Effects of Bacillus subtilis on the immune parameters, intestinal morphology and mucin gene expression in broilers exposed to Salmonella enterica challenge
Published:
Jan 11, 2024
Keywords:
antibody titers bacterial population Immune response Intestinal morphology Probiotics
Abstract
The objective of this study was to evaluate the effect of the probiotic Bacillus subtilis supplementation on bacterial population, morphometry and mucin gene expression of intestine, and on immune response in Salmonella challenged broilers. Treatments were: 1) negative control (no probiotic- no challenging); 2) probiotic treated (no challenging); 3) positive control (no probiotic-salmonella challenging), and 4) Salmonella challenged chicks fed probiotic. Salmonella infection resulted in significant decrease in the relative weight of thymus. In challenged birds, dietary addition of probiotic increased (P<0.0.5) the relative weights of bursa and thymus. Population of lactic acid bacteria was significantly higher in probiotic treated groups as compared to negative control or challenged chickens. Challenged chickens had the lowest count of goblet cells and those received probiotic had the highest goblet cells count. The expression of mucin 2 gene was higher in group received dietary probiotic as compared with other treatments. Difference for gene expression was not found in challenged chickens and the negative control (P>0.05). In conclusion, the dietary inclusion of Bacillus subtilis could alleviate the negative effects of Salmonella infection on intestinal cells and bacterial population and could improve the growth and development of immune organs and function in infected broilers.
Article Details
- How to Cite
-
Jouybari, M., Sadeghi, A., Jouzani, G., Chamani, M., & Aminafshar, M. (2024). Effects of Bacillus subtilis on the immune parameters, intestinal morphology and mucin gene expression in broilers exposed to Salmonella enterica challenge . Journal of the Hellenic Veterinary Medical Society, 74(4), 6399–6410. https://doi.org/10.12681/jhvms.30558
- Issue
- Vol. 74 No. 4 (2023)
- Section
- Research Articles
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with this journal agree to the following terms:
· Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution Non-Commercial License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
· Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g. post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
· Authors are permitted and encouraged to post their work online (preferably in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.
Downloads
Download data is not yet available.
References
Abd El-Hack ME, El-Saadony MT, Shafi ME, Alshahrani OA,
SaghirSAM, Al-wajeeh AS, Al-shargi OYA, Taha AE, Mesalam
NM, Abdel-Moneim AE (2021) Prebiotics can restrict Salmonella populations in poultry: a review, Anim Biotechnol. doi:
1080/10495398.2021.1883637
Abudabos AM, Ali MH, Nassan MA, Saleh AA (2019) Ameliorative effect of Bacillus subtilis on growth performance and intestinal architecture in broiler infected with Salmonella. Animals. 9(4): 190. doi:
3390/ani9040190
Abudabos AM, Abdullah H, Alyemni Y, Dafalla M, Khan RU (2017)
Effect of organic acid blend and Bacillus subtilis alone or in combination on growth traits, blood biochemical and antioxidant status in broilers exposed to Salmonella typhimurium challenge
during the starter phase. J Applied Anim Res 45 (1): 538-542. doi:
1080/09712119.2016.1219665
Alkhalf A, Alhaj M, Al-Homidan I (2010). Influence of probiotic supplementation on immune response of broiler chicks. Egyptian Poult. Sci.
(1): 271-280
Awad WA, Aschenbach JR, Ghareeb K, Khayal B, Hess C, Hess M (2014)
Campylobacter jejuni influences the expression of nutrient transporter genes in the intestine of chickens. Vet Microbiol 172: 195-201. doi:
1016/j.vetmic.2014.04.001
Baltzley T, Lago F, Neumann T, Rehberger T, Gebert S (2010) Microorganisms and methods for treating poultry. Assignee Agtech Products
Inc. US Patent 7, 754,469.
Beachey EH (1981) Bacterial adherence: adhesin-receptor interactions
mediating the attachment of bacteria to mucosal surfaces. J Infect
Dis. 143:325-345.
CDC (2022) The book of 61st Conference on Decision and Control
(CDC), December 6-9, 2022, Cancun, Mexico.
Ciurescu G, Dumitru M, Gheorghe A, Untea AE, Drăghici R (2020) Effect of Bacillus subtilis on growth performance, bone mineralization,
and bacterial population of broilers fed with different protein sources.
Poult Sci 99(11): 5960-5971. doi: 10.1016/j.psj.2020.08.075
Close B, Banister K, Baumans V, Bernoth EM, Bromage N, Bunyan J,
(1997). Recommendations for euthanasia of experimental animals:
Part 2. Lab Anim. 31: 1-32.
Corrier DE, DeLoach JR (1990) Evaluation of cell-mediated, cutaneous
basophil hypersensivity in young chickens by aninterdigital skin test.
Poult Sci 69 (3): 403-408.
Dersjant-Li Y, Awati A, Kromm C, Evans C (2014) A direct fed microbial
containing a combination of three strain Bacillus sp. can be used as an
alternative to feed antibiotic growth promoter in broiler production. J
Appl Anim Nutr 2: 1-6.
Forder REA, Nattrass GS, Geier MS, .Hughes RJ, Hynd PI (2012) Quantitative analyses of genes associated with mucin synthesis of broiler
chickens with induced necrotic enteritis. Poult Sci 91 (6): 1335-1341.
Gaggia F, Mattarelli P, Biavati B (2010) Probiotics and prebiotics in animal feeding for safe food production. Inter J Food Microbiol 141:
S15-S28.
Gao Z, Wu H, Shi L, Zhang X, Sheng R, Yin F, Gooneratne R (2017)
Study of Bacillus subtilis on growth performance, nutrition metabolism and intestinal microflora of 1 to 42 d broiler hickens. Front.
Microbiol. 3(2): 109-113.
Grasman KA (2010) In vivo functional tests forassessing immunotoxicity
in birds. In: Dietert, RR (Ed.), Immunotoxicity testing: methods and
protocols, methods in molecular biology. (1st Edn.), New York, Humana Press. PP: 387-397.
Haghighi HR, Gong J, Gyles CL (2005) Modulation of antibody-mediated
immune response by probiotics in chickens. Clinic Diag Lab Immunol 12: 1387-1392.
Hans H. Stein & Dong Y. Kil. (2006) Reduced use of antibiotic growth
promoters in diets fed to weanling pigs: dietary tools. Anim Biotechnol. 17:2, 217-231, doi: 10.1080/10495390600957191.
Hayashi RM, Lourenço MC, Kraieski AL, Araujo RB, Esquerra RG,
Leonardecz E, da Cunha AF, Carazzolle MF, Monzani PS, Santin E
(2018) Effect of feeding Bacillus subtilis spores to broilers challenged
with Salmonella enterica serovar Heidelberg Brazilian Strain UFPR1
on performance, immune response, and gut health. Front Vet Sci 13:
doi: 10.3389/fvets.2018.00013
Hu XF, Guo YM, Huang BY, Zhang LB, Bun S, Liu D, Long FY, Li JH,
Yang X, Jiao P (2009) Effect of corticosterone administration on small
intestinal weight and expression of small intestinal nutrient transporter mRNA of broiler chickens. Asian-Aust J Anim Sci. 23(2):175-81.
Iji PA, Saki AA, Tivey DR (2001) Intestinal development and body
growth of broiler chicks on diets supplemented with non-starch
polysaccharides. Anim Feed Sci Technol 89: 175-188. doi: 10.1016/
s0377-8401(00)00223-6.
Khaksefidi A, Ghoorchi T (2006) Effect of probiotic on performance and
immunocompetence in broiler chicks. J Poult Sci 43: 296-300.
Kim DH, Austin B (2006) Cytokine expression in leucocytes and gut cells
of rainbow trout, Oncorhynchus mykiss Walbaum, induced by probiotics. Vet Immunol Immunopathol 114: 297-304.
Kim D, Min Y, Suminda GGD, Hur CG, Lee SC, Lee HK, Song KD, Heo J,
Son YO, Lee DS (2022) Bacillus-supplemented diet improves growth
performance in Jeju native pigs by modulating myogenesis and adipogenesis, Anim Biotechnol. doi: 10.1080/10495398.2022.2047996
Knap I, Kehlet AB, Bennedsen M, Mathis GF, Hofacre CL, Lumpkins BS
(2011) Bacillus subtilis (DSM17299) significantly reduces Salmonella in broilers. Poult Sci. 90: 1690-1694. doi:10.3382/ps.2010-01056.
Knarreborg A, Brockmann E, Høybye K, Knap I, Lund B, Milora N
(2008) Bacillus subtilis (DSM17299) modulates the ileal microbial
communities and improves growth performance in broilers. Int J Prebiotic Probiotic 3: 83-88.
Knarreborg A, Brockmann E, Høybye K, Knap I, Lund B, Milora N, Leser
TD (2008) Bacillus subtilis (DSM17299) modulates the ileal microbial communities and improves growth performance in broilers. Int J
Prebiotic. 3: 83-88
La Ragione RM, Woodward MJ (2003) Competitive exclusion by Bacillus
subtilis spores of Salmonella enterica serotype Enteritidis and Clostridium perfringens in young chickens. Vet Microbial 94: 245-256.
doi:10.1016/S0378-1135(03)00077-4.
Lipiński K, Mazur-Kuśnirek M, Antoszkiewicz Z, Makowski Z, Śliżewska K, Siwicki A, Otrocka-Domagała I, Gesek M (2021) The effect of
synbiotics and probiotics on the growth performance, gastrointestinal
function and health status of turkeys. Arch Anim Nut 75(5): 376-388.
doi: 10.1080/1745039X.2021.1958646
Lourenco MC, Kuritza LN, Westphal P, Muniz E, Pickler L, Santin E
(2012) Effects of Bacillus subtilis in the dynamics of infiltration of
immunological cells in the intestinal mucosa of chickens challenged
with Salmonella minnesota. Inernational J Poult Sci 11 (10): 630-634.
Ma Y, Wang W, Zhang H (2018) Supplemental Bacillus subtilis DSM
manipulates intestinal structure and microbial composition in
broiler chickens. Sci Rep 8:15358. doi: 10.1038/s41598-018-33762-8
Mack DR, Ahrne S, Hyde L, Wei S, Hollingsworth MA (2003) Extracellular MUC3 mucin secretion follows adherence of Lactobaccilus strains
to intestinal epithelial cells in vitro. Gut 52: 827-833.
Majidi-Mosleh A, Sadeghi AA, Mousavi SN, Chamani M, Zarei A.
(2016) Ileal MUC2 gene expression and microbial population, but
not growth performance and immune response influenced by in ovo
injection of probiotics in broiler chicks. Brit Pout Sci 58:1, 40-45,
doi: 10.1080/00071668.2016.1237766.
Nair AV, Leo Antony M, Praveen NK, Sayooj P, Raja Swaminathan T,
Vijayan KK (2021) Evaluation of in vitro and in vivo potential of
Bacillus subtilis MBTDCMFRI Ba37 as a candidate probiont in fish
health management. Microb Pathog. 152:104610. doi: 10.1016/j.micpath.2020.104610.
Oh J, Pajarillo EAB, Chae J (2017) Effects of Bacillus subtilis CSL2 on
the composition and functional diversity of the faecal microbiota of
broiler chickens challenged with Salmonella gallinarum. J Anim Sci
Biotechnol 8: 1. doi: 10.1186/s40104-016-0130-8
Post J, Rebel JMJ, Huurne AAHM (2003) Physiological effects of elevated plasma corticosterone concentrations in broiler chickens. An alternative means by which to assess the physiological effects of stress.
Poult Sci 82: 1313-1318.
Rahimi S, Khaksefidi A (2010) A comparison between the effects of a
probiotic (Bioplus 2B) and an antibiotic (virginiamycin) on the performance of broiler chickens under heat stress condition. Iranian J
Vet Res 7: 23-28.
Rahimi S, Kathariou S, Grimes JL, Siletzky RM (2011) Effect of direct-fed microbials on performance and Clostridium perfringens colonization of turkey poults. Poult Sci 90, 2656-2662. doi: 10.3382/
ps.2011-01342.
Ramakers C, Ruijter JM, Deprez RH, Moorman AF (2003) Assumption-free analysis of quantitative real-time polymerase chain reaction
(PCR) data. Neurosci Lett 339: 62-66.
Reid G, Friendship R (2002) Alternatives to antibiotic use: probiotics
for the gut. Anim Biotechnol 13:1, 97-112. doi: 10.1081/ABIO120005773
Riddell C (1987) Avian Histopathology. American Association of Avian
Pathologists, Kennett Square, PA. USA. PP: 10.
Saleem G, Ramzaan R, Khattak F, Akhtar R (2016) Effects of acetic acid
supplementation in broiler chickens orally challenged with Salmonella pullorum. Turk J Vet Anim Sci 40, 434-43. doi: 10.3906/vet1505-
SAS Institute. (2010). SAS®/STAT Software, Release 9.4. SAS Institute,
Inc., Cary, NC. USA
Schierack P, Wieler LH, Taras D (2007) Bacillus cereus var. toyoi enhanced systemic immune response in piglets. Vet Immunol Immunopathol 118: 1-11
Seifert S, Fritz C, Carlini N (2011) Modulation of innate and adaptive immunity by the probiotic Bifidobacterium longum PCB133 in turkeys.
Poult Sci 90: 2275-2280.
Shabani R, Fakhraei J, Yarahmadi HM, Seidavi A (2019) Effect of different sources of selenium on performance and characteristics of immune system of broiler chickens. Rev Brasil Zootec 48. 23-44.
Sharma R, Schumacher U (1995) Morphometric analysis of intestinal mucins under different dietary conditions and gut flora in rats. Dig Dis
Sci 40: 2532-2539.
Sikandar A, Zaneb H, Younus M, Masood S, Aslam A, Shah M, Rehman
H (2017) Growth performance, immune status and organ morphometry in broilers fed Bacillus subtilis-supplemented diet. S Afr J Anim
Sci 47(3), 378-88.
Sikandar A, Zaneb H, Nasir A, Rehman A, Kashif M, Shah M, Luqman
Z, Din S, Iqbal MF, Khan I, Irshad I (2020) Effect of Bacillus subtilis
on the microarchitectural development of the immune system in Salmonella-challenged broiler chickens. Vet Med-Czech 67: 28-37. doi:
17221/231/2020-VETMED.
Sikandar A, Zaneb H, Nasir A, Adil M, Ali HM, Muhammad M, Rehman
T, Rehman A, Rehman HF (2020) Effects of Bacillus subtilis on performance, immune system and gut in Salmonella-challenged broilers.
S Afr JA anim Sci 50(5). doi: 10.4314/sajas.v50i5.2
Swaggerty CL, Kaiser P, Rothwell L (2006) Heterophil cytokine mRNA
profiles from genetically distinct lines of chickens with differential
heterophil-mediated innate immune responses. Avian Pathol 35: 102-
Tarradas J, Tous N, Esteve-Garcia E (2020) The control of intestinal inflammation: A major objective in the research of probiotic strains as
alternatives to antibiotic growth promoters in poultry. Microorganisms 8(2): 148. doi: 10.3390/microorganisms8020148
Tsirtsikos P, Fegeros K, Balaskas C, Kominakis A, Mountzouris KC
(2012) Dietary probiotic inclusion level modulates intestinal mucin
composition and mucosal morphology in broilers. Poultry Science,
: 1860-1868.
Van Immerseel F, Russell JB, Flythe MD, Gantois I, Timbermont L, Pasmans F, Haesebrouck F, Ducatelle R (2006) The use of organic acids
to combat Salmonella in poultry: A mechanistic explanation of the efficacy. Avian Pathol 35: 182-188. doi: 10.1080/03079450600711045
Vicente JL, Torres-Rodriguez A, Higgins SE, Pixley C, Tellez G, Annie
Donoghue M, Hargis BM (2008) Effect of a selected Lactobacillus
spp.: based probiotic on Salmonella enterica Serovar Enteritidis: Infected broiler chicks .Avian Diseases 52 (1): 143-146.
Wibisono FM, Wibisono FJ, Effendi MH, Plumeriastuti H, Hidayatullah
AR, Hartadi EB, Sofiana ED (2020) A review of salmonellosis on
poultry farms: Public health importance. Syst Rev Pharm.;11: 481-
Witkamp M, Olson JS (1963) Breakdown of confined and non-confined
oak litter. Oikos 14: 138-147.
Yang Z, Liao SF (2019) Physiological effects of dietary amino acids on
gut health and functions of swine. Frontiers Veterinary Sci 2019; 6.
doi: 10.3389/fvets.2019.00169
Yang Y, Ruiping S, ShiMin Z, Yibao J (2007) Effect of probiotics on intestinal mucosal immunity and ultrastructure of cecal tonsils of chickens.
Poult Sci 121: 237-246, https://doi.org/10.1080/17450390500216928
Zhang L, Zhang L, Zeng X, Zhou L, Cao G, Yang C (2016) Effects of
dietary supplementation of probiotic, Clostridium butyricum, on
growth performance, immune response, intestinal barrier function,
and digestive enzyme activity inbroiler chickens challenged with
Escherichia coli K88. J. Anim. Sci. Biotechnol. 7(3): 1-9.
Zhenhua G, Wu H, Xiaohui S, RanShenga Z, YinaRavi F (2017) Study of
Bacillus subtilis on growth performance, nutrition metabolism and
intestinal microflora of 1 to 42 d broiler chickens. Anim Nut 3(2):
-113. doi: 10.1016/j.aninu.2017.02.002
Zhou A, Zhang X, Zhou Y, Xiao L, Li T (2021) Effect of direct-fed
microbials on growth performance, blood biochemical indices, and immune status of female goats, Anim Biotechnol. doi:
1080/10495398.2021.2014856
