Skip to main content
SpringerLink
Log in

Association of ACK1, TFRC polymorphism with diarrhoeagenic E. coli adhesion patterns and their jejunal expression profile in Indian Ghurrah pigs

  • Original Article
  • Published:
3 Biotech Aims and scope Submit manuscript

Abstract

A total of 9 SNPs located in TFRC and ACK1 genes of SSC13q41 genomic region were examined for their association with the adhesion pattern of native Indian pigs using local isolate of diarrhoeagenic E. coli. Phenotypic evaluation of adhesion pattern of 150 pigs revealed 116 animals positive for adhesion, whereas 34 animals had non-adhesive phenotype. Among the adhesive animals, 6, 87 and 23 pigs were strongly adhesive, weakly adhesive and adhesive, respectively. PCR–RFLP study revealed 8 polymorphic SNPs with low to moderate PIC ranging from 7.39 to 37.25% and low to high heterozygosities (8–70%). The loci g.291 C > T, rs81218930 C > T, rs318751568 C > T of TFRC and g.93222 C > A g.94600 C > T of ACK1 showed significant departure from HWE. The genotypic frequencies of the SNPs as well as the haplotypes did not differ significantly (P > 0.05) across the adhesion patterns except one SNP (ACK1-g.107371 A > C). Among the g.107371 A > C genotypes observed, CA was associated with non-adhesive phenotype. Furthermore, TFRC mRNA expression levels were found to be significantly (P < 0.05) different among various adhesive phenotypes, whereas that of ACK1 was significantly (P < 0.05) different between non-adhesive and adhesive groups. The significant association of SNP (ACK1-g.107371 A > C), which was also previously reported to influence ETECF4 mediated diarrhoea susceptibility, implicates its wider application in genetic control of piglet diarrhoea. Furthermore, the up-regulation of TFRC gene expression in adhesive group supports its proposed role in activation of immune cells against E. coli and intracellular iron transport.

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

Similar content being viewed by others

Abbreviations

ACK1:

Activated CDC42 kinase 1

AD:

Allelic diversity

AICRP:

All India coordinated research project

AT:

Annealing temperature

BHI:

Brain–heart infusion

CI:

Confidence interval

dbSNP:

Single-nucleotide polymorphism database

DT:

Digestion temperature

EDTA:

Ethylenediaminetetraacetic acid

ETECF4:

F4 fimbriae of enterotoxigenic Escherichia coli

Ht:

Heterozygosity

HWE:

Hardy Weinberg equilibrium (χ2-probability)

LB:

Lysogeny broth

LD:

Linkage disequilibrium

MAT:

Microscopic adhesion test

MUC13:

Mucin 13

MUC20:

Mucin 20

MUC4:

Mucin 4

N:

Number of observations

OD:

Optical density

PBS:

Phosphate-buffered saline

PIC:

Polymorphism information content

RE:

Restriction enzyme

RFLP:

Restriction fragment length polymorphism

RT:

Reverse transcription

SNP:

Single-nucleotide polymorphism

SSC:

Sus scrofa (pig)

TFRC:

Transferrin receptor

References

  • Baker DR, Billey LO, Francis DH (1997) Distribution of K88 Escherichia coli-adhesive and nonadhesive phenotypes among pigs of four breeds. Vet Microbiol 54:123–132

    Article  CAS  Google Scholar 

  • Bijlsma IG, de Nijs A, van der Meer C, Frik JF (1982) Different pig phenotypes affect adherence of Escherichia coli to jejunal brush borders by K88ab, K88ac, or K88ad antigen. Infect Immun 37:891–894

    CAS  PubMed  PubMed Central  Google Scholar 

  • Billey LO, Erickson AK, Francis DH (1998) Multiple receptors on porcine intestinal epithelial cells for the three variants of Escherichia coli K88 fimbrial adhesin. Vet Microbiol 59:203–212

    Article  CAS  Google Scholar 

  • Chaora N (2013) Breed susceptibility to enterotoxigenic and enteroaggragative Escherichia coli strains in South African pigs. Thesis, Master of Science in Agriculture (Animal Science), University of KwaZulu-Natal

  • Dai C, Sun L, Xia R, Sun S, Zhu G, Wu S, Bao W (2017) Correlation between the methylation of the FUT1 promoter region and FUT1 expression in the duodenum of piglets from newborn to weaning. 3 Biotech 7:247. https://doi.org/10.1007/s13205-017-0880-9

    Article  PubMed  PubMed Central  Google Scholar 

  • Dekker J, Rossen JW, Büller HA, Einerhand AW (2002) The MUC family: an obituary. Trends Biochem Sci 27:126–131

    Article  CAS  Google Scholar 

  • Edfors-Lilja I, Gustafsson U, Duval-Iflah Y, Ellergren H, Jo-hansson M, Juneja RK, Marklund L, Andersson L (1995) The porcine intestinal receptor for Escherichia coli K88ab, K88ac: regional localization on chromosome 13 and influence of IgG response to the K88 antigen. Anim Genet 26:237–242

    Article  CAS  Google Scholar 

  • Eliades NG, Eliades DG (2009). HAPLOTYPE ANALYSIS: software for analysis of haplotypes data. Distributed by the authors. Forest Genetics and Forest Tree Breeding, Georg-Augst University Goettingen, Germany

  • Guerin G, Duval-Iflah Y, Bonneau M, Bertaud M, Guillaume P, Ollivier L (1993) Evidence for linkage between K88ab, K88ac intestinal receptors to Escherichia coli and transferrin loci in pigs. Anim Genet 24:393–396

    Article  CAS  Google Scholar 

  • Harel E, Rubinstein A, Nissan A, Khazanov E, Milbauer MN, Barenholz Y, Tirosh B (2011) Enhanced transferrin receptor expression by proinflammatory cytokines in enterocytes as a means for local delivery of drugs to inflamed gut mucosa. PLoS ONE 6:e24202

    Article  CAS  Google Scholar 

  • Jacobsen M, Kracht SS, Esteso G, Cirera S, Edfors I, Archibald A, Bendixen C, Andersson L, Fredholm M, Jorgensen CB (2009) Refined candidate region specified by haplotype sharing for Escherichia coli F4ab/F4ac susceptibility alleles in pigs. Anim Genet 41:21–25

    Article  Google Scholar 

  • Jacobsen M, Cirera S, Joller D, Esteso G, Kracht SS, Edfors I, Bendixen C, Archibald AL, Vogeli P, Neuenschwander S, Bertschinger HU, Rampoldi A, Andersson L, Fredholm M, Jorgensen CB (2011) Characterisation of five candidate genes within the ETEC F4ab/ac candidate region in pigs. BMC Res Notes 4:225

    Article  CAS  Google Scholar 

  • Jorgensen CB, Cirera S, Archibald AL, Anderson L, Fredholm M, Edfors-Lilja I (2004) Porcine polymorphisms and methods for detecting them. In: International application published under the patent cooperation treaty (PCT), PCT/DK2003/000807; WO:2004/048606-A2

  • Li YH, Qiu XT, Li HJ, Zhang Q (2007) Adhesive patterns of Escherichia coli F4 in piglets of three breeds. J Genet Genomics 34:591–599

    Article  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408

    Article  CAS  Google Scholar 

  • Meijerink E, Neuenschwander S, Fries R, Dinter A, Bertschinger HU, Stranzinger G, Vogeli P (2000) A DNA polymorphism influencing alpha(l,2) fucosyltransferase activity of the pig FUT1 enzyme determines susceptibility of small intestinal epithelium to Escherichia coli F18 adhesion. Immunogenetics 52:129–136

    Article  CAS  Google Scholar 

  • Moon HW, Hoffman LJ, Cornick NA (1999) Prevalences of some virulence genes among Escherichia coli isolates from swine presented to a diagnostic laboratory in Iowa. J Vet Diagn Invest 11:557–560

    Article  CAS  Google Scholar 

  • Nygard AB, Jorgenson CB, Cierra S, Fredholm M (2007) Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR. BMC Molecular Biology 8:67. https://doi.org/10.1186/1471-2199-8-67

    Article  Google Scholar 

  • Python P, Jorg H, Neuenschwander S, Hagger C, Stricker C, Burgi E, Bertschinger HU, Stranzinger G, Vogeli P (2002) Fine-mapping of the intestinal receptor locus for enterotoxi-genic Escherichia coli F4ac on porcine chromosome 13. Anim Genet 33:441–447

    Article  CAS  Google Scholar 

  • Python P, Jorg H, Neuenschwander S, Asai-Coakwell M, Hagger C, Burgi E, Bertschinger HU, Stranzinger G, Vogeli P (2005) Inheritance of the F4ab, F4ac and F4ad E. coli receptors in swine and examination of four candidate genes for F4acR. J Anim Breed Genet 122(S1):5–14

    Article  CAS  Google Scholar 

  • Reich DE (2001) Linkage disequilibrium in the human genome. Nature 411:199–204

    Article  CAS  Google Scholar 

  • Ren J, Tang H, Yan XM, Huang X, Zhang B, Ji HY, Huang LS (2009) A pig-human comparative RH map comprising 20 genes on pig chromosome 13q41 that harbors the ETEC F4ac receptor locus. J Anim Breed Genet 126:30–36

    Article  CAS  Google Scholar 

  • Rita DM, Wbipp SC, Max FR (1994) Resistance of Chinese Meishan, Fengjing, and Minzhu pigs to the K88ac + strain of Escherichia coli. Am J Vet Res 55:333–338

    Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbour Laboratory Press, New York, pp 527–535

    Google Scholar 

  • Schroyen M, Stinckens A, Verhelst R, Cox E, Niewold T, Buys N (2012) Susceptibility of piglets to enterotoxigenic E. coli is not related to the expression of MUC13 and MUC20. Anim Genet 43:324–327

    Article  CAS  Google Scholar 

  • Schroyen M, Stinckens A, Verhelst R, Janssens S, Cox E, Goddeeris BM, Georges M, Niewold T, Buys N (2013) The effect of enterotoxigenic Escherichia coli F4ab/ac on early-weaned piglets: a gene expression study. Vet Immunol Immunopathol 152:87–92

    Article  CAS  Google Scholar 

  • Sellwood R (1980) Genetic susceptibility to intestinal infection-animal models. In: Rotter JI, Samloff IM, Rimoin DL (eds) The genetics and heterogeneity of common gastrointestinal disorders. Academic Press, London, pp 537–549

  • Sellwood R, Gibbons RA, Jones GW, Rutter JM (1975) Adhesion of enteropathogenic Escherichia coli to pig intestinal brush borders: the existence of two pig phenotypes. J Med Microbiol 8:405–411

    Article  CAS  Google Scholar 

  • Shome R, Shome BR, Rahman M, Kumar A, Murugkar HV, Rahman H, Bujarbaruah K (2005) Plasmid diversity in Escherichia coli strains isolated from piglet diarrhoea. Indian J Anim Sci 75:196–198

    CAS  Google Scholar 

  • Sinha R, Sahoo NR, Kumar P, Qureshi S, Kumar A, Ravikumar GVPPS, Bhushan B (2018a) Comparative jejunal expression of MUC 13 in indian pigs differentially adhesive to diarrhogenic E. coli. J Appl Anim Res 46:107–111

    Article  CAS  Google Scholar 

  • Sinha R, Sahoo NR, Shrivastava K, Kumar P, Qureshi S, Kumar A, Ravikumar GVPPS, Bhushan B (2018b) Effect of Mucin 13 gene polymorphism on diarrhoeagenic E. coli adhesion pattern and its expression analysis in native Indian pigs. Archiv Anim Breed 61:321–328

    Article  Google Scholar 

  • Sinha R, Sahoo NR, Shrivastava K, Kumar P, Qureshi S, Kumar A, Ravikumar GVPPS, Bhushan B (2019) Resistance to ETEC F4/F18 mediated piglet diarrhoea: opening the gene black box. Trop Anim Health Prod 51:1307–1320

    Article  Google Scholar 

  • Takezaki N, Nei M (1996) Genetic distance and reconstruction of phylogenetic trees from microsatellite DNA. Genetics 144:389–399

    CAS  PubMed  PubMed Central  Google Scholar 

  • Trevisi P, Corrent E, Messori S, Formica S, Priori D, Bosi P (2012) Supplementary tryptophan downregulates the expression of genes induced by the gut microbiota in healthy weaned pigs susceptible to enterotoxigenic Escherichia coli F4. Livest Sci 147:96–103

    Article  Google Scholar 

  • Van den Broeck W, Cox E, Oudega B, Goddeeris BM (2000) The F4 fimbrial antigen of Escherichia coli and its receptors. Vet Microbiol 71:223–244

    Article  Google Scholar 

  • Van Poucke M, Yerle M, Tuggle C, Piumi F, Genet C, Van Zeveren A, Peelman LJ (2001) Integration of porcine chromosome 13 maps. Cytogenet Cell Genet 93:297–303

    Article  Google Scholar 

  • Vu Khac H, Holoba E, Pilipcinec E, Blanco M, Blanco JE, Mora A (2006) Serotypes, virulence genes and PFGE profiles of Escherichia E. coli isolated from pigs with post-weaning diarrhoea in Slovakia. BMC Vet Res 2:10

    Article  Google Scholar 

  • Wang YZ, Ren J, Lan LT, Yan XM, Huang X, Peng QL, Tang H, Zhang B, Ji HY, Huang LS (2007) Characterization of polymorphisms of transferrin receptor and their association with susceptibility to ETEC F4ab/ac in pigs. J Anim Breed Genet 124:225–229

    Article  CAS  Google Scholar 

  • Yan X, Huang X, Ren J, Zou Z, Yang S, Ouyang J, Zeng W, Yang B, Xiao S, Huang L (2009) Distribution of Escherichia coli F4 adhesion phenotypes in pigs of 15 Chinese and Western breeds and a White Duroc 9 Erhualian intercross. J Med Microbiol 58:1112–1117

    Article  CAS  Google Scholar 

  • Yang W, Lin Q, Guan JL, Cerione RA (1999) Activation of the Cdc42-associated tyrosine kinase-2 (ACK-2) by cell adhesion via integrin β1. J Biol Chem 274:8524–8530

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are thankful to the Director, IVRI, Izzatnagar, Bareilly, for providing necessary facilities to carry out this research work. Furthermore, they are thankful to owners of different slaughter houses in and around Bareilly for facilitating collection of samples.

Funding

The authors are thankful to Indian Council of Agricultural Research (ICAR) for providing necessary financial assistance in the form of an institute project (Grant No. IXX10577) to corresponding author as well as institute fellowship to first author during the study.

Author information

Author notes

  1. Chandrakanta Rawat and Nihar Ranjan Sahoo have contributed equally.

Authors and Affiliations

  1. Central Instrumentation Facility, Division of Animal Genetics, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP, 243122, India

    Chandrakanta Rawat, Nihar Ranjan Sahoo, Shivaji S. Wagh, Pushpendra Kumar, Subodh Kumar, Arvind Sonwane, Amit Kumar & Manjit Panigrahi

  2. Swine Production Farm, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP, 243122, India

    Nihar Ranjan Sahoo

  3. Division of Biological Standardization, ICAR-Indian Veterinary Research Institute, Izatnagar, India

    Salauddin Qureshi

Authors
  1. Chandrakanta Rawat
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nihar Ranjan Sahoo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shivaji S. Wagh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Pushpendra Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Subodh Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Arvind Sonwane
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Salauddin Qureshi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Amit Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Manjit Panigrahi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nihar Ranjan Sahoo.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Rawat, C., Sahoo, N.R., Wagh, S.S. et al. Association of ACK1, TFRC polymorphism with diarrhoeagenic E. coli adhesion patterns and their jejunal expression profile in Indian Ghurrah pigs. 3 Biotech 9, 422 (2019). https://doi.org/10.1007/s13205-019-1956-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s13205-019-1956-5

Keywords

Search

Navigation