Thyme, Celery and Salinomycin Implication on Antioxidant Capacity and Neurotransmitters Related to Milk Production in Pregnant Barki Ewes

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Thyme, Celery and Salinomycin Implication on Antioxidant Capacity and Neurotransmitters Related to Milk Production in Pregnant Barki Ewes Advances in Applied Physiology 2021; 6(1): 23-29 http://www.sciencepublishinggroup.com/j/aap doi: 10.11648/j.aap.20210601.14 ISSN: 2471-9692 (Print); ISSN: 2471-9714 (Online) Thyme, Celery and Salinomycin Implication on Antioxidant Capacity and Neurotransmitters Related to Milk Production in Pregnant Barki Ewes Sherif Yousif Eid 1, *, Omar Abdel Hamid Ahmed-Farid 2, Hussein Mostafa El-Zaher 1, 1 Mahmoud Mohammed Shabaan 1Biological Applications Department, Egyptian Atomic Energy Authority (EAEA), Cairo, Egypt 2Physiology Department, National Organization for Drug Control and Research (NODCAR), Cairo, Egypt Email address: *Corresponding author To cite this article: Sherif Yousif Eid, Omar Abdel Hamid Ahmed-Farid, Hussein Mostafa El-Zaher, Mahmoud Mohammed Shabaan. Thyme, Celery and Salinomycin Implication on Antioxidant Capacity and Neurotransmitters Related to Milk Production in Pregnant Barki Ewes. Advances in Applied Physiology. Vol. 6, No. 1, 2021, pp. 23-29. doi: 10.11648/j.aap.20210601.14 Received : May 7, 2021; Accepted : May 25, 2021; Published : May 31, 2021 Abstract: The experiment goal was the investigation of thyme (T), celery (C) and salinomycin effects on immune response, neurotransmitters related to milk production in Barki ewes. Total 72 mature ewes (2-3 years & 40±1.5 Kg BW) randomly pined equally into five groups. Group-1 was control; groups 2 & 3 received 20g/head/day T and C, respectively. Group-4 received 10g T+ 10g C/head/day, group-5 treated with salinomycin 1g/head/day. Samples collected during 2nd , 3rd trimester of pregnancy and on delivery day (DD); milk yield assessed on 15, 30 and 45-day postpartum. T and/or C and salinomycin increased (P<0.05) superoxide dismutase (SOD), reduced glutathione (GSH) and glutathione disulfide (GSSG) during mid-, late-pregnancy and DD compared to control, celery and thyme increase malondialdehyde (MDA) (p<0.05) during mid-and late-pregnancy, respectively compared to other groups. Nitric oxide (NO) levels increased in thyme X celery (TxC) group during mid-pregnancy and DD with insignificantly compared with other groups. During mid-pregnancy TxC treatment increased (p<0.05) serotonin (5-HT) levels compared with other groups, the same was dopamine (DA), norepinephrine (NE) and tryptophan (Trp) levels (P>0.05). During late pregnancy 5-HT, DA, NE & Trp increased (P<0.05) in the thyme and/or celery group. While on DD salinomycin increased neurotransmitters (P<0.05) with an insignificant increase in other groups. Milk yield increased (P<0.05) during 15, 30 and 45 days postpartum in T and/or C groups than control and salinomycin. In conclusion, the applied treatments had a significant effect on reproductive performance, immune response in ewes throughout pregnancy and DD periods, and milk production during the postpartum period. Keywords: Thyme, Celery, Salinomycin, Immune System, Neurotransmitters, Milk Yield, Ewes milk components [1, 2]. Medicinal plants and ionophores had 1. Introduction a wide consideration as feed additives that improve farm Pregnancy sustainability and increase milk yield after animals’ antioxidant status and ameliorate physiological parturition are vital goals for animal breeders that guarantee stress generated from changes in the reproductive (i.e., the normal and healthy growth of newborn offspring. Milk pregnancy and/or parturition) or productive (lactation) stage production is a unique physiologic completion of the because of their chemical composition and functional groups; reproductive cycle that is regulated by an array of interacting such as phenols, thymol, carvacrol, terpenes [3] for thyme; endo- and neurocrine process. prolactin (PRL) is the most and mono, bi, and sesquiterpenes hydrocarbons [4] for Celery, hormone required for lactogenesis, which provides a that gives them the antioxidant, and antimicrobial potentiality comprehensive signal that fosters synthesis and secretion of [5-7]. Celery supplementation reduced glutathione, catalase, 24 Sherif Yousif Eid et al .: Thyme, Celery and Salinomycin Implication on Antioxidant Capacity and Neurotransmitters Related to Milk Production in Pregnant Barki Ewes lipid peroxidation [8], and showed a suppressive effect Cairo, Egypt. against malonaldehyde generation [9]. Salinomycin is a fermentation product of various Streptomyces spp used on 2.1. Herbal Plants large scale as a growth stimulus as well as animal health and Thyme (dried leaves) and seeds of celery were brought welfare enhancer via the shedding of pathogens [10]. Rare from HARAZ herbal store, Cairo, Egypt. Then ground investigations have examined the effect of T, C and separately and used in the study. salinomycin on reproductive performance and sexual- immune response throughout pregnancy and parturition in 2.2. Experiental Animal Management and Treatments sheep, rather than milk production during the postpartum period. The objective here was to examine T, C, and Total of 72 mature ewes (2-3 years & 40±1.5 Kg mean Salinomycin effects on neurotransmitters related to milk body weight) were randomly pined equally into 5 groups as production, antioxidant capacity and sexual hormone profile in Table 1. The concentrate feed mixture (CFM) used in the in ewes throughout pregnancy and delivery day, and milk feeding trial of ewes consisted of (as a percentage): 24.0 corn, production during the postpartum period. 22.5 sugar beet pulp, 20.0 wheat bran, 30.0 undecorticated cotton seed cake, 1.0 sodium chloride, 0.3 mineral mix, 2.0 dicalcium phosphate, 0.1 AD 3E, and 0.1 sodium bicarbonate. 2. Materials and Methods Ewes were fed twice daily at 07:00 and 21:00 hrs. Fresh The current study was conducted in sheep experimental drinking water was available at all times. The animals farm, Nuclear Research Center, Egyptian Atomic Energy received treatments 6weeks before breeding and continued Authority. The study was approved by the EAEA Committee, until the end of the experiment. Table 1. The experimental design used in feeding the study. Control Treated groups Feedstuffs program G1 G2 G3 G4 G5 CFM, % 60 60 60 60 60 Berseem % 40 40 40 40 40 Thyme (T) --- 20g/h/d --- --- --- Celery (C) --- --- 20g/h/d --- --- Thyme × Celery (TxC) --- --- --- 10×10g/h/d --- Salinomycin (S) --- --- --- --- 1g/h/d G1=control, G2=Thyme, G3=Celery, G4=TxC, G5=Salinomycin and CFM=Concentrate feed mixture. 10 _m, mobile phase 45:55 of 0.1 M NaCl-methanol); 2.3. Blood Sampling, Milk Yield, and Hormones Analysis wavelength was adjusted to 230 nm, according to HPLC Evacuated non-heparinized glass tubes were used to procedure [12]. collect blood samples from the jugular vein, then left at room 2.5. Determination of the Neurotransmitter Concentrations temperature from 30 to 60 min for clotting. Later the tubes by HPLC Method centrifuged at 3000 rpm for 15 min to harvest serum. The latest was stocked up on -70°C until analysis. Samples were The HPLC system consisted of a quaternary pump; a taken during the 2nd and 3rd trimester of pregnancy as well as column oven, Rheodine injector and 20µl loop, UV variable on delivery day (DD). Milk yield was recorded for wavelength detector. The report and chromatogram were individuals once every two weeks started from the second taken from the data acquisition program purchased from week until 8 weeks of lactation. Twenty-four hours before Chemstation. The sample was immediately extracted from hand milking, the lambs were kept away from their dams. the trace elements and lipids by the use of solid-phase Ewes were completely hand milked. Blood E2-17β (pg/ml) & extraction CHROMABOND column NH2 phase cat. No. P4 (ng/ml) levels were determined using ELISA kits (Cat. No. 730031. The sample was then injected directly into an MBS734529; MBS269207, respectively). AQUA column 150 mm 5µ C18, purchased from Phenomenex, USA under the following conditions: mobile 2.4. Antioxidant’s Capacity Estimations phase 20mM potassium phosphate, pH 2.5, flow rate The activities of GSH, GSSG were determined 1.5ml/min, UV 190 nm. Noradrenalin, dopamine, and according to the procedure of chromatographic method serotonin were separated after 12 minutes . The resulting with standard Sigma-Aldrich Cat. No. 70-18-8, 27025-41- chromatogram identified each monoamine position and 8, respectively [11]. MDA and SOD serum contents were concentration from the sample as compared to that of the assessed following kits' instruction of Bio-diagnostic® standard, and finally, the determination of the content of each firm (Cairo, Egypt) (Cat. No: SD 2521 & MD 2529, monoamine as µg per mL [13]. respectively). Nitric oxide was calculated as the sum of 2.6. Statistical Analysis nitrite and nitrate by using anion exchange column (Hamilton PRP-X100) using the parameters (150*4.1mm, Data were expressed as mean±SE. Statistical analysis of Advances in Applied Physiology 2021; 6(1): 23-29 25 the obtained data was performed using the general linear 0.542 ng/mL; P<0.01 at parturition (Figure 2). T, C and TxC model (GLM) using SAS software [14]. Significant groups were higher (P<0.01) in E2 levels than control at late- differences among means were evaluated using Duncan’s pregnancy and DD, the highest E2 mean value was 12.38 Multiple Range Test. Pg/mL for T treatment on DD. During late pregnancy, only TxC group recorded an increase (P<0.01) on serum P4 than 3. Results control, while at mid-pregnancy P4 levels increased (P<0.01) than control due to treatments (except Salinomycin) with the C, T and TxC treatments increased (P<0.01) SOD highest mean value 4.11 ng/mL for Celery. No significant activity more than control during the second and third changes in P4 levels than control on DD (Table 4). trimester of pregnancy and DD with the highest values of As shown in Table 5, ewes’ supplementation with thyme 223.6, 215.0 & 210 U/mL, respectively through the 3rd and/or celery showed a significant increase (P<0.001) in total trimester.
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