Profiles of Blood Amino, Fatty and Volatile Acids in Awassi Ewes Across Different Reproductive Stages and Their Association with Litter Size and Offspring Sex
Keywords:
amino acids, fatty acids, volatile fatty acids, twins, pregnancy, sheepAbstract
This study aimed to investigate the dynamic changes in the blood profiles of amino acids, fatty acids, and volatile acids in Iraqi Awassi ewes across different reproductive stages, and to evaluate their associations with litter size and offspring sex. A total of thirty mature ewes were selected. Blood samples were collected via jugular venipuncture at five key reproductive stages: day 0 (pre-mating), 14 days post-mating, and at 45, 75, and 135 days of gestation. Amino acid concentrations were determined using High-Performance Liquid Chromatography (HPLC), whereas fatty acids and volatile acids were analyzed by Gas Chromatography (GC) at each reproductive stage. The concentrations of both essential and non-essential amino acids declined significantly (P ≤ 0.0001) as pregnancy progressed, with the lowest levels observed on day 75 of gestation. Furthermore, saturated fatty acid levels were significantly higher (P ≤ 0.0001) during mid-gestation (day 75) compared to the other reproductive stages. In contrast, unsaturated fatty acid levels showed a declining trend toward the end of gestation. Additionally, volatile acid levels increased significantly (P ≤ 0.0001) up to mid-gestation (day 75). However, no significant associations were observed between amino acid concentrations and either litter size or offspring sex, except for asparagine, which was significantly higher (P ≤ 0.003) in ewes carrying male fetuses compared to those carrying females. The concentrations of fatty acids and volatile acids showed no significant association with either litter size or offspring sex. In conclusion, the blood profiles of amino acids, fatty acids, and volatile acids may serve as valuable biomarkers for pregnancy detection and for evaluating the nutritional requirements of the developing embryo across different reproductive stages. These findings have the potential to enhance reproductive management strategies and contribute to the improvement of sheep breeding programs.
References
Abadi, F. M., Mirfazeli, A., Zaeri, H., Nejabat, M., Taherizadeh, M., Ariaie, M., & Joshaghani, H. (2016). Analysis of plasma amino acids using RP-HPLC and pre-column derivatization with OPA/3-MPA. Medical Laboratory Journal, 10(2), 52–57. https://doi.org/10.18869/acadpub.mlj.10.2.52
Abdulkareem, T. A., Khalil, R. I., & Al-Ezzi, M. A. O. (2024). Effect of sperm freezability on some semen attributes and amino acid concentrations in seminal plasma of Holstein bulls. Iraqi Journal of Agricultural Sciences, 55(2), 665–674. https://doi.org/10.36103/pfgrpt81
Abdulkareem, T. A., Eidan, S. M., Al-Saidy, F. K., & Al-Hassani, N. K. (2023). Effect of pre-and post-mating vitamin AD3E treatment on reproductive performance of Awassi ewes. Iraqi Journal of Agricultural Sciences, 54(2), 431–437. https://doi.org/10.36103/ijas.v54i2.1717
Ahmadzadeh L., Hosseinkhani, A., Saedi, S., Daghigh Kia, H., Dadashi, M., & Jafarzadeh, J. (2016). Study of blood metabolites changes of purebred Ghezel and crossbred Arkhar Merinos × Ghezel ewes during late pregnancy. Iranian Journal of Animal Science Research, 8, 392–402. https://doi.org/10.22067/ijasr.v8i2.45117
Al-Gebouri, F. G., & Eidan, S. M. (2024a). Metabolites and semen characteristics in different bulls fertility. Iraqi Journal of Agricultural Sciences, 55(Special Issue), 206–216. https://doi.org/10.36103/ijas.v55iSpecial.1899
Al-Gebouri, F. G., & Eidan, S. M. (2024b). Effect of season and metabolites and semen traits of bulls. Iraqi Journal of Agricultural Sciences, 55(5), 1579–1587. https://doi.org/10.36103/193kb044
Ali, R. A. (2024). Impact of vitamin E and on the hematological, biochemical and productive parameters of pregnant Awassi ewes. International Journal of Life Science and Agriculture Research, 3(08), 709–716. https://doi.org/10.55677/ijlsar/V03I8Y2024-13
Ali, R. A., Habeeb, H. M. H., & Mahdi, A. K. (2024). Impact of short-term of estrous synchronization on some reproductive performance characteristics in Awassi sheep. Acta fytotechnica et Zootechnica, 27(3), 198–202. https://doi.org/10.15414/afz.2024.27.03.198-202
Alkass, J. E., Abdulkareem, T. A., & Al-Mjamei, S. M. (2004). Reproductive performance of Iraqi Awassi ewes in response to treatment with equine chorionic gonadotropin. Journal of Agricultural Investment, 2, 74–77.
Alon, T., Rosov, A., Lifshitz, L., & Moallem, U. (2023). Male fetuses negatively affect the vitality of the litter and the dam’s metabolic and physiological state in multifetal pregnant ewe. PLoS One, 18(5). https://doi.org/10.1371/journal.pone.0285338
Al-Saedi, A. J. A., & Abdulkareem, T. A. (2022). Comparison of semen quality for three lines of Holstein bulls. 1. Some immediate and microscopic characteristics. Iraqi Journal of Agricultural Sciences, 53(4), 752–759. https://doi.org/10.36103/ijas.v53i4.1585
Al-Thuwaini, T.M. (2022). Adiponectin and its physiological function in ruminant livestock. Reviews in Agricultural Science, 10, 115–122. https://doi.org/10.7831/ras.10.0_115
Aparicio, E., Martín-Grau, C., Hernández-Martinez, C., Voltas, N., Canals, J., & Arija, V. (2021). Changes in fatty acid levels (saturated, monounsaturated and polyunsaturated) during pregnancy. BMC Pregnancy and Childbirth, 21, 1–10. https://doi.org/10.1186/s12884-021-04251-0
Ashworth, C. J., Dwyer, C. M., McIlvaney, K., Werkman, M., & Rooke, J. A. (2011). Breed differences in fetal and placental development and fetomaternal amino acid status following nutrient restriction during early and mid-pregnancy in Scottish Blackface and Suffolk sheep. Reproduction Fertility and Development, 23(8), 1024–1033. https://doi.org/10.1071/RD10290
Bahr, A. H. M., Mahdi, A. S., & AL-Khuzai, H. M. (2022). Relationship of lactation and pregnancy stage with some hematological parameters of blood in Awassi ewes of different ages. Indian Journal of Ecology, 49(19), 249–251.
Bell, A. W., & Ehrhardt, R. A. (2002). Regulation of placental nutrient transport and implications for fetal growth. Nutrition Research Review, 15(02), 211–230. https://doi.org/10.1079/NRR200239
Bissonnette, J. M., Hohimer, A. R., & Chao, C. R. (1991). Unidirectional transport of glucose and lactate into brain of fetal sheep and guinea-pig. Experimental Physiology, 76(4), 515–523. https://doi.org/10.1113/expphysiol.1991.sp003517
Clarke, S. D. (2001). I. Molecular mechanism for polyunsaturated fatty acid regulation of gene transcription. American Journal of Physiology – Gastrointestinal and Liver Physiology, 281, G865–G869. https://doi.org/10.1152/ajpgi.2001.281.4.G865
De Boo, H. A., Van Zijl, P. L., Smith, D. E., Kulik, W., Lafeber, H. N., & Harding, J. E. (2005). Arginine and mixed amino acids increase protein accretion in the growth-restricted and normal ovine fetus by different mechanisms. Pediatric Research, 58(2), 270–277. https://doi.org/10.1203/01.PDR.0000169977.48609.55
Du, M., Ford, S. P., & Zhu, M. (2017). Optimizing livestock production efficiency through maternal nutritional management and fetal developmental programming. Animal Frontiers, 7(3), 5–11. https://doi.org/10.2527/af.2017-0122
Duggleby, S. L., & Jackson, A. A. (2002). Protein, amino acid and nitrogen metabolism during pregnancy: how might the mother meet the needs of her fetus? Current Opinion in Clinical Nutrition and Metabolic Care, 5, 503–509. https://doi.org/10.1097/00075197-200209000-00008
Duncan, D. B. (1955). Multiple ranges and multiple F tests. Biometrics, 11, 1–42.
Duttaroy, A. K. (2009). Transport of fatty acids across the human placenta: A review. Progress in Lipid Research, 48, 52–61. https://doi.org/10.1016/j.plipres.2008.11.001
Duttaroy, A. K., & Basak, S. (2022). Maternal fatty acid metabolism in pregnancy and Its consequences in the feto-placental development. Frontiers in Physiology, 12. https://doi.org/10.3389/fphys.2021.787848
Eidan, S. M., & Khudhir, S. A. (2023). Association between ATP1A1 gene polymorphisms with semen characteristics in Holstein bulls. Iraqi Journal of Agricultural Sciences, 54(2), 330–337. https://doi.org/10.36103/ijas.v54i2.1706
Eidan, S. M., Khalil, R. I., & Naser, A. F. (2024). Some of fatty acid and semen characteristics of Holstein bulls as influenced by different sperm freezability. Iraqi Journal of Agricultural Sciences, 55(2), 675–682. https://doi.org/10.36103/cg6xrf46
Erichsen, C., Heiser, A., Haack, N., Maclean, P., Dwyer, C. M., & McCoard, S. (2024). Increasing the understanding of nutrient transport capacity of the ovine placentome. Animals, 14(9), 1294. https://doi.org/10.3390/ani14091294
Edwards, A. K., McKnight, S. M., Askelson, K., McKnight, J. R., Dunlap, K. A., & Satterfield, M. C. (2020). Adaptive responses to maternal nutrient restriction alter placental transport in ewes. Placenta, 96, 1–9. https://doi.org/10.1016/j.placenta.2020.05.002
Flynn, N. E., Meininger, C. J., Haynes, T. E., & Wu, G. (2002). The metabolic basis of arginine nutrition and pharmacotherapy. Biomedicine and Pharmacotherapy, 56, 427–438. https://doi.org/10.1016/s0753-3322(02)00273-1
Freetly, H. C., & Ferrell, C. L. (1998). Net flux of glucose, lactate, volatile acids, and nitrogen metabolites across the portal-drained viscera and liver of pregnant ewes. Journal of Animal Science, 76(12), 3133–3145. https://doi.org/10.2527/1998.76123133x
Gao, H., Wu, G., Spencer, T. E., Johnson, G. A., Li, X., & Bazer, F. W. (2009). Select nutrients in the ovine uterine lumen. I. Amino acids, glucose, and ions in uterine luminal flushings of cyclic and pregnant ewes. Biology of Reproduction, 80, 86–93. https://doi.org/10.1095/biolreprod.108.071597
Goldansaz, S. A., Markus, S., Plastow, G., & Wishart, D. S. (2022). Predictive blood biomarkers of sheep pregnancy and litter size. Scientific Reports, 12, 10307. https://doi.org/10.1038/s41598-022-14141-w
Gulliver, C. E., Friend, M. A., King, B. J., Wilkins, J. F., & Clayton, E. H. (2013). A higher proportion of female lambs when ewes were fed oats and cottonseed meal prior to and following conception. Animal Production Science, 53(5), 464–471. https://doi.org/10.1071/AN12279
Habeeb, H. M. H., Aljebori, R. A. A., Mahdi, A., Hamza, H. F., & Kamil, B. R. (2023). Effect of Moringa oleifera leaf powder on Awassi ewe’s blood parameters. IOP Conference Series: Earth and Environmental Science, 1262(7). https://doi.org/10.1088/1755-1315/1262/7/072012
Ibrahim, F. K., Younis, S. T., Awaad, A. A., & Mageed, A. A. (2024). The Effect of ewe‘s age and the first birth on sex lamb, type of birth, litter size and the lamb‘s weight at birth and some genetic parameters. NTU Journal of Agriculture and Veterinary Science, 4(4), 235–240. https://doi.org/10.56286/ntujavs.v2i2
Innis, S. M. (2007). Fatty acids and early human development. Early Human Development, 83, 761–766. https://doi.org/10.1016/j.earlhumdev.2007.09.004
Kadhem, A. F., & Al-Thuwaini, T. M. (2022). Influence of litter size on the hematologic profile of Awassi ewes during gestation and lactation. Veterinary Integrative Sciences, 20(3), 625–633. https://doi.org/10.12982/VIS.2022.047
Khazaal, N. M., Alghetaa, H. F., & Al-Shuhaib, M. B. S. (2023). Hematological parameters as indicators for litter size and pregnancy stage in Awassi ewes. The Iraqi Journal of Veterinary Medicine, 47(1), 68–73. https://doi.org/10.30539/ijvm.v47i1.1504
Kwon, H., Spencer, T. E., Bazer, F. W., & Wu, G. (2003). Developmental changes of amino acids in ovine fetal fluids. Biology of Reproduction, 68(5), 1813–1820. https://doi.org/10.1095/biolreprod.102.012971
La, Y., Tang, J., Guo, X., Zhang, L., Gan, S., Zhang, X., & Chu, M. (2020). Proteomic analysis of sheep uterus reveals its role in prolificacy. Journal of Proteomics, 210. https://doi.org/10.1016/j.jprot.2019.103526
Luther, J. S., Windorski, E. J., Caton, J. S., Wu, G., Kirsch, J. D., Vonnahme, K. A., & Schauer, C. S. (2009). Effects of arginine supplementation on reproductive performance in Rambouillet ewes. Sheep Researches of Reproduction, 50, 11–13.
Majnooni, M. B., Mohammadi, B., Jalili, R., Babaei, A., & Bahrami, G. (2016). Determination of fatty acids by high-performance liquid chromatography and fluorescence detection using pre-column derivatization with 9-fluorenylmethyl chloroformate. Journal of Liquid Chromatography and Related Technologies, 39, 877–881. https://doi.org/10.1080/10826076.2016.1275000
Marconi, A. M., Battaglia, F. C., Meschia, G., & Sparks, J. W. (1989). A comparison of amino acid arteriovenous differences across the liver and placenta of the fetal lamb. American Journal of Physiology Endocrinology and Metabolism, 257(6), E909–E915. https://doi.org/10.1152/ajpendo.1989.257.6.E909
Marques, R. S., Cooke, R. F., Rodrigues, M. C., Brandão, A. P., Schubach, K. M., Lippolis, K. D., Moriel, P., Perry, G. A., Lock, A., & Bohnert, D. W. (2017). Effects of supplementing calcium salts of polyunsaturated fatty acids to late-gestating beef cows on performance and physiological responses of the offspring. Journal of Animal Science, 95, 5347–5357. https://doi.org/10.2527/jas2017.1606
McCoard, S. A., Sales, F. A., & Sciascia, Q. L. (2016). Amino acids in sheep production. Frontiers in Bioscience, Elite, 8, 264–288. https://doi.org/10.2741/E766
Mohammadi, V., Anassori, E., & Jafari, S. (2016). Measure of energy related biochemical metabolites changes during peri-partum period in Makouei breed sheep. Veterinary Research Forum, 7(1), 35–39.
Mohammed, S. H., & Al-Thuwaini, T. M. (2024). Investigation of litter size in Awassi Sheep and their correlation to adipose tissue biomarkers. Journal of Animal Health and Production, 12(4), 481–485. http://dx.doi.org/10.17582/journal.jahp/2024/12.4.481.485
Mohammed, M. H., Al-Thuwaini, T. M., & Al-Shuhaib, M. B. S. (2021). The association of the single – and twin-bearing with the lipid profile on the status of the reproductive hormones in Iraqi Awassi ewes. Advances in Animal and Veterinary Sciences, 9(9), 1456–1459. http://dx.doi.org/10.17582/journal.aavs/2021/9.9.1456.1459
Musa, K. S., & Abdulkareem, T. A. (2024). Some biochemical attributes in seminal plasma of Iraqi buffalo bulls and their relation to the semen quality. Iraqi Journal of Agricultural Sciences, 55(1), 402–412. https://doi.org/10.36103/nrfkex70
Nickles, K. R., Hamer, L., Coleman, D. N., & Relling, A. E. (2019). Supplementation with eicosapentaenoic and docosahexaenoic acids in late gestation in ewes changes adipose tissue gene expression in the ewe and growth and plasma concentration of ghrelin in the offspring. Journal of Animal Science, 97, 2631–2643. https://doi.org/10.1093/jas/skz141
Peñagaricano F., Wang, X., Rosa, G. J., Radunz, A. E., & Khatib, H. (2014). Maternal nutrition induces gene expression changes in fetal muscle and adipose tissues in sheep. BMC Genomics, 15, 1034. https://doi.org/10.1186/1471-2164-15-1034
Regnault, T. R. H., Friedman, J. E., Wilkening, R. B., Anthony, R. V., & Hay, W. W. (2005). Fetoplacental transport and utilization of amino acids in IUGR – A review. Placenta, 26, 52–62. https://doi.org/10.1016/j.placenta.2005.01.003
Roque-Jiménez, J. A., Rosa-Velázquez, M., Pinos-Rodríguez, J. M., Vicente-Martínez, J. G., Mendoza-Cervantes, G., Flores-Primo, A., Lee-Rangel, H. A., & Relling, A. E. (2021). Role of long chain fatty acids in developmental programming in Ruminants. Animals, 11, 762. https://doi.org/10.3390/ani11030762
SAS. (2018). Statistical Analysis System. User‘s Guide Statistical Version 9.1st ed. SAS Inst. Inc. Cary NC. USA.
Sauer, S. W., Okun, J. G., Hoffmann, G. F., Koelker, S., & Morath, M. A. (2008). Impact of short and medium-chain organic acids, acylcarnitines, and acyl-CoAs on mitochondrial energy metabolism. Biochimica et Biophysica Acta (BBA) – Bioenergetics, 1777(10), 1276–1282. https://doi.org/10.1016/j.bbabio.2008.05.447
Su, H. W., Yi, Y. C., Wei, T. Y., Chang, T. C., & Cheng, C. M. (2017). Detection of ovulation, a review of currently available methods. Bioengineering and Translational Medicine, 2(3), 238–246. https://doi.org/10.1002/btm2.10058
Tain, Y. L., & Hsu, C. N. (2024). Amino acids during pregnancy and offspring cardiovascular-kidney-metabolic health. Nutrients, 16(9), 1263. https://doi.org/10.3390/nu16091263
Thureen, P. J., Baron, K. A., Fennessey, P. V., & Hay, W. W. (2002). Ovine placental and fetal arginine metabolism at normal and increased maternal plasma arginine concentrations. Pediatric Research, 51(4), 464–471. https://doi.org/10.1203/00006450-200204000-00011
Ulwahhab, A. N. A., & R. I. Khalil. (2020). Effect of fetus sex, late gestation and lactation period of Awassi ewes on some hematological and biochemical parameters. Plant Archives, 20(1), 1645–1649.
Wang, J., Wu, Z., Li, D., Li, N., Dindot, S. V., Satterfield, M. C., Bazer, F. W., & Wu, G. (2012). Nutrition, epigenetics, and metabolic syndrome. Antioxidant and Redox Signaling, 17(2), 282–301. https://doi.org/10.1089/ars.2011.4381
Wang, X., Burghardt, R. C., Romero, J. J., Hansen, T. R., Wu, G., & Bazer, F. W. (2015). Functional roles of arginine during the peri-implantation period of pregnancy. III. Arginine stimulates proliferation and interferon tau production by ovine trophectoderm cells via nitric oxide and polyamine-TSC2-MTOR signaling pathways. Biology of Reproduction, 92, 1–17. https://doi.org/10.1095/biolreprod.114.125989
Wooldridge, A. L., Bischof, R. J., Liu, H., Heinemann, G. K., Hunter, D. S., Giles, L. C., & Gatford, K. L. (2018). Late-gestation maternal dietary methyl donor and cofactor supplementation in sheep partially reverses protection against allergic sensitization by IUGR. American Journal of Physiology Regulation, Integrative and Comparative Physiology, 314(1), R22–R33. https://doi.org/10.1152/ajpregu.00549.2016
Wu, G., Bazer, F. W., Cudd, T. A., Meininger, C. J., & Spencer, T. E. (2004). Maternal nutrition and fetal development. Journal of Nutrition, 134, 2169–2172. https://doi.org/10.1093/jn/134.9.2169
Zhai, Y., Xia, F., Shi, L., Ma, W., Lv, X., Sun, W., Ji, P., Gao, S., Machaty, Z., & Liu, G. (2023). Early pregnancy markers in the serum of ewes identified via proteomic and metabolomic analyses. International Journal of Molecular Sciences, 24, 14054. https://doi.org/10.3390/ijms241814054
Ziętek, M., Celewicz, Z., & Szczuko, M. (2021). Short-chain fatty acids, maternal microbiota and metabolism in pregnancy. Nutrients, 13(4), 1244. https://doi.org/10.3390/nu13041244
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Talal A. Abdulkareem, Rwaida A. Ali
This work is licensed under a Creative Commons Attribution 4.0 International License.
