Effects of in-ovo injection of silver nanoparticles on immune system function in broiler chickens under oxidative stress induced by lipopolysaccharide

Document Type : Research Paper

Authors

1 Gorgan University of Agricultural Sciences and Natural Resources

2 gorgan university

3 عضو هیات علمی

4 Vet, Maneger of Golestan Branch of IMI Gorgan, Iran

Abstract

Introduction: Stress, as a response to adverse stimuli, is difficult to define and understand mostly due to its nebulous perception. As Selye (1976) pointed out, “stress is the nonspecific response of the body to meet any demand”, whereas stressor is defined as “an agent that produces stress at any time”. Hence, stress represents the reaction of the animal organism (i.e., a biological response) to stimuli that disturb its normal physiological equilibrium or homeostasis. Thanks to their antioxidants, nutrients and anti-inflammatory properties silver nanoparticles are used in many developed countries (Bhanja et al 2015). The list potential applications of nanotechnology are very wide and diverse, but it is undoubtedly one of its most valuable applications in the development of therapeutic and pharmaceutical cases. Also, the use of silver nanoparticles in the treatment of deadly diseases like avian influenza supports further research on biological systems (Xiang et al 2011). Among inorganic materials, metal oxides such as TiO2, ZnO, MgO CaO, and Ag are of particular interest because they are stable in hard conditions and generally considered to be safe substances for humans and animals.
Material and methods: A total number of 592 eggs with an average weight of 50 g were purchased from a commercial Hubbard F15 broiler breeder flock aged 50 wk. The eggs, allotted to 4 treatments of 4 replicates with 37 eggs each, were set on the same floor to provide similar incubation conditions. Treatments including 2 doses of silver nanoparticles (20 and 40 mg) was injected via the egg holes using 1 mL insulin syringes equipped with disposable needles (21 gauge). The control group did not receive any treatment, and the sham control was injected with 1 ml phosphate-buffered saline (Triplett et al 2018). The injection holes were immediately covered using paraffin. At the end of d 18 of incubation, the eggs were transferred to the hatching cabinet. On the 21d of incubation, the hatched chicks were taken out of the incubator and after counting (checking for hatching) and weighting, half of the chicks were killed on the same day to check Hemoral immunity (G and M),white blood cell count and study of the desired genes (TNF-α, IL-6 and TGF-β). Among other chickens, those that were more uniform in weight with the desired treatment were transferred to the breeding period for 42 day and again in 4 treatments (treatments applied during incubation) and 4 replications with 20 broilers per replication. During 42 day, broilers were provided with unlimited water and food. To induce oxidative stress during the breeding period, 500 μg/kg live weight Salmonella lipopolysaccharide injected at 12, 24 and 48 hours before killed (Xi et al 2000).
Result and discussion: While ovo injection, silver nanoparticles was significantly decreased hatchability in comparison to the control group (P<0/05). In line with our results (Goel et al 2017) found that the hatchability of the eggs injected with nanoparticles was significantly lower than the control group. The reason can be attributed to the smaller size of younger embryos at the 7th ED, injection method, location injection, injection depth, injection time, genetics, hen age, egg size and hatching conditions (Sun et al 2018; Pilarski et al 2005). In case of ovo injection of 20 and 40 mg silver nanoparticles, significantly increased carcass percentage compared to the control group at hatching and post-hatch period, respectively (P<0/05). (Bhanja et al 2015; Pineda et al 2012) proved that silver nanoparticles had positive effects on embryo weight. Due to the significant amount of antioxidant in the silver nanoparticles inside the egg and its antioxidant effect on energy efficiency during embryonic life, silver nanoparticles receiving groups had a higher body weight at hatching time. Higher birth weight makes it possible to increase feed intake and weight gain in these treatments.
Upon ovo administration of 20 mg silver nanoparticles, spleen was increased significantly compared to the control group at hatching period (P<0/05). This is in line with a previous study of Goal et al (2017), who observed higher liver and spleen weights in 40 mg/kg silver nanoparticles group. The development of B and T lymphocytes initiates during embryogenesis in the bursa of Fabricius and thymus, respectively, and matures in the spleen until post-hatch (Erf 1997). These organs play an important role in imparting immunity. The cells produced in these organs differentiate into cellular immunity and humoral immunity, thus imparting immunity against different pathogens. Therefore, increased liver and spleen weight indicates a better immunological health status of in ovo silver nanoparticles supplemented birds. Ovo treatments did not affect the concentrations of immunoglobulin (IgG), (IgM), WBC counts and H/L ratio (P>0/05). Our results are consistent with previous studies of Pineda et al (2012) who showed the concentration of IgG and IgM were not affected by ovo injection of 10 and 20mg/kg silver nanoparticles. (Salari et al 2016; Saki and Salari 2015) showed silver nanoparticles increased in serum IgM and IgE, and increased in blood neutrophilic granulocytes. Rezaei Zarchi et al (2012) reported, feeding the rats for 28 days at doses of 25, 50, 100 and 200 mg / kg of silver nanoparticles had no significant effect on WBC and H/L. Differences in results may be due to differences in injection method, location injection, injection depth, injection time, genetics, hen age, egg size and hatching conditions. In comparison with the control and 40 mg silver nanoparticles group at hatching, there was significant up-regulation of TNF-α, IL-6 and TGF-β gene expression in 20 mg silver nanoparticles injected embryos,. Tumor necrosis factor-α (TNF-α) is a key cytokine involved in inflammation and immunity and the pro-inflammatory cytokine IL-6 induces the final maturation of B cells into antibody-secreting plasma cells, thereby increasing the secretion of immunoglobulins (Balkwill 2009; Mosmann 1989). Silver nanoparticles can interact with the immune system by binding and reacting with cells or proteins, thereby modulating the immune response. In the present study, higher expression of TNF-α gene in the livers was observed in ovo injected silver nanoparticles embryos. These findings also support earlier studies by Khan et al (2013), who showed the gene expression of IL-6 and TNF-𝛼 were affected by 50nm GNPs in the kidneys of rats. Also Vadalasetty et al (2018) reported that expression of TNF-α and NF-kB at mRNA were significantly up-regulated in the 50ppm silver nanoparticles group. Conclusion: The results of this study suggest that silver nanoparticles improve the immune response of broilers by improving growth and increasing the relative expression of genes involved in immune function.

Keywords


Ahmadi J, 2009. Aplication of different levels of silver nanopracticles in food on the performance and some blood parameters of broiler chickens. World Applied Science Journal 7: 24-27.
Alqazlan N, Alizadeh MA, Boodhoo N, Taha-Abdelaziz KH, Nagy E, Bridle B and Sharif  SH, 2020. Probiotic Lactobacilli Limit Avian Influenza Virus Subtype H9N2 Replication in Chicken Cecal Tonsil Mononuclear Cells. Vaccines 8:2-14.
Andi MA, Mohsen H and Farhad A, 2011. Effects of feed type with /without nanosil on cumulative performance, relative organ weight and some blood parameters of broilers. Global Veterinaria 7: 605-609.
Bhanja SK, Anna H, Mehra M, Sawosz E, Pineda L, Vadalasetty KP,  Kurantowicz N and Chwalibog A, 2015. In Ovo Administration of Silver Nanoparticles and/or Amino Acids Influence Metabolism and Immune Gene Expression in Chicken Embryos. International Journal of Molecular Sciences 16: 9484-9503.
 De Jong WH, Van Der Ven LM, Sleijffers Park MVDZ, Jansen EHJM, Loveren HV and Vandebriel  RJ, 2013. Systemic and immunotoxicity of silver nanoparticles in an intravenous 28 days repeated dose toxicity study in ratsq. Biomaterials12:1-11.
Erf GF, 1997. Immune system function and development in broilers. Poultry Science 8:108-115.
Goel A, Bhanja SK, Mehra M, Majumdar S and Mandal A, 2017. In ovo silver nanoparticle supplementation for improving the post-hatch immunity status of broiler chickens. Journal of Animal Nutrition 71: 384–394.
 Granado M, Martin AI, Lopez Menduina M, Lopez Calderon A and Villanua MA, 2008. GH-releasing peptide-2 administration prevents liver inflammatory response in endotoxemia. American Journal of Physiology-Endocrinology and Metabolism 294(1): 131-41.
Hateifi A, Zare shahne A, Ansari pirsaraie Z, Alizadeh AM, Atashnak MP, Masoudi R and Pio F, 2021. The Combined Anti-inammatory Strategy of Beta-2 Adrenergic Agonist and Glucocorticoid on the Laying Hen Model of Ovarian Cancer: the Immune Traits and Ovarian In ammatory Functions. Research Square :231-19.
Iseri SO, Sener G, Saglam B, Ercan F, Gedik N and Yege BC, 2008. Ghrelin alleviates biliary obstruction-induced chronic hepatic injury in rats. Regulatory Peptides 146(1-3):73-9.
Jones SA, 2005. Directing transition from innate to acquired immunity, defining a role for IL-6. Immunology175: 3463–3468.
Khan AH, Abdelhalim MAK, Alhomida AS and Al-Ayed MS, 2013. Effects of Naked Gold Nanoparticles on Proinflammatory Cytokines mRNA Expression in Rat Liver and Kidney. BioMedical Research16: 1-6.
Kim S and Ryu DY, 2013.  Silver nanoparticle-induced oxidative stress, genotoxicity and apoptosis in cultured cells and animal tissues. Journal of Applied Toxicology 33(2):78–89. 10.1002/jat.2792.
Klippstein R, Fernandez-Montesinos R, Castillo PM, Zaderenko AP and Pozo D, 2010. Silver nanoparticles interactions with the immune system, implications for health and disease. In Silver Nanoparticles; Perez, D.P., Ed.; InTech: Croatia, Europe 16: 309–324.
Livak KJ and  Schmittgen TD, 2001. Analysis of Relative Gene Expression Data Using Real-
      Time Quantitative PCR and the 22DDCT Method. Methods 25: 402–408.
Pilarski R, Bednarczyk M, Lisowski A, Rutkowski Z, Bernacki M and  Gulewicz k, 2005. Assessment of the effect of alpha-galactosides injected during embryogenesis on selected chicken traits. Folia Biologica 53: 13-20.
Pineda L, Chwalibog A, Sawosz E, Lauridsen C, Engberg R, Elnif J, Hotowy A, Sawosz F, Gao Y, Ali A and Sepehri Moghadam H, 2012. Effect of silver nanoparticles on growyh, performance, metabolism and microbial profile of broiler chickens. Archives of Animal Nurition 66: 416-429.
Pisoschi AM and Pop A, 2015. The role of antioxidants in the chemistry of oxidative stress: a review. European Journal of Medicinal Chemistry 97: 55-74.
Saki AA and Salari J, 2013. In ovo injection of nano silver, thyme and savory extracts on the 17th day of embryo and its effect on performance and blood parameters of broilers on days 14 and 21 of breeding. Journal of Animal Science Research and Construction 7:101.
Salari J, Saki AA, Abasinejad M and Manafi M, 2015. In ovo injection of nano silver, thyme and savory extracts to broiler breeders eggs and their effect on post- hatch immunological parameters. Iranian Animal Science 108:95-100.
SAS Institute Inc, 2003. SAS Procedure Guide. Version 9.2. Cary, NC: SAS Institute Inc.
Selye H, 1976. Forty years of stress research: principal remaining problems and misconceptions.  Canadian Medical Association Journal 115: 53–56.
Shaowei Chen RS, Ingram Michael J, Hostetler JJ, Pietron RW, Murray T and Gregory Schaaff JT, 1998. Gold Nanoelectrodes of Varied Size: Transition to Molecule-Like Charging. Science, 2098-21010
Sun X, Lu X, Liao L, Zhang X, Lin X and Ma Q, 2018. Effect of in ovo zinc injection on the embryonic development and epigenetics-related indices of zinc-deprived broiler breeder eggs. Biological Trace Element Research 185(2): 456-46.
Triplett MD, Zhai W and Peebles ED, 2018. Investigating commercial in ovo tech-nology as a strategy for introducing probiotic bacteria to broiler embryos. Poultry Science 97: 658-666.
Vadalasetty KP, Lauridsen CH, Margarete Engberg  R, Vadalasetty R, Kutwin M, Chwalibog M and Sawosz E, 2018. Influence of silver nanoparticles on growth and health of broiler chickens after infection with Campylobacter jejuni. BMC Veterinary Research 1-11.
Van Oosten M, Van de Bilt E, Van Berkel TJ and Kuiper J, 1998. New scavenger receptor-like receptors for the binding of lipopolysaccharide to liver endothelial and kupffer cells. Infect Immun 66(11):5107-5112.
Xiang  D, Chen Q, Pang L and  Zheng C, 2011. Inhibitory effects of silver nanoparticles on H1N1 influenza a virus in vitro. Journal of Virological Methods 178: 137–142.
Xie H, Rath  NC, Huff  GR, Huff WE  and Balog  JM, 2000. Effects of Salmonella typhimurium Lipopolysaccharide on Broiler Chickens. Poultry Science 79:33–40.