Effect of in ovo Injection of Royal Jelly on Post-Hatch Growth Performance and Immune Response in Broiler Chickens Challenged with Newcastle Disease Virus

Document Type : Research Article


1 Faculty of Animal Science, Gorgan University of Agricultural Science and Natural Resource, Gorgan, Iran

2 Department of Animal Science, Shiraz University, Shiraz, Iran

3 Department of Animal Science, Islamic Azad University, Qaemshahr, Iran

4 Faculty of Food Scienceand Technology, Gorgan University of Agricultural Science and Natural Resource, Gorgan, Iran

5 Department of Microbiology and Immunology, Gorgan University of Medical Science, Gorgan, Iran


The aim of this experiment was to study the effect of in ovoinjection of royal jelly (RJ) on growth performance and immune response in broiler chickens challenged with Newcastle disease virus. A total of 200 hatching eggs were divided into four groups of 50 eggs each one using a completely randomized design. The eggs were injected with sterilized sodium bicarbonate buffer (pH=10.4) or RJ (88%) in a 0.3 mL volume into the albumen at d 7 of incubation. A non-injected control and a dry punch control (shell pricked without injecting solution; needle) were included. Chicks hatched from the respective treatment group were reared in floor pens until d 28. The chickens were challenged against Newcastle disease virus on days 7 and 21. On d 21, the chickens emanatedfrom the RJ injected egg had the highest body weight. In ovo injection of RJ had a significant effect on feed intake in broiler chickens during starter period (d 1 to 21) and throughout the trial (d 1 to 28; P<0.05). On d 14, serum antibody titer against Newcastle disease virus (NDV) was not significantly different among treatments. On d 14 and 28, heterophil and lymphocyte number and their ratio were affected by in ovo injection. In conclusion, the results of the present study suggested that in ovo injection of RJ on d 7 of incubation exerted a beneficial effect on growth in starter phase and could stimulate feed intakein broiler chickenschallenged with Newcastle disease virus, although the antibody titer against NDV was notinfluenced.


  1. Allan W.H. and Gough R.E. (1974). A standard haemagglutination inhibition test for Newcastle disease. A comparison of macro and micro methods. Vet. Rec. 95, 120-123.
    Baker D.H. (1994). Ideal amino acid profile for maximal protein secretion and minimal nitrogen excretion in swine and poultry. Pp. 134-139 in Proc. Cornell Nutr. Conf., Ithaca, NY.
    Bhanja S.K., Mandal A.B. and Goswami T.K. (2004). Effect of in ovo injection of amino acids on growth, immune response, development of digestive organs and carcass yields of broilers. Indian J. Poult. Sci. 39, 212-218.
    Bhanja S.K. and Mandal A.B. (2005). Effect of in ovo injection of critical amino acids on pre- and post-hatch growth, immunocompetence and development of digestive organs in broiler chickens. Asian Aus. J. Anim. Sci. 18, 524-531.
    Bilikova K., Hanes J., Nordhoff E., Saenger W., Klaudiny J. and Simuth J. (2002). Apisimin, a new serine-valine-rich peptide from honeybee (Apis mellifera) royal jelly: purification and molecular characterization. Febs Lett. 528, 125-129.
    Bilikova K., Wu G. and Simuth J. (2001). Isolation of a peptide fraction from honeybee royal jelly as a potential antifoulbrood factor. Apidol. 32, 275-283.
    Bonomi A., Bonomi B.M. and Quarantelli A. (2001). Royal jelly in turkey feeding; Royal jelly in guinea-fowlfeeding. Riv. Sci. Anim. 30, 49-60.
    Chauvin R. (1968). Action physiologique et therapeutique des produits de la ruche. Traite de biologie de l'abeille. Masson. Paris. 13, 116-154.
    Cheema M.A., Qureshi M.A. and Havenstein G.B. (2003). A comparison of the immune response of a 2001 commercial broiler with a 1957 random bred broiler strain when fed representative 1957 and 2001 broiler diets. Poult. Sci. 82, 1519-1529.
    Cheison S.C. and Wang Z. (2003). Bioactive milk peptides: Redefining the food-drug interphase. A review. Afr. J. Food Nutr. Sci. Agric. Dev.3, 29-38.
    Emmerson D.A. (1997). Commercial approaches to genetic selection for growth and feed conversion in domestic poultry. Poult. Sci. 76, 1121-1125.
    Foye O.T., Uni Z. and Ferket P.R. (2006). Effect of in ovo feeding egg white protein, beta-hydroxy-beta-methylbutyrate and carbohydrates on glycogen status and neonatal growth of turkeys. Poult. Sci. 85, 1185-1192.
    Freeman B.M. (1987). The stress syndrome. World’s Poult. Sci. J. 43, 15-19.
    Fujii A. (1995). Pharmacological effects of royal jelly. Honeybee Sci. 16, 97-104.
    Fujiwara S., Imai J., Fujiwara M., Yaeshima T., Kawashima T. and Kobayashi K. (1990). A potent antibacterial protein in royal jelly. Purification and determination of the primary structure of royalisin. J. Biol. Chem. 265, 11333-11337.
    Gonnard P. and N'guyen C.C. (1957). Action of royal jelly on oxygen consumption in tissues in vitro. Ann. Pharmacol. Fr. 15, 383-393.
    Gore A.B. and Qureshi M.A. (1997). Enhancement of humoral and cellular immunity by vitamin E after embryonic exposure. Poult. Sci. 76, 984-991.
    Gross W.B. and Siegel H.S. (1983). Evaluation of the heterophil/lymphocyte ratio as a measure of stress in chickens. Avian Dis. 27, 972-979.
    Hanes J. and Simúth J. (1992). Identification and partial characterization of the major royal jelly protein of the honey bee (Apis mellifera). J. Apic. Res. 31, 22-26.
    Havenstein G.B., Ferket P.R., Scheideler S.E. and Larson B.T. (1994). Growth, livability and feed conversion of 1957 vs. 1991 broilers when fed 'typical' 1957 and 1991 broiler. Poult. Sci. 73, 1785-1794.
    Henry M.H. and Burke W.H. (1999). The effects of in ovo administration of testosterone or an antiandrogen on growth of chick embryos and embryonic muscle characteristics. Poult. Sci. 78, 1006-1013.
    Huff G.R., Huff W.E., Balog J.M., Rath N.C., Anthony N.B. and Nestor K.E. (2005). Stress response differences and disease susceptibility reflected by heterophil to lymphocyte ratio in turkeys selected for increased body weight.Poult. Sci. 84, 709-717.
    Jochemsen P. and Jeurissen S.H. (2002). The localization and uptake of in ovo injected soluble and particulate substances in the chicken. Poult. Sci. 81, 1811-1817.
    Johnston P.A., Liu H., O’Connell T., Phelps P., Bland M., Tyczkowski J., Kemper A., Harding T., Avakian A., Haddad E., Whitfill C., Gildersleeve R. and Ricks C.A. (1997). Applications in in ovo technology. Poult. Sci. 76, 165-178.
    Kadam M.M., Bhanja S.K., Mandal A.B., Thakur R., Vasan P., Bhattacharyya A. and Tyagi J.S. (2008). Effect of in ovo threonine supplementation on early growth, immunological responses and digestive enzyme activities in broiler chickens. Br. Poult. Sci. 49, 736-741.
    Kamakura M. (2011). Royalactin induces queen differentiation in honeybees. Nature. 473, 478-483.
    Keralapurath M.M., Corzo A., Pulikanti R., Zhai W. and Peebles E.D. (2010a). Effects of in ovo injection of L-carnitine on hatchability and subsequent broiler performance and slaughter yield. Poult. Sci. 89, 1497-1501.
    Keralapurath M.M., Keirs R.W., Corzo A., Bennett L.W., Pulikanti R. and Peebles E.D. (2010b). Effects of in ovo injection of L-carnitine on subsequent broiler chick tissue nutrient profiles. Poult. Sci. 89, 335-341.
    Kidd M.T., Kerr B.J. and Anthony N.B. (1997). Dietary interactions between lysine and threonine in broilers. Poult. Sci. 76, 608-614.
    Kocamis H., Yeni Y.N., Kirkpatrick-Keller D.C. and Killefer J. (1999). Postnatal growth of broilers in response to in ovo administration of chicken growth hormone. Poult. Sci.

    1. 78, 1219-1226.

    Kowalski A., Mormede P., Jakubowski K. and Jedlinska-Krakowska M. (2002). Comparison of susceptibility to stress in two genetic lines of turkey broilers BUT-9 and Big-6. Pol. J. Vet. Sci. 5, 145-150.
    Krylov V. and Sokolskii C. (2000). Royal jelly (in Russian). Agroprompoligrafist Krasnodar. Pp. 214.
    Liu X.Y. (1999). Stress and immunity. Pp. 230-252 in Poultry Immunology. Yin, T.B. Ed. China Agriculture Press, Beijing, China.
    Maroufyan E., Kasim A., Hashemi S.R., Loh T.C., Hair Bejo M. and Davoodi H. (2010a). The effect of methionine and threonine supplementations on immune responses of broiler chickens challenged with infectious bursal disease. Am. J. Appl. Sci. 7, 44-50.
    Maroufyan E., Kasim A., Hashemi S.R., Loh T.C. and Hair Bejo M. (2010b). Responses of performance and differential leukocyte count to methionine and threonine supplementations on broiler chickens challenged with infectious bursal disease intropical condition. Asian J. Biol. Sci. 3, 68-76.
    Maxwell M.A. (1993). Avian blood leucocyte responses to stress. World’s Poult. Sci. J. 39, 34-43.
    NRC. (1994). Nutrient requirements of poultry. 9th rev. Ed. National Academy Press, Washington D.C. USA.
    Ohta Y., Tsushima N., Koide K., Kidd M.T. and Ishibashi T. (1999). Effect of amino acid injection in broiler breeder eggs on embryonic growth and hatchability of chicks. Poult. Sci. 78, 1493-1498.
    Oselli E., Caboni M.F., Sabatini A.G., Marcazzan G.L. and Lercker G. (2003). Determination and changes of free amino acids in royal jelly during storage. Apidol. 34,129-137.
    SAS Institute. (2005). SAS Proprietary Software Release 9.1. SAS Inst. Inc., Cary, NC.
    Serra-Bonvehi J. (1990). Studies on the proteins and free amino acids of royal jelly. Anal. Bromatol. 42,353-365.
    Siegel H.S. (1985). Immunological responses as indicators of stress. World’s Poult. Sci. J. 41, 36-44.
    Simuth J., Bilikova K. and Kovacora E. (2003). Royal Jelly proteins as a tool for development of functional ingredients for health.Pp. 312-340 in Proc. 38th Congress of Apimondia, Ljubljana, Slovenia.
    Tako E., Ferket P.R. and Uni Z. (2004). Effects of in ovo feeding of carbohydrates and β-hydroxy-β-methylbutyrate on the development of chicken intestine. Poult. Sci. 83, 2023-2028.
    Uni Z., Ferket P.R., Tako E. and Kedar O. (2005). In ovo feeding improves energy status of late-term chicken embryos. Poult. Sci. 84, 764-770.
    Zhai W., Neuman S., Latour M.A. and Hester P.Y. (2008). The effect of in ovo injection of L-carnitine on hatchability of White Leghorns. Poult. Sci. 87, 569-572.