Physico-chemical properties of late-incubation egg amniotic fluid and a potential in ovo feed supplement

  • Omede, A.A. (School of Environmental and Rural Science, University of New England) ;
  • Bhuiyan, M.M. (School of Environmental and Rural Science, University of New England) ;
  • lslam, A.F. (School of Environmental and Rural Science, University of New England) ;
  • Iji, P.A. (School of Environmental and Rural Science, University of New England)
  • Received : 2016.09.07
  • Accepted : 2017.01.25
  • Published : 2017.08.01


Objective: This study explored the physico-chemical properties of late-incubation egg amniotic fluid and a potential in ovo feed (IOF) supplement. Methods: Amniotic fluid was collected from broiler breeders (Ross 308, 51 weeks and Cobb 500, 35 weeks) on day 17 after incubation. A mixture of high-quality soy protein supplement - Hamlet Protein AviStart (HPA) was serially diluted in MilliQ water to obtain solutions ranging from 150 to 9.375 mg/mL. The mixtures were heat-treated (0, 30, 60 minutes) in a waterbath ($80^{\circ}C$) and then centrifuged to obtain supernatants. The amniotic fluid and HPA supernatants were analysed for their physico-chemical properties. Results: Only viscosity and $K^+$ were significantly (p<0.05) different in both strains. Of all essential amino acids, leucine and lysine were in the highest concentration in both strains. The osmolality, viscosity and $pCO_2$ of the supernatants decreased (p<0.05) with decreasing HPA concentration. Heat treatment significantly (p<0.05) affected osmolality, pH, and $pCO_2$, of the supernatants. The interactions between HPA concentration and heat treatment were significant with regards to osmolality (p<0.01), pH (p<0.01), $pCO_2$ (p<0.05), glucose (p<0.05), lactate (p<0.01) and acid-base status (p<0.01) of HPA solutions. The $Ca^{2+}$, $K^+$, glucose, and lactate increased with increasing concentration of HPA solution. The protein content of HPA solutions decreased (p<0.05) with reduced HPA solution concentrations. The supernatant from 150 mg/mL HPA solution was richest in glutamic acid, aspartic acid, arginine and lysine. Amino acids concentrations were reduced (p<0.05) with each serial dilution but increased with longer heating. Conclusion: The values obtained in the primary solution (highest concentration) are close to the profiles of high-protein ingredients. This supplement, as a solution, hence, may be suitable for use as an IOF supplement and should be tested for this potential.


Amino Acids;Amniotic Fluids;In ovo;Physico-chemical Properties;Protein


  1. Kadam MM, Bhanja SK, Mandal AB, et al. Effect of in ovo threonine supplementation on early growth, immunological responses and digestive enzyme activities in broiler chickens. Br Poult Sci 2008;49:736-41.
  2. Tako E, Ferket PR, Uni Z. Effects of in ovo feeding of carbohydrates and ${\beta}$-Hydroxy-${\beta}$-Methylbutyrate on the development of chicken intestine. Poult Sci 2004;83:2023-8.
  3. Gore AB, Qureshi MA. Enhancement of humoral and cellular immunity by vitamin E after embryonic exposure. Poult Sci 1997;76:984-91.
  4. Dalloul RA, Lillehoj HS, Klinman DM, et al. In ovo administration of CpG oligodeoxynucleotides and the recombinant microneme protein MIC2 protects against Eimeria infections. Vac 2005;23:3108-13.
  5. Keralapurath MM, Corzo A, Pulikanti R, Zhai W, Peebles ED. Effects of in ovo injection of L-carnitine on hatchability and subsequent broiler performance and slaughter yield. Poult Sci 2010;89:1497-501.
  6. Kocamis H, Yeni YN, Brown CU, et al. Effect of in ovo administration of insulin-like growth factor-I on composition and mechanical properties of chicken bone. Poult Sci 2000;79:1345-50.
  7. McGruder BM, Zhai W, Keralapurath MM, et al. Effects of in ovo injection of electrolyte solutions on the pre- and posthatch physiological characteristics of broilers. Poult Sci 2011;90:1058-66.
  8. Kadam MM, Barekatain MR, Bhanja SK, Iji PA. Prospects of in ovo feeding and nutrient supplementation for poultry: the science and commercial applications - a review. J Sci Food Agric 2013;93:3654-61.
  9. Salmanzadeh M. Effects of in ovo injection of glucose on hatchability, hatching weight and subsequent performance of newly-hatched chicks. Braz J Poult Sci 2012;14:137-40.
  10. Ohta Y, Kidd MT. Optimum site for in ovo amino acid injection in broiler breeder eggs. Poult Sci 2001;80:1425-9.
  11. Baggott GK. Development of extra-embryonic membranes and fluid compartments. In: Deeming DC, editor. Perspectives in fertilisation and embryonic development in poultry. Lincolnshire, UK: Ratite Conference Books; 2001.
  12. Romanoff AL, Romanoff AJ. Biochemistry of the avian embryo: a quantitative analysis of prenatal development. New York: Interscience publishers, A division of John Wiley and Sons; 1967.
  13. Uni Z, Ferket PR. Enhancement of development of oviparous species by in ovo feeding. ed. U. S. Patent. USA: North Carolina State University, Raleigh, NC (US); Yissum Research Development Company of University of Jerusalem, Jerusalem (IL); 2003. Patent No: US6592878B2.
  14. de Oliveira JE. Effects of in ovo feeding on turkey embryos development, energy status, intestinal maturation, gene expression and posthatch development. PhD thesis, Raleigh, NC: North Carolina State University; 2007.
  15. Kemps BJ, Bamelis FR, Mertens K, et al. The assessment of viscosity measurements on the albumen of consumption eggs as an indicator for freshness. Poult Sci 2010;89:2699-703.
  16. Minitab Inc. $^Minitab{(R)}$ Statistical Packge. State College PA, USA: Minitab Inc.; 2013.
  17. Tona KO, De Ketelaere B, Decuypere E, Bruggeman V. Effects of age of broiler breeders and egg storage on egg quality hatchability, chick quality, chick weight, and chick posthatch growth to forty-two days. J Appl Poult Res 2004;13:10-8.
  18. Wolanski NJ, Renema RA, Robinson FE, Carney VL, Fancher BI. Relationships among egg characteristics, chick measurements, and early growth traits in ten broiler breeder strains. Poult Sci 2007;86:1784-92.
  19. Hoyt DF. Osmoregulation by avian embryos: The allantois functions like a toad's bladder. Physiol Zool 1979;52:354-62.
  20. Bolin GM. Incubation humidity as an environmental stressor on the osmoregulatory developmental program of the chicken, Gallus gallus domesticus. PhD thesis. North Texas, TX: University of North Texas; 2009.
  21. Carinci P, Manzoli-Guidott L. Albumen absorption during chick embryogenesis. J Embryol Exp Morph 1968;20:107-18.
  22. Ferket P, de Oliveira J, Ghane A, Uni Z. Effect of in ovo feeding solution osmolality on hatching turkeys. Poult Sci 2005;84(Supplement 1):118-9.
  23. Kornasio R, Halevy O, Kedar O, Uni Z. Effect of in ovo feeding and its interaction with timing of first feed on glycogen reserves, muscle growth, and body weight. Poult Sci 2011;90:1467-77.
  24. Gaafar KM, Selim SA, El-Ballal SS. Effect of in-ovo administration with two levels of amino acids mixture on the performance of Muscovy ducks. Emir J Food Agric 2013;25:58-65.
  25. Sigwart JD, Lyons G, Fink A, et al. Elevated p$CO_2$ drives lower growth and yet increased calcification in the early life history of the cuttlefish Sepia officinalis (Mollusca: Cephalopoda). ICES J Mar Sci 2016;73:970-80.
  26. Molenaar R, van den Borne JJ, Hazejager E, et al. High environmental temperature increases glucose requirement in the developing chicken embryo. PLoS One, 2013;8,e59637.
  27. Molenaar R, van den Anker I, Meijerhof R, Kemp B, van den Brand H. Effect of eggshell temperature and oxygen concentration during incubation on the developmental and physiological status of broiler hatchlings in the perinatal period. Poult Sci 2011;90:1257-66.
  28. Batal AB, Dale NM, Saha UK. Mineral composition of corn and soybean meal. J Appl Poult Res 2010;19:361-4.
  29. Uni Z, Ferket PR. Methods for early nutrition and their potential. World's Poult Sci J 2004;60:101-11.
  30. Shafey TM, Mahmoud AH, Alsobayel AA, Abouheif MA. Effects of in ovo administration of amino acids on hatchability and performance of meat chickens. South Afric J Anim Sci 2014;44:123-30.
  31. Wu G, Bazer FW, Cudd TA, Meininger CJ, Spencer TE. Maternal Nutrition and Fetal development. J Nutr 2004;134:2169-72.
  32. Newsholme EA, Calder PC. The proposed role of glutamine in some cells of the immune system and speculative consequences for the whole animal. Nutrition 1997;13:728-30.
  33. Al-Murrani WK. Effect of injecting amino acids into the egg on embryonic and subsequent growth in the domestic fowl. Br Poult Sci 1982;23:171-4.
  34. Ohta Y, Kidd MT, Ishibashi T. Embryo growth and amino acid concentration profiles of broiler breeder eggs, embryos, and chicks after in ovo administration of amino acids. Poult Sci 2001;80:1430-6.
  35. Li P, Yin YL, Li D, Kim SW, Wu G. Amino acids and immune function. Br J Nutr 2007;98:237-52.
  36. Murray KR, Granner DK, Rodwell VW. Metabolism of proteins and amino acids. In: Harper's Illustrated Biochemistry, New York, USA:McGraw Hill; 2006.
  37. Wu G, Bazer FW, Burghardt RC et al. Proline and hydroxyproline metabolism: implications for animal and human nutrition. Amino Acids 2011;40:1053-63.
  38. Ohta Y, Tsushima N, Koide K, Kidd MT, Ishibashi T. Effect of amino acid injection in broiler breeder eggs on embryonic growth and hatchability of chicks. Poult Sci 1999;78:1493-8.