Skip to main content

Advertisement

SpringerLink
Log in

A study of the damage of the intestinal mucosa barrier structure and function of Ctenopharyngodon idella with Aeromonas hydrophila

  • Published:
Fish Physiology and Biochemistry Aims and scope Submit manuscript

Abstract

The aim of this study is to explore the effect of Aeromonas hydrophila on the intestinal mucosal barrier structure and intestinal permeability in grass carp (Ctenopharyngodon idella). Histopathological examinations showed that A. hydrophila induced severe intestinal lesions, including inflammatory cell infiltration and intestinal villus fusion and swelling. Messenger RNA (mRNA) expression of the inflammatory cytokines TNF-α, IL-1β, IL-8, IL-10 and MyD88 was significantly increased after infection with A. hydrophila. The permeability of intestinal mucosa was determined using Evans blue (EB) and d-lactic acid. The results indicated that the levels of EB and serum d-lactic acid were significantly increased after infection with A. hydrophila (p < 0.05). Our results also indicated that the intestinal mucosal barrier injury induced by A. hydrophila infection was closely associated with the expression of the tight junction (TJ) protein zonula occludens-1 (ZO-1), occludin, claudin b and claudin c as well as the activity of Na+, K+-ATPase and Ca2+, Mg2+-ATPase. Lower mRNA levels of occludin and lower Na+, K+-ATPase and Ca2+, Mg2+-ATPase activity in the intestines were observed after challenge. ZO-1 and claudin c were significantly increased 24 h after infection with A. hydrophila. The most interesting finding was that claudin b also significantly increased 24 h after challenge and then decreased to lower levels at 72, 120 and 168 h post-infection compared to the PBS-treated control group. The results demonstrated that grass carp infection with A. hydrophila induced intestinal inflammation and impaired the structure and function of the intestinal mucosal barrier.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Alsadi RM, Ma TY (2007) IL-1β causes an increase in intestinal epithelial tight junction permeability. J Immunol 178:4641–4649

    Article  CAS  Google Scholar 

  • Berkes J, Viswanathan VK, Savkovic SD, Hecht G (2003) Intestinal epithelial responses to enteric pathogens: effects on the tight junction barrier, ion transport, and inflammation. Gut 53:439–451

    Article  Google Scholar 

  • Bücker R, Krug SM, Rosenthal R, Günzel D, Fromm A, Zeitz M et al (2011) Aerolysin from Aeromonas hydrophila perturbs tight junction integrity and cell lesion repair in intestinal epithelial HT-29/B6 cells. J Infect Dis 204:1283–1292

    Article  PubMed  Google Scholar 

  • Capaldo CT, Nusrat A (2009) Cytokine regulation of tight junctions. Biochim Et Biophys Acta 1788:864–871

    Article  CAS  Google Scholar 

  • Cascn A, Yugueros J, Temprano A, Snchez M, Hernanz C, Luengo JM et al (2000) A major secreted elastase is essential for pathogenicity of Aeromonas hydrophila. Infect Immun 68:3233–3241

    Article  Google Scholar 

  • Chasiotis H, Kelly SP (2011) Permeability properties and occludin expression in a primary cultured model gill epithelium from the stenohaline freshwater goldfish. J Comp Physiol B 181:487–500

    CAS  PubMed  Google Scholar 

  • Chasiotis H, Kolosov D, Bui P, Kelly SP (2012) Tight junctions, tight junction proteins and paracellular permeability across the gill epithelium of fishes: a review ☆. Respir Physiol Neurobiol 184:269–281

    Article  CAS  PubMed  Google Scholar 

  • Chen L, Feng L, Jiang WD, Jiang J, Wu P, Zhao J et al (2015) Intestinal immune function, antioxidant status and tight junction proteins mRNA expression in young grass carp (Ctenopharyngodon idella) fed riboflavin deficient diet. Fish & Shellfish Immunology 47:470–484

    Article  CAS  Google Scholar 

  • Da Silva BC, Mourino JLP, Vieira FN, Jatoba A, Seiffert WQ, Martins ML (2012) Haemorrhagic septicaemia in the hybrid surubim (Pseudoplatystoma corruscans x Pseudoplatystoma fasciatum) caused by Aeromonas hydrophila. Aquacuture Research 43:908–919

    Article  Google Scholar 

  • Deitch EA (2002) Bacterial translocation or lymphatic drainage of toxic products from the gut: what is important in human beings? Surgery 131:241–244

    Article  PubMed  Google Scholar 

  • Ferguson RM, Merrifield DL, Harper GM, Rawling MD, Mustafa S, Picchietti S et al (2010) The effect of pediococcus acidilactici on the gut microbiota and immune status of on-growing red tilapia (Oreochromis niloticus). J Appl Microbiol 109:851–862

    Article  CAS  PubMed  Google Scholar 

  • Groschwitz KR, Hogan SP (2009) Intestinal barrier function: molecular regulation and disease pathogenesis. J Allergy Clin Immunol 124:3–22

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Huang RW, Ye YT, Cai CF, Wu P, Chen KQ, Wu T et al (2015) The study on damage of intestinal mucosa barrier structure with oxidized fish oil diets in Ctenopharyngodon idella. J Fish China 39:1511–1520

    Google Scholar 

  • Jr BR, Reynolds JJ, Bearden SE (2011) Hyperhomocysteinemia increases permeability of the blood-brain barrier by NMDA receptor-dependent regulation of adherens and tight junctions. Blood 118:2007–2014

    Article  Google Scholar 

  • Jutfelt F, Sundh H, Glette J, Mellander L, Björnsson BT, Sundell K (2008) The involvement of Aeromonas salmonicida, virulence factors in bacterial translocation across the rainbow trout, Oncorhynchus mykiss (walbaum) intestine. J Fish Dis 31:141–151

    Article  CAS  PubMed  Google Scholar 

  • Kolosov D, Bui P, Chasiotis H, Kelly SP (2013) Claudins in teleost fishes. Tissue Barriers 1:80–88

    Article  Google Scholar 

  • Konno H, Yamamoto T, Yamazaki K, Gohda J, Akiyama T, Semba K et al (2009) TRAF 6 establishes innate immune responses by activating NF-kappa B and IRF 7 upon sensing cytosolic viral RNA and DNA. PLoS One 4:e5674–e5674

    Article  PubMed  PubMed Central  Google Scholar 

  • Kumai Y, Bahubeshi A, Steele S, Perry SF (2011) Strategies for maintaining Na+, balance in zebrafish (Danio rerio) during prolonged exposure to acidic water. Comparative Biochemistry & Physiology Part A Molecular & Integrative Physiology 160:52–62

    Article  CAS  Google Scholar 

  • Kwong RW, Perry SF (2013) The tight junction protein claudin-b regulates epithelial permeability and sodium handling in larval zebrafish, Danio rerio. American Journal of Physiology Regulatory Integrative & Comparative Physiology 304:R504–R513

    Article  CAS  Google Scholar 

  • Lange S, Delbro DS, Jennische E (2009) Evans blue permeation of intestinal mucosa in the rat. Scand J Gastroenterol 29:38–46

    Article  Google Scholar 

  • Lee KK, Liu PC, Chuang WH (2002) Pathogenesis of gastroenteritis caused by Vibrio carchariae in cultured marine fish. Mar Biotechnol 4:267–277

    Article  CAS  PubMed  Google Scholar 

  • Li W, Zhou XQ, Feng L, Liu Y, Jiang J (2010) Effect of dietary riboflavin on growth, feed utilization, body composition and intestinal enzyme activities of juvenile jian carp (Cyprinus carpio, var. jian). Aquac Nutr 16:137–143

    Article  CAS  Google Scholar 

  • Lingrel JB (2010) The physiological significance of the cardiotonic steroid/ouabain-binding site of the Na+, k+-ATPase. Physiology 72:395–412

    Article  CAS  Google Scholar 

  • Liu HP, Hu CH, Xu Y (2008) Effects of early weaning on intestinal permeability and tight junction protein occludin mRNA expression levels of piglets. Chinese Journal of Animal Nutrition 20:442–446

    CAS  Google Scholar 

  • Liu X, Chang X, Wu H, Xiao J, Gao Y, Zhang Y (2014) Role of intestinal inflammation in predisposition of Edwardsiella tarda, infection in zebrafish (Danio rerio). Fish & Shellfish Immunology 41:271–278

    Article  Google Scholar 

  • Lü A, Hu X, Zheng L, Zhu A, Cao C, Jiang J (2011) Isolation and characterization of citrobacter spp. from the intestine of grass carp Ctenopharyngodon idellus. Aquaculture 313:156–160

    Article  Google Scholar 

  • Luo JB, Feng L, Jiang WD, Liu Y, Wu P, Jiang J et al (2014) The impaired intestinal mucosal immune system by valine deficiency for young grass carp (Ctenopharyngodon idella) is associated with decreasing immune status and regulating tight junction proteins transcript abundance in the intestine. Fish & Shellfish Immunology 40:197–207

    Article  CAS  Google Scholar 

  • Ma TY, Tran D, Hoa N, Nguyen D, Merryfield M, Tarnawski A (2000) Mechanism of extracellular calcium regulation of intestinal epithelial tight junction permeability: role of cytoskeletal involvement. Microscopy Research & Technique 51:156–168

    Article  CAS  Google Scholar 

  • Macpherson HL, Bergh Ø, Birkbeck TH (2012) An aerolysin-like enterotoxin from Vibrio splendidus, may be involved in intestinal tract damage and mortalities in turbot, Scophthalmus maximus, (L.), and cod, Gadus morhua L., larvae. J Fish Dis 35:153–167

    Article  CAS  PubMed  Google Scholar 

  • Mel SF, Fullner KJ, Wimermackin S, Lencer WI, Mekalanos JJ (2000) Association of protease activity in Vibrio cholerae vaccine strains with decreases in transcellular epithelial resistance of polarized T84 intestinal epithelial cells. Infection & Immunity 68:6487–6492

    Article  CAS  Google Scholar 

  • Montero D, Mathlouthi F, Tort L, Afonso JM, Torrecillas S, Fernándezvaquero A et al (2010) Replacement of dietary fish oil by vegetable oils affects humoral immunity and expression of pro-inflammatory cytokines genes in gilthead sea bream Sparus aurata. Fish & Shellfish Immunology 29:1073–1081

    Article  CAS  Google Scholar 

  • Murray MJ, Gonze MD, Nowak LR, Cobb CF (1994) Serum D(−)-lactate levels as an aid to diagnosing acute intestinal ischemia ☆. Am J Surg 167:575–578

    Article  CAS  PubMed  Google Scholar 

  • Muza-Moons MM, Schneeberger EE, Hecht GA (2004) Enteropathogenic Escherichia coli infection leads to appearance of aberrant tight junctions strands in the lateral membrane of intestinal epithelial cells. Cell Microbiology 6:783–793

    Article  CAS  Google Scholar 

  • Nielsen ME, Høi L, Schmidt AS, Qian D, Shimada T, Shen JY et al (2001) Is Aeromonas hydrophila the dominant motile Aeromonas species that causes disease outbreaks in aquaculture production in the Zhejiang Province of China? Dis Aquat Org 46:23–29

    Article  CAS  PubMed  Google Scholar 

  • Niklasson L (2013) Intestinal mucosal immunology of salmonids, response to stress and infection and crosstalk with the physical barrier.

  • O’Toole R, Hofsten JV, Rosqvist R, Olsson PE, Wolf-Watz H (2004) Visualisation of zebrafish infection by GFP-labelled Vibrio anguillarum. Microb Pathog 37:41–46

    Article  PubMed  Google Scholar 

  • Park JH, Ma L, Oshima T, Carter P, Coe L, Ma JW et al (2002) Polynitroxylated starch/TPL attenuates cachexia and increased epithelial permeability associated with TNBS colitis. Inflammation 26:1–11

    Article  CAS  PubMed  Google Scholar 

  • Parra D, Korytář T, Takizawa F, Sunyer JO (2016) B cells and their role in the teleost gut. Developmental & Comparative Immunology 64:150–166

    Article  CAS  Google Scholar 

  • Pastorelli L, De Salvo C, Mercado JR, Vecchi M, Pizarro TT (2013) Central role of the gut epithelial barrier in the pathogenesis of chronic intestinal inflammation: lessons learned from animal models and human genetics. Front Immunol 4:280

    Article  PubMed  PubMed Central  Google Scholar 

  • Rajasekaran SA, Barwe SP, Gopal J, Ryazantsev S, Schneeberger EE, Rajasekaran AK (2007) Na-K-ATPase regulates tight junction permeability through occludin phosphorylation in pancreatic epithelial cells. American Journal of Physiology Gastrointestinal & Liver Physiology 292:124–133

    Article  Google Scholar 

  • Ringø E, Olsen RE, Mayhew TM, Myklebust R (2003) Electron microscopy of the intestinal microflora of fish. Aquaculture 227:395–415

    Article  Google Scholar 

  • Ringø E, Jutfelt F, Kanapathippillai P, Bakken Y, Sundell K, Glette J, Glette J, Mayhew TM, Myklebust R, Olsen RE (2004) Damaging effect of the fish pathogen Aeromonas salmonicida ssp. salmonicida on intestinal enterocytes of atlantic salmon (Salmo salar L.) Cell Tissue Research 318:305–311

    Article  PubMed  Google Scholar 

  • Ringø E, Olsen RE, Myklebust R, Mayhew TM (2007) Bacterial translocation and pathogenesis in the digestive tract of larvae and fry. Aquaculture 268:251–264

    Article  Google Scholar 

  • Rochfort KD, Cummins PM (2015) Cytokine-mediated dysregulation of zonula occludens-1 properties in human brain microvascular endothelium. Microvasc Res 100:48–53

    Article  CAS  PubMed  Google Scholar 

  • Rombout JH, Abelli L, Picchietti S, Scapigliati G, Kiron V (2011) Teleost intestinal immunology. Fish & Shellfish Immunology 31:616–626

    Article  CAS  Google Scholar 

  • Salinas I, Esteban MA, Olsen RE, Meseguer J, Ringø E, Myklebust R (2008) In vitro, studies of Lactobacillus delbrueckii, subsp. Lactis in atlantic salmon (Salmo salar L.) foregut: tissue responses and evidence of protection against Aeromonas salmonicida subsp. salmonicida epithelial damage. Vet Microbiol 128:167–177

    Article  CAS  PubMed  Google Scholar 

  • Schroers V, Marel MVD, Neuhaus H, Steinhagen D (2009) Changes of intestinal mucus glycoproteins after peroral application of Aeromonas hydrophila, to common carp (Cyprinus carpio). Aquaculture 288:184–189

    Article  CAS  Google Scholar 

  • Scott GR, Rogers JT, Richards JG, Wood CM, Schulte PM (2004) Intraspecific divergence of ionoregulatory physiology in the euryhaline teleost Fundulus heteroclitus: possible mechanisms of freshwater adaptation. J Exp Biol 207(19):3399–3410

    Article  CAS  PubMed  Google Scholar 

  • Secombes CJ, Wang T, Hong S, Peddie S, Crampe M, Laing KJ et al (2001) Cytokines and innate immunity of fish. Developmental & Comparative Immunology 25:713–723

    Article  CAS  Google Scholar 

  • Song JY, Van MJ, Van Noorden CJ, Frederiks WF (1996) Redistribution of Ca2+, Mg2+-ATPase activity in relation to alterations of the cytoskeleton and tight junctions in hepatocytes of cholestatic rat liver. Eur J Cell Biol 71:277–285

    CAS  PubMed  Google Scholar 

  • Song XH, Zhao J, Bo YX, Liu ZJ, Wu K, Gong CL (2014) Aeromonas hydrophila, induces intestinal inflammation in grass carp (Ctenopharyngodon idella): an experimental model. Aquaculture 434:171–178

    Article  Google Scholar 

  • Spitz J, Yuhan R, Koutsouris A, Blatt C, Alverdy J, Hecht G (1995) Enteropathogenic Escherichia coli adherence to intestinal epithelial monolayers diminishes barrier function. Am J Physiol 268:374–379

    Google Scholar 

  • Sun X, Yang H, Nose K, Nose S, Haxhija EQ, Koga H et al (2008) Decline in intestinal mucosal IL-10 expression and decreased intestinal barrier function in a mouse model of total parenteral nutrition. Ajp Gastrointestinal & Liver Physiolog 294:G139–G147

    Article  CAS  Google Scholar 

  • Sun L, Liu S, Bao L, Yun L, Feng J, Liu Z (2015) Claudin multigene family in channel catfish and their expression profiles in response to bacterial infection and hypoxia as revealed by meta-analysis of RNA-seq datasets. Comparative Biochemistry & Physiology Part D Genomics & Proteomics 13:60–69

    Article  CAS  Google Scholar 

  • Sundh H (2009) Chronic stress and intestinal barrier function: implications for infection and inflammation in intensive salmon aquaculture. PhD thesis, University of Gothenburg, Sweden

  • Sundh H, Olsen RE, Fridell F, Gadan K, Evensen Ø, Glette J et al (2009) The effect of hyperoxygenation and reduced flow in fresh water and subsequent infectious pancreatic necrosis virus challenge in sea water, on the intestinal barrier integrity in atlantic salmon, Salmo salar l. J Fish Dis 32:687–698

    Article  CAS  PubMed  Google Scholar 

  • Syakuri H, Adamek M, Brogden G, Rakus KL, Matras M, Irnazarow I et al (2013) Intestinal barrier of carp (Cyprinus carpio L.) during a cyprinid herpesvirus 3-infection: molecular identification and regulation of the mRNA expression of claudin encoding genes. Fish & Shellfish Immunology 34:305–314

    Article  CAS  Google Scholar 

  • Syakuri H, Jung-Schroers V, Adamek M, Brogden G, Irnazarow I, Steinhagen D (2014) Beta-glucan feeding differentiated the regulation of mRNA expression of claudin genes and prevented an intestinal inflammatory response post Aeromonas hydrophila, intubation in common carp, Cyprinus carpio L. J Fish Dis 37:149–156

    Article  CAS  PubMed  Google Scholar 

  • Trinchet I, Djediat C, Huet H, Dao SP, Edery M (2011) Pathological modifications following sub-chronic exposure of medaka fish (Oryzias latipes) to microcystin-LR. Reprod Toxicol 32:329–340

    Article  CAS  PubMed  Google Scholar 

  • Ukena SN (2007) Probiotic, Escherichia coli, Nissle 1917 inhibits leaky gut by enhancing mucosal integrity. PLoS One 2:e1308

    Article  PubMed  PubMed Central  Google Scholar 

  • Urán PA, Schrama JW, Jhwm R, Obach A, Jensen L, Koppe W et al (2008) Soybean meal-induced enteritis in atlantic salmon (Salmo salar L.) at different temperatures. Aquac Nutr 14:324–330

    Article  Google Scholar 

  • Wen H, Feng L, Jiang W, Liu Y, Jiang J, Li S et al (2014) Dietary tryptophan modulates intestinal immune response, barrier function, antioxidant status and gene expression of tor and NRF2 in young grass carp (Ctenopharyngodon idella). Fish & Shellfish Immunology 40:275–287

    Article  CAS  Google Scholar 

  • Will C, Fromm M, Müller D (2007) Claudin tight junction proteins: novel aspects in paracellular transport. Peritoneal Dialysis International Journal of the International Society for Peritoneal Dialysis 28:577–584

    Google Scholar 

  • Wu Z, Milton D, Nybom P, Sjö A, Magnusson KE (1996) Vibrio cholerae hemagglutinin/protease (ha/protease) causes morphological changes in cultured epithelial cells and perturbs their paracellular barrier function. Microb Pathog 21:111–123

    Article  CAS  PubMed  Google Scholar 

  • Xu F (2013) Cloning and expression activity analysis of the tight junction protein genes of grass carp, Ctenopharyngodon idellus. Dissertation, University of Soochow

  • Yu H, Huang X, Ma Y, Min G, Ou W, Gao T et al (2013) Interleukin-8 regulates endothelial permeability by down-regulation of tight junction but not dependent on integrins induced focal adhesions. Int J Biol Sci 9:966–979

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang XJ, Yang WM, Zhang DF, Li TT, Gong XN, Li AH (2015) Does the gastrointestinal tract serve as the infectious route of Aeromonas hydrophila, in crucian carp (Carassius carassius)? Aquac Res 46:141–154

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Aquatic Animal Medicine, College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, China

    Wei-Guang Kong, Si-Si Li, Xiao-Xuan Chen, Yu-Qing Huang, Ying Tang & Zhi-Xin Wu

  2. Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Wuhan, 430070, China

    Wei-Guang Kong, Si-Si Li, Xiao-Xuan Chen, Yu-Qing Huang, Ying Tang & Zhi-Xin Wu

  3. Freshwater Aquaculture Collaborative Innovation Center, Wuhan, 430070, China

    Wei-Guang Kong, Si-Si Li, Xiao-Xuan Chen, Yu-Qing Huang, Ying Tang & Zhi-Xin Wu

Authors
  1. Wei-Guang Kong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Si-Si Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiao-Xuan Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yu-Qing Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ying Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Zhi-Xin Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhi-Xin Wu.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kong, WG., Li, SS., Chen, XX. et al. A study of the damage of the intestinal mucosa barrier structure and function of Ctenopharyngodon idella with Aeromonas hydrophila . Fish Physiol Biochem 43, 1223–1235 (2017). https://doi.org/10.1007/s10695-017-0366-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10695-017-0366-z

Keywords

Search

Navigation