Effect of OPG-supplemented diet on immune and stress responses and attenuation of LPS-induced damage in the liver of carp (Cyprinus carpio haematopterus)





Effects of oxidized peptidoglycan (OPG) on immune and stress responses and lipopolysaccharide (LPS)-induced damage in the liver of carp were investigated in this study. Four hundred carps (Cyprinus carpio haematopterus) were fed with five experimental diets supplemented with 0, 100, 200, 400, and 800 mg kg-1 OPG for 28 days. Each group had four replicates and 20 fish per replication. LPS challenge (injection of 40 mg kg-1 saline or LPS) occurred at day 29. The supplementation with OPG linearly increased (p<0.05) plasma total protein, immunoglobulin M (IgM), complement 4 (C4), cortisol, and lactate on day 14. Dietary supplementation with OPG linearly increased (p<0.05) plasma and complement 3 (C3); quadratically improved (p<0.05) alkaline phosphatase (ALP) and lysozyme (LYS) activities; linearly increased hepatic superoxide dismutase (SOD) and catalase (CAT) activities; increased malondialdehyde (MDA) contents; and improved (p<0.05) hepatic anti-superoxide anion (ASA) and anti-hydroxy radical (AHR) contents on days 14 and 28. Dietary OPG significantly prevented the increase of interleukin 1β (IL-1β), interleukin 6 (IL-6), and tumor necrosis factor α (TNF-α) by inhibiting the excessive activation of TLR2-Myd88 signaling pathway; downregulating TLR2, Myd88, and NF-κB p65; and upregulating nuclear factor erythroid-2-related factor 2 (Nrf2) and Keap1 mRNA expression (p<0.05). Therefore, this study indicated that dietary OPG improves the plasma immune response, regulates the hepatic antioxidant status, and attenuates LPS-induced negative effects in the carp at the optimal dose of 400 mg kg-1.


Angeles, D.M.; Liu, Y.; Hartman, A.M.; Borisova, M.; Borges, A.S.; Kok, N.D.; Beilharz, K.; Veening, J.W.; Mayer, C. 2017. Pentapeptide-rich peptidoglycan at the Bacillus subtilis cell-division site. Molecular Microbiology, 104(2): 319-333. https://doi.org/10.1111/mmi.13629

Armenteros, M.; Heinonen, M.; Ollilainen, V.; Toldrá, F.; Estévez, M. 2009. Analysis of protein carbonyls in meat products by using the DNPH-method, fluorescence spectroscopy and liquid chromatographye-electrospray ionisatione-mass spectrometry (LC-ESI-MS). Meat Science, 83(1): 104-112. https://doi.org/10.1016/j.meatsci.2009.04.007

Barka, T.; Anderson, P. 1962. Histochemical methods for acid phosphatase using hexazonium pararosanalin as coupler. Journal of Histochemistry & Cytochemistry, 10: 741-753.

Beeby, M.; Gumbart, J.C.; Roux, B.; Jensen, G.J. 2013. Architecture and assembly of the Gram-positive cell wall. Molecular Microbiology, 88(4): 664-672. https://doi.org/10.1111/mmi.12203

Beutler, E. 1975. Red cell metabolism: a manual of biochemical methods. New York: Grune Strottan.

Chen, X.M.; Wang, Q.J.; Guo, Z.X.; Zhao, Y.L.; Luo, S.; Yu, T.; Zhang, D.M.; Wang, G.Q. 2020. Identification of the Nrf2 in the fathead minnow muscle cell line: role for a regulation in response to H2O2 induced the oxidative stress in fish cell. Fish Physiology and Biochemistry, 46(5): 1699-1711. https://doi.org/10.1007/s10695-020-00822-8

China Society of Fisheries. 2020. China fisheries statistical yearbook. Beijing: China Agriculture Press.

Duan, X.D.; Feng, L.; Jiang, W.D.; Wu, P.; Liu, Y.; Kuang, S.Y.; Tang, L.; Tang, W.N.; Zhang, Y.A.; Zhou, X.Q. 2019. Dietary soybean β-conglycinin suppresses growth performance and inconsistently triggers apoptosis in the intestine of juvenile grass carp (Ctenopharyngodon idella) in association with ROS-mediated MAPK signaling. Aquaculture Nutrition, 25(4): 770-782. https://doi.org/10.1111/anu.12895

Ellis, A. 1990. Techniques in fish immunology. Lysozyme Assays, 101-103.

Giri, S.S., Sen, S.S.; Chi, C.; Kim, H.J.; Yun, S.; Park, S.C. 2015a. Chlorophytum borivilianum polysaccharide fraction provokes the immune function and disease resistance of Labeo rohita against Aeromonas hydrophila. Journal of Immunology Research, 6;2015: 256510. https://doi.org/10.1155/2015/256510

Giri, S.S.; Sen, S.S.; Chi, C.; Kim, H.J.; Yun, S.; Park, S.C.; Sukumaran, V. 2015b. Effects of intracellular products of Bacillus subtilis VSG1 and Lactobacillus plantarum VSG3 on cytokine responses in the head kidney macrophages of Labeo rohita. Fish & Shellfish Immunology, 47(2): 954-961. https://doi.org/10.1016/j.fsi.2015.10.033

Hamann, L.; EL-Samalouti, V.; Ulmer, A.J.; Flad, H.D.; Rietschel, E.T. 1998. Components of gut bacteria as immunomodulators. International Journal of Food Microbiology, 41(2): 141-154. https://doi.org/10.1016/S0168-1605(98)00047-6

Hao, J.; Wu, F.; Tang, R.; Sun, Y.; Liu, D.; Zhang, Z. 2020. Preparation of 1,4-linked α-D-glucuronans from starch with 4-acetamide-TEMPO/NaClO2/NaClO system. International Journal of Biological Macromolecules, 151: 740-746. https://doi.org/10.1016/j.ijbiomac.2020.02.211

He, J.Y.; Wu, Z.; Pan, D.D.; Guo, Y.X.; Zen, X.Q. 2017. Effect of selenylation modification on antitumor activity of peptidoglycan from Lactobacillus acidophilus. Carbohydrate Polymers, 165: 344-350. https://doi.org/10.1016/j.carbpol.2017.02.031

Isogai, A.; Saito, T.; Fukuzumi, H. 2011. TEMPO-oxidized cellulose nanofibers. Nanoscale, 2011;3(1): 71-85. https://doi.org/10.1039/c0nr00583e

Jia, R.; Du, J.L.; Cao, L.P.; Li, Y.; Johnson, O.; Gu, Z.; Jeney, G.; Xu, P.; Yin, G. 2019. Antioxidative, inflammatory and immune responses in hydrogen peroxide-induced liver injury of tilapia (GIFT, Oreochromis niloticus). Fish & Shellfish Immunology, 84: 894-905. https://doi.org/10.1016/j.fsi.2018.10.084

Jiang, J.; Shi, D.; Zhou, X.Q.; Hu, Y.; Feng, L.; Liu, Y.; Jiang, W.D.; Zhao, Y. 2015. In vitro and in vivo protective effect of arginine against lipopolysaccharide induced inflammatory response in the intestine of juvenile Jian carp (Cyprinus carpio var. Jian). Fish & Shellfish Immunology, 42(2): 457-464. https://doi.org/10.1016/j.fsi.2014.11.030

Lee, S.H.; Ingale, S.L.; Kim, J.S.; Kim, K.H.; Lokhande, A.; Kim, E.K.; Kwon, I.K.; Kim, Y.H.; Cha, B.J. 2014. Effects of dietary supplementation with Bacillus subtilis LS1–2 fermentation biomass on growth performance, nutrient digestibility, cecal microbiota and intestinal morphology of weanling pig. Animal Feed Science and Technology, 188: 102-110. https://doi.org/10.1016/j.anifeedsci.2013.12.001

Li, W.H.; Wang, L.; He, H.Y.; Chen, J.; Yu, Y.R. 2016. Expression of neutrophil gelatinase-associated lipocalin in low osmolar contrast-induced nephropathy in rats and the effect of Nacetylcysteine. Experimental and Therapeutic Medicine, 12(5): 3175-3180. https://doi.org/10.3892/etm.2016.3779

Liu, X.M.; Xu, Z.; Chang, X.Q.; Fang, J.K.H.; Song, J.; He, J.F.; Tai, Z.G.; Zhu, Q.G.; Hu, M.H. 2021. Enhanced immunity and hemocytes proliferation by three immunostimulants in tri-spine horseshoe crab Tachypleus tridentatus. Fish & Shellfish Immunology, 115: 112-123. https://doi.org/10.1016/j.fsi.2021.06.001

Livak, K.J.; Schmittgen, T.D. 2001. Analysis of relative gene expression data using realtime quantitative PCR and the 2∆∆CT method. Methods, 25(4): 402-408. https://doi.org/10.1006/meth.2001.1262

Manhar, A.K.; Bashir, Y.; Saikia, D.; Nath, D.; Gupta, K.; Konwar, B.K.; Kumar, N.; Namsa, N.D.; Mandal, M. 2017. Corrigendum to “Cellulolytic potential of probiotic Bacillus subtilis AMS6 isolated from traditional fermented soybean (Churpi): An in-vitro study with regards to application as an animal feed additive”. Microbiological Research, 198: 56-57. https://doi.org/10.1016/j.micres.2017.02.005

Matsumoto, S.; Hara, T.; Nagaoka, M.; Mike, A.; Mitsuyama, K.; Sako, T.; Yamamoto, M.; Kado, S.; Takada, T. 2009. A component of polysaccharide peptidoglycan complex on Lactobacillus induced an improvement of murine model of inflammatory bowel disease and colitis-associated cancer. Immunology, 128(1 Part 2): e170-e180. https://doi.org/10.1111/j.1365-2567.2008.02942.x

Ming, J.H.; Ye, J.Y.; Zhang, Y.X.; Xu, Q.Y.; Yang, X.; Shao, X.P.; Qiang, J.; Xu, P. 2020. Optimal dietary curcumin improved growth performance, and modulated innate immunity, antioxidant capacity and related genes expression of NF-κB and Nrf2 signaling pathways in grass carp (Ctenopharyngodon idella) after infection with Aeromonas hydrophila. Fish & Shellfish Immunology, 97: 540-553. https://doi.org/10.1016/j.fsi.2019.12.074

Pandeirada, C.O.; Merkx, D.W.H.; Janssen, H.G.; Westphal, Y.; Schols, H.A. 2021. TEMPO/NaClO2/NaOCl oxidation of arabinoxylans. Carbohydrate Polymers, 259: 117781. https://doi.org/10.1016/j.carbpol.2021.117781

Pionnier, N.; Falco, A.; Miest, J.; Frost, P.; Irnazarow, I.; Shrive, A. Hoole, D. 2013. Dietary β-glucan stimulate complement and C-reactive protein acute phase responses in common carp (Cyprinus carpio) during an Aeromonas salmonicida infection. Fish & Shellfish Immunology, 34(3): 819-831. https://doi.org/10.1016/j.fsi.2012.12.017

Reuter, G. 2001. The Lactobacillus and Bifidobacterium microflora of the human intestine: Composition and succession. Current Issues in Intestinal Microbiology, 2(2): 43-53. https://doi.org/10.1002/1097-0142(195701/02)10:13.0.CO;2-V

Sheng, Q. 2011. Extraction of Peptidoglycan from Bacillus subtilis and its effects on immune function of mice. Sichuan Agricultural University.

Shi, H.Y.; Guo, Y.M.; Liu, Y.; Shi, B.L.; Guo, X.Y.; Jin, L.; Yan, S.M. 2016. The in vitro effect of lipopolysaccharide on proliferation, inflammatory factors and antioxidant enzyme activity in bovine mammary epithelial cells. Animal Nutrition, 2(2): 99-104. https://doi.org/10.1016/j.aninu.2016.03.005

Silvestre, F. 2020. Signaling pathways of oxidative stress in aquatic organisms exposed to xenobiotics. Journal of Experimental Zoology Part A: Ecological and Integrative Physiology, 333(6): 436-448. https://doi.org/10.1002/jez.2356

Takemura, A. 1993. Changes in an immunoglobulin M (IgM)-like protein during larval stages in tilapia, Oreochromis mossambicus. Aquaculture, 115(3-4): 233-241. https://doi.org/10.1016/0044-8486(93)90139-P

Tang, H.G.; Wu, T.X.; Zhao, Z.Y.; Pan, X.D. 2008. Effects of fish protein hydrolysate on growth performance and humoral immune response in large yellow croaker (Pseudosciaena crocea R.). Journal of Zhejiang University-Science B, 2008;9: 684-690. https://doi.org/10.1631/jzus.B0820088

Tiantian, Z.; Haicheng, Y.; Wei, H. 2019. Protective Effect of Bacillus subtilis Peptidoglycan (PG) on β-conglycinin-induced intestinal epithelial cells damage of juvenile carp (Cyprinus carpio). Animal Husbandry and Feed Science, 2019;11(2): 56-61. https://doi.org/10.19578/j.cnki.ahfs.2019.02.004

Wei, K.Q.; Yang, J.X.; Song, C.X. 2020. The responses of prophenoloxidase and MAPK/Nrf2 pathway to cadmium stress in red swamp crayfish Procambarus clarkii. Marine and Freshwater Behaviour and Physiology, 53(2): 59-72. https://doi.org/10.1080/10236244.2020.1764189

Wu, Z.; Pan, D.D.; Guo, Y.X.; Zeng, X.Q. 2013. Structure and anti-inflammatory capacity of peptidoglycan from Lactobacillus acidophilus in RAW-264.7 cells. Carbohydrate Polymers, 96(2): 466-473. https://doi.org/10.1016/j.carbpol.2013.04.028

Wu, Z.; Pan, D.D.; Zeng, X.Q.; Sun, Y.Y.; Cao, J.X. 2016. Phosphorylation of peptidoglycan from Lactobacillus acidophilus and its immunoregulatory function. International Journal of Food Science & Technology, 51(3): 664-671. https://doi.org/10.1111/ijfs.13028

Yu, Y.B.; Wang, C.H. Wang, A.M.; Yang, W.P.; Lv, F.; Liu, F.; Liu, B.; Sun, C.X. 2018. Effects of various feeding patterns of Bacillus coagulans on growth performance, antioxidant response and Nrf2-Keap1 signaling pathway in juvenile gibel carp (Carassius auratus gibelio). Fish & Shellfish Immunology, 73: 75-83. https://doi.org/10.1016/j.fsi.2017.11.050

Yu, Z.; Zheng, Y.G.; Du, H.L.; Li, H.J.; Wu, L.F. 2020. Bioflocs protects copper-induced inflammatory response and oxidative stress in Rhynchocypris lagowski Dybowski through inhibiting NF-κB and Nrf2 signaling pathways. Fish & Shellfish Immunology, 98: 466-476. https://doi.org/10.1016/j.fsi.2020.01.048

Zhang, X.D.; Zhu, Y.F.; Cai, L.S.; Wu, T.X. 2008. Effects of fasting on the meat quality and antioxidant defenses of market-size farmed large yellow croaker (Pseudosciaena crocea). Aquaculture, 280(1-4): 136-139. https://doi.org/10.1016/j.aquaculture.2008.05.010

Zhao, J.; Liu, Y.; Jiang, J.; Wu, P.; Jiang, W.D.; Li, S.H.; Tang, L.; Kuang, S.Y.; Feng, L.; Zhou, X.Q. 2013. Effects of dietary isoleucine on the immune response, antioxidant status and gene expression in the head kidney of juvenile Jian carp (Cyprinus carpio var. Jian). Fish & Shellfish Immunology, 35(2): 572-580. https://doi.org/10.1016/j.fsi.2013.05.027

Zhou, Z.X.; Lin, Z.J.; Pang, X.; Shan; P.P.; Wang, J.X. 2018. MicroRNA regulation of Toll-like receptor signaling pathways in teleost fish. Fish & Shellfish Immunology, 75: 32-40. https://doi.org/10.1016/j.fsi.2018.01.036






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