A.H., B., A.M., A., N.M., E. (2025). Effect of Dietary Bacillus subtilis Supplementation on Lymphoid Organ Weights and Antibody Production Against the Newcastle Disease Vaccine in Broiler Chicks. Journal of Applied Veterinary Sciences, 10(2), 64-74. doi: 10.21608/javs.2025.352341.1513
Bashir A.H.; Abdelatif A.M.; Elkhair N.M.. "Effect of Dietary Bacillus subtilis Supplementation on Lymphoid Organ Weights and Antibody Production Against the Newcastle Disease Vaccine in Broiler Chicks". Journal of Applied Veterinary Sciences, 10, 2, 2025, 64-74. doi: 10.21608/javs.2025.352341.1513
A.H., B., A.M., A., N.M., E. (2025). 'Effect of Dietary Bacillus subtilis Supplementation on Lymphoid Organ Weights and Antibody Production Against the Newcastle Disease Vaccine in Broiler Chicks', Journal of Applied Veterinary Sciences, 10(2), pp. 64-74. doi: 10.21608/javs.2025.352341.1513
A.H., B., A.M., A., N.M., E. Effect of Dietary Bacillus subtilis Supplementation on Lymphoid Organ Weights and Antibody Production Against the Newcastle Disease Vaccine in Broiler Chicks. Journal of Applied Veterinary Sciences, 2025; 10(2): 64-74. doi: 10.21608/javs.2025.352341.1513
Effect of Dietary Bacillus subtilis Supplementation on Lymphoid Organ Weights and Antibody Production Against the Newcastle Disease Vaccine in Broiler Chicks
1Department of Physiology, Faculty of Veterinary Medicine, El-Salam University, Sudan
22Department of Physiology, Faculty of Veterinary Medicine, University of Khartoum, Sudan
3Department of Biomedical Sciences, College of Veterinary Medicine, King Faisal University, Al-Ahsa, 31982, Saudi Arabia Department of Physiology, Faculty of Veterinary Medicine, University of Khartoum, 13314, Shambat, Sudan
Receive Date: 12 January 2025,
Revise Date: 19 February 2025,
Accept Date: 01 March 2025
Abstract
This study investigated the impact of Bacillus subtilis supplementation on lymphoid organ weights and antibody production in response to the Newcastle Disease Vaccine (NDV) in Ross 308 chicks. A total of 120 one-day-old chicks were divided into four groups with three replicates of 10 birds each, receiving different concentrations of Bacillus subtilis (Bs): Control (Bs-0), 0.05% (Bs-0.05), 0.10% (Bs-0.1), and 0.15% (Bs-0.15) for six weeks. Newcastle disease, infectious bronchitis and Gumboro disease vaccines were administered in drinking water on days 3, 11 and 15 of age, respectively. Blood and serum samples were harvested on day 21 to assess antibody production against NDV and lymphoid organ weights on day 42. On day 21, the antibody titers against the NDV vaccine were significantly higher (P≤0.05) in all chick groups supplemented with Bacillus subtilis compared to the control; the highest titer was observed in the Bs-0.15 group. By day 42, the final body weight (FBW) was significantly (P≤0.01) higher in the supplemented group Bs-0.15 than in others. The supplemented group, Bs-0.05, showed a numerically higher FBW than Bs-0.1 and the control group. The absolute and relative weights of lymphoid organs (spleen, thymus and bursa of Fabricius) were significantly higher (P≤0.01) in the supplemented groups than in the control. The highest organ weights were recorded in chicks receiving the highest concentration of Bacillus subtilis (Bs-0.15). These findings demonstrate that dietary supplementation with Bacillus subtilis at 0.15% enhances BW and the immune response and organ development associated with immunity in chicks. This supplementation levelimproves immune system health, bolsters disease resistance, and inspires and motivates further exploration of its potential utility in poultry nutrition strategies.
ABREU, R., SEMEDO-LEMSADDEK, T., CUNHA, E., TAVARES, L., and OLIVEIRA, M., 2023. Antimicrobial Drug Resistance in Poultry Production: Current Status and Innovative Strategies for Bacterial Control. Microorganisms 11(4), 953. https://doi.org/10.3390/microorganisms11040953
AHMED, S., and SIDDIQUI, S.M., 2021. Effect of dietary probiotics on immune modulation and NDV antibody titers in broiler chickens. Journal of Applied Poultry Research 30(4), 100210. https://doi.org/10.1016/j.japr.2021.100210
ALEXANDER, D.J. 2000. Newcastle disease and other avian paramyxoviruses. Revue Scientifique et Technique de l'OIE 19(2), 443–462. https://doi.org/10.20506/rst.19.2.1236
APATA, D.F. 2008. Growth performance, nutrient digestibility, and immune response of broiler chicks fed diets supplemented with a probiotic. International Journal of Poultry Science 7(11), 1056–1061. https://doi.org/10.1002/jsfa.3214
ARIF, M., AKTERUZZAMAN, M., ISLAM, S.S., DAS, B.C., SIDDIQUE, M.P., and LUTFUL KABIR, S.M., 2021. Dietary supplementation of Bacillus-based probiotics on the growth performance, gut morphology, intestinal microbiota and immune response in low biosecurity broiler chickens. Veterinary and Animal Science 14, 100216. https://doi.org/10.1016/j.vas.2021.100216
AWAD, W.A., GHAREEB, K., ABDEL-RAHEEM, S., and BÖHM, J., 2009. Effect of dietary inclusion of probiotic and symbiotic on broiler chickens' growth performance, organ weights, and intestinal histomorphology. Poultry Science 88, 49–55. https://doi.org/10.3382/ps.2008-00244
AYANA, G.U., and KAMUTAMBUKO, R., 2024. Probiotics in Disease Management for Sustainable Poultry Production. Advanced Gut & Microbiome Research 2024(1), 4326438. https://doi.org/10.1155/2024/4326438
CECCOPIERI, C., and MADEJ, J.P., 2024. Chicken Secondary Lymphoid Tissues-Structure and Relevance in Immunological Research. Animals14(16), 2439. https://doi.org/10.3390/ani14162439
CHANDRASEKARAN, P., WEISKIRCHEN, S., and WEISKIRCHEN, R., 2024. Effects of Probiotics on Gut Microbiota: An Overview. International Journal of Molecular Sciences 25(11), 6022. https://doi.org/10.3390/ijms25116022
CHENG, J., LEI, H., XIE, C., CHEN, J., YI, X., ZHAO, F., YUAN, Y., CHEN, P., HE, J., LUO, C., SHU, D., QU, H., and JI, J., 2023. B Lymphocyte Development in the Bursa of Fabricius of Young Broilers is Influenced by the Gut Microbiota. Microbiology Spectrum 11(2), e04799–22. https://doi.org/10.1128/spectrum.04799-22
CHI, H., PEPPER, M., and THOMAS, P.G., 2024. Principles and therapeutic applications of adaptive immunity. Cell 187(9), 2052–2078. https://doi.org/10.1016/j.cell.2024.03.037
CIURESCU, G., DUMITRU, M., GHEORGHE, A., UNTEA, A., and DRĂGHICI, R., 2020. Effect of Bacillus subtilis on growth performance, bone mineralization, and bacterial population of broilers fed with different protein sources. Poultry Science 99(11), 5960. https://doi.org/10.1016/j.psj.2020.08.075
COOPER, M.D., PETERSON, R.D.A., and GOOD, R.A., 1966. Delineation of the thymus-dependent and bursa-dependent systems of the chicken. Journal of Experimental Medicine 123(1), 75–102. https://doi.org/10.1084/jem.123.1.75
DIMITROV, K.M., AFONSO, C.L., YU, Q., and MILLER, P.J., 2016. Newcastle disease vaccines—A solved problem or a continuous challenge? Veterinary Microbiology 206, 126–136. https://doi.org/10.1016/j.vetmic.2016.12.019
DONG, Y., LI, R., LIU, Y., MA, L., ZHA, J., QIAO, X., CHAI, T., and WU. B., 2020. Benefit of Dietary Supplementation with Bacillus subtilis BYS2 on Growth Performance, Immune Response, and Disease Resistance of Broilers. Probiotics and antimicrobial proteins 12, 1385–1397 (2020). https://doi.org/10.1007/s12602-020-09643-w
EL-SAYED, Y., KHALIL, W., FAYEZ, N., and MOHAMED ABDEL-FATTAH, A.F., 2024. Enhancing effect of oregano essential oil and Bacillus subtilis on broiler immune function, intestinal morphology and growth performance. BMC Veterinary Research20, 112. https://doi.org/10.1186/s12917-024-03960-w
ERGÜN, D., TARTAR, G., and YAZGAN-KARATAŞ, A., 2024.B. subtilis Probiotics for Humans, Animals and Plants: Mechanisms, Applications and Prospects. Bacillus subtilis- Functionalities and Emerging Applications. IntechOpen. http://dx.doi.org/10.5772/intechopen.1007721
GAO, Z., WU, H., SHI, L., ZHANG, X., SHENG, R., YIN, F., and GOONERATNE, R., 2017. Study of Bacillus subtilis on growth performance, nutrition metabolism and intestinal microflora of 1 to 42 d broiler chickens. Animal Nutrition 3(2), 109. https://doi.org/10.1016/j.aninu.2017.02.002
Glick, B. 1991. Historical perspective: the bursa of Fabricius and its influence on B-cell development, past and present. Veterinary immunology and immunopathology 30(1), 3–12. https://doi.org/10.1016/0165-2427(91)90003-u
HATAB M.H., ELSAYED M.A., and IBRAHIM N.S., 2016. Effect of some biological supplementation on productive performance, physiological and immunological response of layer chicks. Journal of Radiation Research and Applied Sciences 9, 185–192. https://doi.org/10.1016/j.jrras.2015.12.008
JEONG, J.S., and KIM, I.H., 2014. Effect of Bacillus subtilis UBT-MO2 as a probiotic in broilers. Poultry Science 93(8), 2118–2125. https://doi.org/10.3382/ps.2014-03916
KHONGTHONG, S., PIEWNGAM, P., ROEKNGAM, N., MALIWAN, P., KONGPUCKDEE, S., JEENKEAWPLEAM, J., and RODJAN, P., 2025. Effects of dietary Bacillus subtilis 14823 on growth performance, gut barrier integrity and inflammatory response of broilers raised in a stressful tropical environment. Poultry Science 104(1), 104518. https://doi.org/10.1016/j.psj.2024.104518
KELLY, L.M., and ALWORTH, L.C., 2013. Techniques for collecting blood from the domestic chicken. Laboratory Animal 42(10), 359–361. https://doi.org/10.1038/laban.394
KIARIE, E., ROMERO, L.F., and NYACHOTI, C.M., 2022. Probiotics and postbiotics in poultry production: Effects on gut health and NDV vaccine response. Animal Feed Science and Technology 287, 115277. https://doi.org/10.1016/j.anifeedsci.2021.115277
LARSBERG, F., SPRECHERT, M., HESSE, D., LOH, G., BROCKMANN, G. A., and KREUZER-REDMER, S., 2023. Probiotic Bacillus Strains Enhance T Cell Responses in Chicken. Microorganisms 11(2), 269. https://doi.org/10.3390/microorganisms11020269
LEE, K.W., LEE, S.H., LILLEHOJ, H.S., LI, G.X., JANG, S.I., and BABU, U.S., 2020. Effects of direct-fed microbials on growth performance, gut microbiota, and immune responses in broiler chickens. Poultry Science 99(4), 2454–2460. https://doi.org/10.1016/j.psj.2019.10.012
LI, D., FANG, S., HE, F., FAN, X., WANG, T., CHEN, Z., and WANG, M., 2024. Postbiotic derived from Bacillus subtilis ACCC 11025 improves growth performance, mortality rate, immunity, and tibia health in broiler chicks. Frontiers in Veterinary Science 11, 1414767. https://doi.org/10.3389/fvets.2024.1414767
LI, W., ZHANG, H., and ZHANG, Z., 2023.Bacillus subtilis and its role in enhancing Newcastle Disease vaccine efficacy in poultry: A review. Poultry Science 102(3), 101472. https://doi.org/10.1016/j.psj.2023.101472
LILLEHOJ, H.S. 1991. Cell-mediated immunity in parasitic and bacterial diseases of poultry. Poultry Science 70(2), 1167–1175.
MA, Y., WANG, W., ZHANG, H., WANG, J., ZHANG, W., GAO, J., WU, S., and QI, G., 2018. Supplemental Bacillus subtilis DSM 32315 manipulates broiler chickens' intestinal structure and microbial composition. Scientific Reports 8(1), 1–13. https://doi.org/10.1038/s41598-018-33762-8
MAYO, M.A. 2002. A summary of taxonomic changes recently approved by the ICTV. Archives of Virology 147(8), 1655–1663. https://doi.org/10.1007/s007050200039
MOHAMED, T.M., SUN, W., BUMBIE, G.Z., DOSOKY, W.M., RAO, Z., HU, P., WU, L., and TANG, Z., 2022. Effect of Dietary Supplementation of Bacillus subtilis on Growth Performance, Organ Weight, Digestive Enzyme Activities, and Serum Biochemical Indices in Broiler. Animals, 12(12), 1558. https://doi.org/10.3390/ani12121558
MOHAMED, T.M., SUN, W., BUMBIE, G.Z., ELOKIL, A.A., MOHAMMED, K.A., ZEBIN, R., HU, P., WU, L., and TANG, Z., 2022. Feeding Bacillus subtilis ATCC19659 to Broiler Chickens Enhances Growth Performance and Immune Function by Modulating Intestinal Morphology and Cecum Microbiota. Frontiers in Microbiology 12, 798350. https://doi.org/10.3389/fmicb.2021.798350
MOUNTZOURIS, K.C., TSITRSIKOS, P., PALAMIDI, I., ARVANITI, A., MOHNL, M., SCHATZMAYR, G., and FEGEROS, K., 2010. Effects of probiotic inclusion levels in broiler nutrition on growth performance, nutrient digestibility, plasma immunoglobulins, and cecal microflora composition. Poultry Science 89(1), 58–67. https://doi.org/10.3382/ps.2009-00308
NEETESON, A., AVENDAÑO, S., KOERHUIS, A., DUGGAN, B., SOUZA, E., MASON, J., RALPH, J., ROHLF, P., BURNSIDE, T., KRANIS, A., and BAILEY, R., 2023. Evolutions in Commercial Meat Poultry Breeding. Animals, 13(19), 3150. https://doi.org/10.3390/ani13193150
NRC., 1994. Nutrient Requirements of Poultry: National Research Council. Washington: National Academy Press.
OBERLÄNDER, B., FAILING, K., JÜNGST, C.M., NEUHAUS, N., LIERZ, M., and PALAU-RIBES, F.M., 2020. Evaluation of Newcastle Disease antibody titers in backyard poultry in Germany with a vaccination interval of twelve weeks. PLOS ONE 15(8), e0238068. https://doi.org/10.1371/journal.pone.0238068
OBERLÄNDER, J.C., KALTHOFF, D., and BEER, M., 2020. Newcastle disease virus: Systematic pathotyping and molecular characterization of avian isolates in Germany from 1939 to 2019. Viruses 12(7), 740. https://doi.org/10.3390/v12070740
OLADOKUN, S., and ADEWOLE, D., 2023. The effect of Bacillus subtilis and its delivery route on hatch and growth performance, blood biochemistry, immune status, gut morphology, and microbiota of broiler chickens. Poultry Science 102(4), 102473. https://doi.org/10.1016/j.psj.2022.102473
PARK J.H., and KIM I.H., 2014. Supplemental effect of probiotic Bacillus subtilis B2A on productivity, organ weight, intestinal Salmonella microflora, and breast meat quality of growing broiler chicks. Poultry Science 93, 2054–2059. https://doi.org/10.3382/ps.2013-03818
QIU, K., WANG, J., and GAO, J., 2021. Effects of dietary supplementation with Bacillus subtilis, as an alternative to antibiotics, on growth performance, serum immunity, and intestinal health in broiler chickens. Frontiers in Nutrition 8, 786878. https://doi.org/10.3389/fnut.2021.786878
RAMLUCKEN, U., LALLOO, R., ROETS, Y., MOONSAMY, G., VAN RENSBURG, C. J., and THANTSHA, M., 2020. Advantages of Bacillus-based probiotics in poultry production. Livestock Science 241, 104215. https://doi.org/10.1016/j.livsci.2020.104215
RATCLIFFE, M. J. 2005. Antibodies, immunoglobulin genes and the bursa of Fabricius in chicken B cell development. Developmental and Comparative Immunology 30(1–2), 101–118. https://doi.org/10.1016/j.dci.2005.06.018
REIS M.P., FASSANI E.J., JÚNIOR A.G., RODRIGUES P.B., BERTECHINI A.G., BARRETT N., BARRETT M.E., and SCHMIDT C.J., 2017. Effect of Bacillus subtilis (DSM 17299) on performance, digestibility, intestine morphology, and pH in broiler chickens. Journal of Applied Poultry Research 26, 573–583. https://doi.org/10.3382/japr/pfx032
RHAYAT, L., MARESCA, M., NICOLETTI, C., PERRIER, J., BRINCH, K. S., CHRISTIAN, S., DEVILLARD, E., and ECKHARDT, E., 2019. Effect of Bacillus subtilis strains on intestinal barrier function and inflammatory response. Frontiers in Immunology 10, 1–10. https://doi.org/10.3389/fimmu.2019.00564
ROSS, C.S., MAHMOOD, S., SKINNER, P., MAYERS, J., REID, S.M., ROWENA, D.E., HANSEN, R.D., and BANYARD, A.C., 2022. MM Profile: Avian paramyxovirus type-1 and Newcastle disease: a highly infectious vaccine-preventable viral disease of poultry with low zoonotic potential. Journal of Medical Microbiology 71, 001489. https://doi.org/10.1099/jmm.0.001489
SANDVANG, D., SKJOET-RASMUSSEN, L., CANTOR, M.D., MATHIS, G.F., LUMPKINS, B.S., and BLANCH, A., 2021. Effects of feed supplementation with 3 different probiotic Bacillus strains and their combination on the performance of broiler chickens challenged with Clostridium perfringens. Poultry Science 100(4), 100982. https://doi.org/10.1016/j.psj.2021.01.005
SHIM, Y.H., SHINDE, P.L., CHOI, J.Y., KIM, J.S., SEO, D.K., PAK, J.I., CHAE, B. J., and KWON, I.K., 2010. Evaluation of multimicrobial probiotics produced by submerged liquid and solid substrate fermentation methods in broilers. Asian-Australasian Journal of Animal Sciences, 23: 521–529. http://dx.doi.org/10.5713/ajas.2010.90446
SIKANDAR, A., ZANEB, H., NASIR, A., UR REHMAN, A., KASHIF. M., SHAH, M., LUQMAN, Z., DIN, S., IQBAL, M.F., KHAN, I., and IRSHAD, I., 2022. Effect of Bacillus subtilis on the microarchitectural development of the immune system in Salmonella-challenged broiler chickens. Veterinární Medicína-Czech67(1), 28–37. https://doi.org/10.17221/231/2020-VETMED
SPSS INC., 2015. SPSS FOR WINDOWS, VERSION 23.0. CHICAGO, IL: SPSS INC.
SUGIHARTO, S., YUDIARTI, T., ISROLI, I., WIDIASTUTI, E., and WAHYUNI, H. I., 2018. Hematological parameters and selected intestinal microbiota populations in the Indonesian indigenous crossbred chickens fed basal diet supplemented with multi-strain probiotic preparation in combination with vitamins and minerals. Veterinary World 11(6), 874. https://doi.org/10.14202/vetworld.2018.874-882
TIMMERMAN, H. M., KONINGB, C. J. M., MULDERC, L., ROMBOUTSD, F. M., and BEYNEN, A. C., 2004. Monostrain, multistrain and multispecies probiotics: A comparison of functionality and efficacy. International Journal of Food Microbiology 96, 219 ––233.
WANG, J., DENG, L., CHEN, M., CHE, Y., LI, L., ZHU, L., CHEN, G., and FENG, T., 2024. Phytogenic feed additives as natural antibiotic alternatives in animal health and production: A review of the literature of the last decade. Animal Nutrition, 17, 244-264. https://doi.org/10.1016/j.aninu.2024.01.012
WANG, Y., GU, Q., and LI, X., 2018. Use of probiotics in broiler chicken production: A review of the literature. Frontiers in Veterinary Science 5, 178. https://doi.org/10.3389/fvets.2018.00178
ZHANG, Z.F., CHO, J.H., and KIM, I.H., 2013. Effects of Bacillus subtilis UBT-MO2 on growth performance, relative immune organ weight, gas concentration in excreta, and intestinal microbial shedding in broiler chickens. Livestock Sci., 155: 343–347.