red cells.

In contrast, the significant high in the level of MCHC due to using different concentrations of BHA could be attributed to a direct relationship with the observed increase in the level of hemoglobin (16). The data illustrate a significant rise in antioxidants represented by an increase in SOD and CAT in the serum of broilers. This increase can be attributed to its effect on improving the internal environment of the bird, as Kh et al. (17) indicated that the gastrointestinal tract is the leading site for taking nutrients from the external environment. It plays an essential role in reducing the oxidative stress produced by food oxidation. This is similar to what Barsha et al., Xiaoqing et al. (5,18) mentioned. They stated that BHA has a vital role in inhibiting the oxidation process of fats by increasing the effectiveness of antioxidants and has a significant role in protecting the liver from oxidative stress and by increasing the level of CAT, SOD, ALT AST, and glutathione and decreasing the level of malondialdehyde when added at a dose of 0.5 mg/kg to the diet of broilers. The results also noted a significant upraise in the level of thyroid hormones, and no study was recorded on the effect of BHA on the level of these hormones. However, it is believed that its role in this increase can be attributed to improving the health of the bird through increasing the level of SOD and CAT, Which is one of the first lines of defense in the body (19), where superoxide dismutase works to break down free radicals and convert them to hydrogen peroxide. In contrast, catalase works to convert hydrogen peroxide into water and oxygen (20). Therefore, it is noted through these results that BHA is one of the essential antioxidants that are used as feed additives in poultry diets. It has physiological effects on some hematological and hormonal parameters.

Conclusion

The results obtained conclude that industrial antioxidants like BHA have significant effects in improving broilers' physiological and hormonal characteristics when used as feed additives in poultry diets.

Acknowledgments

The researcher extends thanks and appreciation to the University of Mosul and the College of Veterinary Medicine for contributing to the completion of this research.

Conflict of interest

The researcher indicates that there are no conflicts of benefit related to the publication of this study.