Research Article

Effect of Inorganic Selenium on Blood Biochemistry under Dexamethasone Induced Stress in Broiler Chicken

Irfan Ullah1, Asad Ullah2*, Tahira Tayyeb1, Rafiq Ullah1, Muhammad Hanif1, Faiza Khan3, Imad Khan2, Raheela Taj4, Fatima Syed4, Shumaila Gul5, Muhammad Sadeeq6, Muneeb Islam7, Arsalan Khan8 and Khudija Ghani9

1Department of Zoology, Abdul Wali Khan University Mardan, (KP), Pakistan; 2College of Veterinary Science and Animal Husbandry (CVS and AH), Abdul Wali Khan University Mardan (KP), Pakistan; 3Department of Radiology, Hayatabad Medical Complex Peshawar (KP) Pakistan; 4Institute of Chemical Sciences (ICS), University of Peshawar (KP), Pakistan; 5Department of Chemical and Life Sciences, Qurtuba University of Science and Information, Technology Peshawar (KP), Pakistan; 6University of Veterinary and Animal Sciences, Swat (KP), Pakistan; 7Department of Microbiology, Abdul Wali Khan University Mardan (KP), Pakistan; 8Veterinary Research and Disease Investigation Centre (VR and DIC), Dera Ismail Khan (KP), Pakistan; 9Sarhad Institute of Allied Health Sciences, Sarhad University of Science and Information Technology, Peshawar (KP), Pakistan.

Received | October 19, 2023; Accepted | December 20, 2023; Published | February 13, 2024

*Correspondence | Asad Ullah, College of Veterinary Sciences and Animal Husbandry, Abdul Wali Khan University, Mardan 23200, Khyber Pakhtunkhwa, Pakistan; Email: [email protected]

Citation | Ullah, I., A. Ullah, T. Tayyeb, R. Ullah, M. Hanif, F. Khan, I. Khan, R. Taj, F. Syed, S. Gul, M. Sadeeq, M. Islam, A. Khan and K. Ghani. 2024. Effect of inorganic selenium on blood biochemistry under dexamethasone induced stress in broiler chicken. Sarhad Journal of Agriculture, 40(1): 142-148.

DOI | https://dx.doi.org/10.17582/journal.sja/2024/40.1.142.148

Keywords | Broiler, Selenium, Dexamethasone, Glucocorticoid, Biochemistry, Enzyme

Copyright: 2024 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

The poultry industry has many challenges to suppressed bird’s immunity, growth and production performance. The emergence and re-emergence of diseases user confidence type of product safety are the major challenges to the current situation for the future of national and international industry (Hafez and Attia, 2020). There is huge pressure on poultry, market to establish equilibrium between demand and supply. Several types of stresses like transport, fasting, temperature, overcrowding and antibiotics etc., are the main causes behind this gap. These stresses affect the growth as well health of the broilers and sometime cause mortality (Hussain et al., 2015). In every stress condition the level of glucocorticoid suddenly arise. Both natural and synthetic derivatives steroid hormones are present in Glucocorticoids (GCs) which can cause several damages to birds like effecting kidney function, liver enzymatic production increase in blood creatinine, urea and uric acid and suppress the immune system (Duff et al., 2019). DEX is a synthetic form of the glucocorticoids. Therefore, the antibiotics growth promoters are banned internationally to protect the human health and environment. The use of alternative of antibiotics growth promoter is needed to enhance the health status of the birds. It has been reported that the commercially available alternative phytobiotic, probiotic and trace minerals have impact on the health, production and performance of poultry (Ren et al., 2019). Trace minerals (TMs) play important role in production of broiler chicks, it not only play role in health maintains but also in quality of meat production control of disease and reduce stress. TMs are also function on various biosynthetic, digestive and physiological process as a co-factors of several metallo-enzymes, including the MN-SOD, Zn/Cu-SOD and selenium containing enzymes (Wang et al., 2019). Currently inorganic selenium (Se) trace element attract the more attention of poultry farmer because of strong adsorbing ability, high bioavailability, low toxicity and high catalytic efficiency as compare with organic selenium on broiler poultry (Wang et al., 2019). Selenium is the important trace mineral element that needed for the body normal function and therefore on the broiler chickens and have important role in their health maintenance. The sign and symptom of Se deficiency on poultry has been a vital role in protection of antioxidant through the enzyme glutathione peroxidase (GPx). It has been recommended that the selenium supplemented feed of roosters can help as a nutritional food have low cholesterol and high selenium substances for both cardiovascular disorder patient and healthy people. Selenium is an essential trace mineral that plays vital role in productivity, growth, immune function and anti-stress. The different forms of selenium supplementation such as organic and inorganic selenium the concentration of selenium on broiler diet is different. In nature selenium is present in two chemical form inorganic and organic. Inorganic selenium can be found in several minerals in the form of, selenite, selenide and also in the metallic form. In other side oilseed, grain and forages meals the selenium is bond to the different amino acid such as cysteine and methionine. Therefore, the animal in nature receives the selenium primarily as seleno-methionine The majority of the selenium absorbed by the plant is in the form of selenite or selenate, or it is produced by the plant using seleno amino acids, which account for at least 50% of the selenium in cereal grains. Plants are Se-methyl-Selenocysteine, Selenocysteine, and Se-methyl-Seleno-methionine (Surai, 2002). However, the use of selenium on broiler industries is comparatively less therefore the current study was designed to find the impact of inorganic se supplementation, on the serum biochemistry in broiler chickens under dexamethasone induced stress. To examine the effect of inorganic Se supplementation on liver enzyme (ALT, AST, ALP), renal function (blood urea, creatinine and uric acid), lipid profile i.e; Total Cholesterol, High density lipoprotein (HDL) and Low density lipoprotein (LDL) and blood glucose level on broiler chickens under dexamethasone induced stress.

Materials and Methods

Experimental design, birds and rearing conditions

The trial was conduct in experimental poultry shed at College of Veterinary Sciences and Animal Husbandry (CVS and AH), Abdul Wali khan University, Mardan. The collected samples were processed in laboratory of physiology, CVS and AH. Two-hundred-day-old birds were allocated to five groups and having eight replicates in each group and five (5) birds per replica. The total duration of the experimental trial was five weeks (35 days), and the birds were kept in an environmentally controlled experimental shed. Temperature of 35℃ was maintained on the day 1st and was decreased gradually (3℃/week) to 26℃ by the end of the 3rd week (21 day). From day 22nd to the end of the trial, the temperature was kept at 26℃. The relative humidity (RH) was kept at 65% during the whole experiment. Group-A was considered as a negative control group provided with basal diet (BD) only, Group-B was fed with BD and dexamethasone (DEX-15mg/kg), Group-C was supplemented with 0.2mg Se+DEX-15mg/kg in BD. Similarly, Group-D was supplemented with 0.3mg Se+DEX-15mg/kg) in BD and Group-E was fed with BD+0.4mgSe+15mg DEX/kg (Table 1). The freshwater was offered ad libitum during the whole research trial.

Sampling

Two chicks from each replicate were randomly selected and slaughtered. The blood was collected from jugular veins in vacutainer and centrifuged for 15 minutes at 1000 rpm for serum analysis. The parameters under observation were liver function test (ALT, AST, and ALP) and kidney function (blood urea, uric acid, creatinine, and blood glucose and lipid profile.

Urea, creatinine, uric acid, and glucose were determined from blood serum through available commercial kit. A total of 20µ liter of serum was taken in a calcium glass tube with the help of micropipette for determination of Urea. Then 1000µ liter of reagent 01 and 250µ liter of reagent 02 was mixed with it using DiaSys–Germany REF 12767726876. After mixing, the mixture was incubated for 50 seconds and the results were displayed on the screen and the values were noted. Creatinine assay kit was used for determination of creatinine and 50µ liter of the serum was obtained in calcium glass tube. Then, 1000µ liter of reagent 01 and 250µ liter of reagent 02 was mixed with it using DiaSys–Germany REF 12767726876. The tube containing mixture was incubated for 60 seconds and the obtained results were noted. Glucose was determined by taking 10µ liter serum and 1000µ liter of reagent 01 with 250µ liter of reagent 02 in calcium glass tube and was mixed using DiaSys–Germany REF 12767726876. The tube containing mixture was incubated for 5 minutes and the obtained results were noted

Statistical analysis

Statistical analysis was done through SPSS software (IBM SPSS Statistics 22), respectively and subjected to one-way ANOVA and displayed as mean standard error mean (SEM) (Version 20.0). The one-way analysis of variance (ANOVA) was employed to examine the means of different groups in the study. The Duncan post hoc test was employed to evaluate the disparities between groups, with the critical level set at a significance level of (P ≤ 0.05).

Results and Discussion

According to the result of the current study, the values of ALT were significant (P≤0.05) in negative control group (A) as compare to the group C, D and E. While the value of AST is also higher (P≤0.05) in group A (negative control) as compare to the value of C, D and E. The values of ALP were competitively high (P≤0.05) in group A (negative control) (P≤0.05) as compared to group C, D and E (Table 2).

 

Table 1: Feed composition of different groups during research trial.

Groups	A	B	C	D	E
Feed composition	BD	BD+DEX-15mg/kg	0.2mg Se+DEX-15mg/kg	0.3mg Se+DEX-15mg/kg	0.4mg Se+DEX-15mg/kg

BD: Basal diet; Se: Selenium; DEX: Dexamethasone.

 

Table 2: Effect of selenium (Se) on liver enzymatic function (ALT, AST, ALP) of broiler chickens under dexamethasone induced stress.

Groups	A	B	C	D	E	SEM	P-Value
ALT(µ/l)	7.92bc	13.90a	10.62b	5.75d	6.62cd	0.418	0.02
AST (µ/l)	152.80b	300.62a	151.30b	150.60b	153.10b	5.59	0.03
ALP (µ/l)	1440.50a	1802.2b	1510.25a	1480.70a	1501.0a	32.15	0.02
	* − data statistically significant at P<0.05

ALT: Alanine transaminase; AST: Aspartate aminotransferase; ALP: Alkaline phosphatase.

 

Table 3: Effect of selenium (Se) on renal function (urea, uric acid, creatinine, glucose level) of broiler chicken under dexamethasone induced stress.

Groups	A	B	C	D	E	SEM	p-value
UC(mg/dl)	4.33b	5.562a	4.82b	4.65b	4.31 b	0.0332	0.02
CrT (mg/dl)	0.20 b	0.287a	0.175b	0.20 b	0.162b	0.0154	0.01
U(mg/dl)	5.25b	7.50a	5.25 b	4.62b	4.37 b	0.4304	0.04
G (mg/dl)	177.46bc	190.20a	178.0b	172.4bc	152.36c	4.614	0.03

UC: Uric Acid; CRT: Creatinine; U: Urea; G: Glucose.

 

Table 4: Effect of selenium (Se) on lipid profile of broiler chicken under dexamethasone induced stress.

Parameter	A	B	C	D	E	SEM	P-Value
Cholesterol (Mg/dl	127.00ab	138.20a	128.70ab	118.30b	113.30 b	2.44	0.026
Triglycerides (mg/dl)	59.30	62.80	60.25	72.66	70.42	2.84	0.344
HDL (mg/dl)	68.80	66.50	70.26	74.40	74.30	3.284	0.200
LDL (mg/dl)	86.38b	99.58a	89.75b	83.36b	88.42b	2.478	0.008

HDL: high density lipoprotein; LDL: low density lipoprotein.

 

The result of current study concludes that the level of uric acid in blood was significantly higher (P≤0.05) in group A (negative control) (P≤0.05) as compared to the group B (positive control), C, D, and E group. While the blood creatinine level was also higher (P≤0.05) in group A (negative control group) than group B (positive control) and selenium supplementation group C, D and E. The blood urea was also comparatively high in group A (negative control) (P≤0.05) than positive control group B and selenium supplemented group C, D and E. The glucose concentration in blood was also higher (P≤0.05) in group A (negative control) than the group B (positive control) and group C, D and E (Table 3).

 

The lipid profile result showed that the blood cholesterol level in group A (negative control) significantly (P≤0.00) higher than the group B (positive control) and SE supplemented group C, D and E. The values of triglycerides were remained insignificant. The level of HDL was also recorded insignificant. The level of LDL in blood of broiler is higher (P≤0.05) in DEX treated group than the negative control group A and Se supplemented groups C, D and E (Table 4).

This study was designed to analyze the effect of selenium supplementation source of trace mineral and their relation on the antioxidant enzymatic activity, growth performance and serum concentration in the broiler chicken. Selenium is considered as a vital trace mineral play important role in growth performance antioxidant status and liver enzymatic activity. The current study showed that the ALT level is increase in the group B (positive control or DEX treated group) while the ALT level in group C, D and E is decrease due to selenium supplementation. Similarly, the level of AST and ALP also increase in positive control group B (DEX treated group) while their level decrease in Se supplemented group C, D and E. The previous study found that the Se supplementation improve the antioxidant activity, growth performance, liver and kidney function of broiler chickens. Another study of Boostani et al. (2014) supported our current study, to show the effect of selenium on serum biochemistry. They concluded that the Se supplements increase the liver enzymatic function, renal function and growth performance of broiler chickens.

In present study we investigate that the addition of Se on feed of broiler chicken can positively effect on the kidney function, blood urea level, level of uric acid, creatinine and glucose in blood. Selenium is a trace mineral plays important role in growth performance liver and kidney function and blood glucose concentration. The current study showed that the uric acid level was increased in positive control group B (DEX treated group) while the level uric acid is decreased in Se supplemented group C, D and E. The same results were also reported by Zhang et al. (2015) and Liang et al. (2015) who suggested that the level of uric acid was reduced with the selenium supplementation in broilers. According to current study, the level of blood urea and creatinine level in blood were increased in DEX treated group while level of the same was remained normal in negative control group A. However, the level of urea and creatinine was become down in Se supplemented group C, D and E. The same study was conduct by Sun et al. (2015) who reported that the deficiency of selenium can increase the level creatinine and urea in blood while decrease the renal antioxidant capacity to control the level of blood urea and creatinine and sometime the selenium deficiency and use of high level of glucocorticoid increase the risk of damages in renal tissues due to which the level of urea and creatinine increase in blood.

In the current research findings, the blood cholesterol level in group B (positive control) was significantly higher in comparison to group A (negative control). The level of LDL in blood of broiler was higher (P≤0.05) in DEX treated group (positive control) than the negative control (group A) and Se supplemented groups. Dexamethasone induced oxidative stress and increased the glucocorticoid, cortisol and cholesterol level in blood circulation. Selenium supplements in the diet of broilers enhance the antioxidant activity and reduce the level of cholesterol when compared with dexamethasone treated group (Nwakpu et al., 2016). Selenium are important trace mineral and improved the activity of antioxidant enzyme such as catalase, Plasma glutathione peroxidase (GHS-PX), Superoxide dismutase (SOD), HDL-Cholesterol and decrease serum cholesterol, Low density lipoprotein (LDL)-Cholesterol and Malondialdehyde (MDA) level in the broiler (Cai et al., 2012).

Conclusions and Recommendations

In the current research investigation, DEX increased the serum creatinine level in chicks. The same has been reported in case of urea and uric acid and could be attributed to an initial state of stress induced by DEX affecting the kidneys. Similarly, the level of cholesterol was high in DEX treated chicks and lowered in negative control and chicks treated with Se supplementation. Similar to our findings that reduced serum creatinine levels was noted in birds when supplemented with Selenium is also reported previously. Selenium improves the level blood urea and uric acid. Similarly, Se due to its hypolipidemic properties increased the level of triglyceride while DEX induced birds showed decreased level of HDL. The serum cholesterol levels were also high in DEX treated birds compared to negative control group and selenium treated birds. Use of DEX also increased the level of glucose in blood of broiler and Se reduced the level of glucose in birds. Similarly, the level of ALT, AST and ALP is also increased by the use of DEX in broiler. While the supplementation of selenium decreased the level of ALT, AST and reduced the ALP significantly. It is recommended that further research work needs to be carried out on the effect of selenium (Se) on the broiler growth and health. Further, different awareness seminars and workshops are necessary for the poultry farmer about the addition of selenium in broiler feed. The researcher must use selenium on deferent animals and birds with different concentrations and show their effect and outcomes to the farmers, academia and industry.

Acknowledgments

The authors are thankful to the supporting staff of both the Department of Zoology and College of Veterinary Science and Animal Husbandry (CVS and AH), Abdul Wali Khan University Mardan, Khyber Pakhtunkhwa for their assistance during research trial.

Novelty Statement

The research and experimental work on the subject title is original and new in the field of poultry feed supplementation in Khyber Pakhtunkhwa, Pakistan.

Author’s Contribution

Irfan Ullah: Investigation, writing-original draft preparation.

Asad Ullah: Conceptualization, supervision.

Tahira Tayyeb: Methodology.

Rafiq Ullah: Project administration.

Muhammad Hanif and Faiza Khan: Validation.

Imad Khan: Resources.

Raheela Taj: Data curation.

Fatima Syed: Visualization.

Shumaila Gul: Formal analysis.

Muhammad Sadeeq: Software.

Muneeb Islam and Arsalan Khan: Writing-review and editing.

Khudija Ghani: Helped in collection of relevant literature.

Ethical approval

The study was approved by the ethical review committee, Department of Zoology, Abdul Wali Khan University Mardan.

Conflict of interest

The authors have declared no conflict of interest.

References

Arnaut, P.R., G. da Silva Viana, L. da Fonseca, W.J. Alves, J.C.L. Muniz, J.E. Pettigrew, H.S. Rostagno and M.I. Hannas. 2021. Selenium source and level on performance, selenium retention and biochemical responses of young broiler chicks. BMC Vet. Resear., 17(1): 1-13. https://doi.org/10.1186/s12917-021-02855-4

Boostani, A., A.A. Sadeghi, S.N. Mousavi, M. Chamani and N. Kashan. 2015. The effects of organic, inorganic, and Nano-selenium on blood attributes in broiler chickens exposed to oxidative stress. Acta Sci. Vet., 43: 1-6. https://www.ufrgs.br/actavet

Cai, S.J., C.X. Wu, L.M. Gong, T. Song, H. Wu and L.Y. Zhang. 2012. Effects of nano-selenium on performance, meat quality, immune function, oxidation resistance, and tissue selenium content in broilers. Poult. Sci., 91(2): 2532–2539. https://doi.org/10.3382/ps.2012-02160

Chen, G., J. Wu and C. Li. 2014. Effect of different selenium sources on production performance and biochemical parameters of broilers. J. Anim. Physiol. Anim. Nutr., 98(4): 747-754. https://doi.org/10.1111/jpn.12136

Duff, A.F., M.F. Baxter, B.D. Graham, B.M. Hargis and L.R. Bielke. 2019. Mode of action of dietary dexamethasone may not be dependent upon microbial mechanisms in broilers. Microorgan, 7(9): 346. https://doi.org/10.3390/microorganisms7090346

Eid, Y., T. Ebeid and H. Younis. 2006. Vitamin E supplementation reduces dexamethasone-induced oxidative stress in chicken semen. Br. Poult. Sci., 47(3): 350-356. https://doi.org/10.1080/00071660600753912

Eid, Y., T. Ebeid, M. Moawad and M. El-Habbak. 2008. Reduction of dexamethasone-induced oxidative stress and lipid peroxidation in laying hens by dietary vitamin E supplementation. Emi. J. F. Agric., 2: 28-40. https://doi.org/10.9755/ejfa.v20i2.5188

Fan, C., B. Yu and D. Chen. 2009. Effects of different sources and levels of selenium on performance, thyroid function and antioxidant status in stressed broiler chickens. Int. J. Poult. Sci., 8(6): 583-587. https://doi.org/10.3923/ijps.2009.583.587

Hafez, H.M. and Y.A. Attia. 2020. Challenges to the poultry industry: Current perspectives and strategic future after the COVID-19 outbreak. Front. Vet. Sci., 7: 516. https://doi.org/10.3389/fvets.2020.00516

Hussain, J., I. Rabbani, S. Aslam and H.A. Ahmad. 2015. An overview of poultry industry in Pakistan. World Poult. Sci. J., 71(4): 689-700. https://doi.org/10.1017/S0043933915002366

Khoobani, M., S.H. Hasheminezhad, F. Javandel, M. Nosrati, A. Seidavi, I.T. Kadim, V. Laudadio and V. Tufarelli. 2019. Effects of dietary chicory (Chicorium intybus L.) and probiotic blend as natural feed additives on performance traits, blood biochemistry, and gut micro biota of broiler chickens. Antibiot. 9(1): 5. https://doi.org/10.3390/antibiotics9010005

Liang, N., F. Wang, X. Peng, J. Fang, H. Cui, Z. Chen, W. Lai, Y. Zhou and Y. Geng. 2015. Effect of sodium selenite on pathological changes and renal functions in broilers fed a diet containing aflatoxin B1. Int. J. Environ. Res. Publ. Health, 12(9): 11196-11208. https://doi.org/10.3390/ijerph120911196

Nwakpu, P.E. Osita, C.O. Nweke. 2016. Effect of dietary selenium suppression on dexamethasone induced oxidative stress on the growth performance and haematology of broiler chickens. Int. J. Agric. Environ. Res., 2(4): 857-868.

Papp, L.V., J. Lu, A. Holmgren and K.K. Khanna. 2007. From selenium to selenoproteins: synthesis, identity, and their role in human health. Antioxid. Redox Signal, 9(7): 775-806. https://doi.org/10.1089/ars.2007.1528

Petrovič, V., L. Nollet and G. Kováč. 2010. Effect of dietary supplementation of trace elements on the growth performance and their distribution in the breast and thigh muscles depending on the age of broiler chickens. Acta Vet. Brno., 79(2): 203-209. https://doi.org/10.2754/avb201079020203

Ren, H., W. Vahjen, T. Dadi, E.M. Saliu, F.G. Boroojeni and J. Zentek. 2019. Synergistic effects of probiotics and photobiotic on the intestinal micro biota in young broiler chicken. Microorga. 7(12): 684. https://doi.org/10.3390/microorganisms7120684

Sharifi, S.D., A.A. Khadem, E. Tavazoee, A. Barin and H. Moslehi. 2010. Effects of Flavomycin and protexin on growth performance and intestinal microflora population of broiler fed diets with different levels of soy oil. Krmiva: Časopis o hranidbi životinja, Proizv. Tehnol. Kr. 52(3): 153-156.

Sultana, N., M. Afrose and K. Rafiq. 2020. Effects of steroid growth promoter on morphological and biochemical adaptations in liver of broiler. Vet. World, 13(11): 2330. https://doi.org/10.14202/vetworld.2020.2330-2337

Sultana, N., R. Islam, A. Akter, U. Ayman, S. Bhakta, S.A. Rony, A. Nahar and R. Alam. 2021. Biochemical and morphological attributes of broiler kidney in response to dietary glucocorticoid, dexamethasone. Saudi J. Biol. Sci., 28(12): 6721-6729. https://doi.org/10.1016/j.sjbs.2021.07.047

Sun, D., C. Li, J. Gao, S. Li and H. Wang. 2015. Effects of selenium deficiency on principal indexes of chicken kidney function. Biol. T. Element. Res., 164(1): 58-63. https://doi.org/10.1007/s12011-014-0196-0

Surai, P.F., 2002. Selenium in poultry nutrition 1. Antioxidant properties, deficiency and toxicity. World Poult. Sci. J., 58(3): 333-347. https://doi.org/10.1079/WPS20020026

Vicuña, E.A., V.A. Kuttappan, R. Galarza-Seeber, J.D. Latorre, O.B. Faulkner, B.M. Hargis, G. Tellez and L.R. Bielke. 2015. Effect of dexamethasone in feed on intestinal permeability, differential white blood cell counts, and immune organs in broiler chicks. Poult. Sci., 94(9): 2075-2080. https://doi.org/10.3382/ps/pev211

Wang, G., L.J. Liu, W.J. Tao, Z.P. Xiao, X. Pei, B.J. Liu, M.Q. Wang, G. Lin and T.Y. Ao. 2019. Effects of replacing inorganic trace minerals with organic trace minerals on the production performance, blood profiles, and antioxidant status of broiler breeders. Poult. Sci., 98(7): 2888-2895. https://doi.org/10.3382/ps/pez035

Zhang, J.L., B. Xu, X.D. Huang, Y.H. Gao, Y. Chen and A.S. Shan. 2016. Selenium deficiency affects the mRNA expression of inflammatory factors and selenoprotein genes in the kidneys of broiler chicks. Biol. Trace Element. Res. 171(1): 201-207. https://doi.org/10.1007/s12011-015-0512-3