Advertisement

3 Biotech

, 9:422 | Cite as

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

  • Chandrakanta Rawat
  • Nihar Ranjan SahooEmail author
  • Shivaji S. Wagh
  • Pushpendra Kumar
  • Subodh Kumar
  • Arvind Sonwane
  • Salauddin Qureshi
  • Amit Kumar
  • Manjit Panigrahi
Original Article
  • 28 Downloads

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.

Keywords

Native pig E. coli TFRC ACK1 MAT RFLP qPCR 

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

Notes

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.

Compliance with ethical standards

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.

Supplementary material

13205_2019_1956_MOESM1_ESM.tif (416 kb)
Supplementary material 1 (TIFF 415 kb)
13205_2019_1956_MOESM2_ESM.tif (473 kb)
Supplementary material 2 (TIFF 473 kb)
13205_2019_1956_MOESM3_ESM.tif (434 kb)
Supplementary material 3 (TIFF 433 kb)
13205_2019_1956_MOESM4_ESM.tif (216 kb)
Supplementary material 4 (TIFF 216 kb)
13205_2019_1956_MOESM5_ESM.rtf (136 kb)
Supplementary material 5 (RTF 135 kb)

References

  1. Baker DR, Billey LO, Francis DH (1997) Distribution of K88 Escherichia coli-adhesive and nonadhesive phenotypes among pigs of four breeds. Vet Microbiol 54:123–132CrossRefGoogle Scholar
  2. 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 37:891–894PubMedPubMedCentralGoogle Scholar
  3. 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 59:203–212CrossRefGoogle Scholar
  4. 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-NatalGoogle Scholar
  5. 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 CrossRefPubMedPubMedCentralGoogle Scholar
  6. Dekker J, Rossen JW, Büller HA, Einerhand AW (2002) The MUC family: an obituary. Trends Biochem Sci 27:126–131CrossRefGoogle Scholar
  7. 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 26:237–242CrossRefGoogle Scholar
  8. 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, GermanyGoogle Scholar
  9. 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 24:393–396CrossRefGoogle Scholar
  10. 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:e24202CrossRefGoogle Scholar
  11. 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 41:21–25CrossRefGoogle Scholar
  12. 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 4:225CrossRefGoogle Scholar
  13. 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-A2Google Scholar
  14. Li YH, Qiu XT, Li HJ, Zhang Q (2007) Adhesive patterns of Escherichia coli F4 in piglets of three breeds. J Genet Genomics 34:591–599CrossRefGoogle Scholar
  15. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408CrossRefGoogle Scholar
  16. 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 52:129–136CrossRefGoogle Scholar
  17. 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 11:557–560CrossRefGoogle Scholar
  18. 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 CrossRefGoogle Scholar
  19. 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 33:441–447CrossRefGoogle Scholar
  20. 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):5–14CrossRefGoogle Scholar
  21. Reich DE (2001) Linkage disequilibrium in the human genome. Nature 411:199–204CrossRefGoogle Scholar
  22. 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 126:30–36CrossRefGoogle Scholar
  23. 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 55:333–338Google Scholar
  24. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbour Laboratory Press, New York, pp 527–535Google Scholar
  25. 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 43:324–327CrossRefGoogle Scholar
  26. 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 152:87–92CrossRefGoogle Scholar
  27. 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 537–549Google Scholar
  28. 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 8:405–411CrossRefGoogle Scholar
  29. 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 75:196–198Google Scholar
  30. 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 46:107–111CrossRefGoogle Scholar
  31. 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 61:321–328CrossRefGoogle Scholar
  32. 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 51:1307–1320CrossRefGoogle Scholar
  33. Takezaki N, Nei M (1996) Genetic distance and reconstruction of phylogenetic trees from microsatellite DNA. Genetics 144:389–399PubMedPubMedCentralGoogle Scholar
  34. 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 147:96–103CrossRefGoogle Scholar
  35. Van den Broeck W, Cox E, Oudega B, Goddeeris BM (2000) The F4 fimbrial antigen of Escherichia coli and its receptors. Vet Microbiol 71:223–244CrossRefGoogle Scholar
  36. 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 93:297–303CrossRefGoogle Scholar
  37. 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 2:10CrossRefGoogle Scholar
  38. 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 124:225–229CrossRefGoogle Scholar
  39. 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 58:1112–1117CrossRefGoogle Scholar
  40. 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 274:8524–8530CrossRefGoogle Scholar

Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  • Chandrakanta Rawat
    • 1
  • Nihar Ranjan Sahoo
    • 1
    • 2
    Email author
  • Shivaji S. Wagh
    • 1
  • Pushpendra Kumar
    • 1
  • Subodh Kumar
    • 1
  • Arvind Sonwane
    • 1
  • Salauddin Qureshi
    • 3
  • Amit Kumar
    • 1
  • Manjit Panigrahi
    • 1
  1. 1.Central Instrumentation Facility, Division of Animal GeneticsICAR-Indian Veterinary Research InstituteIzatnagar, BareillyIndia
  2. 2.Swine Production FarmICAR-Indian Veterinary Research InstituteIzatnagar, BareillyIndia
  3. 3.Division of Biological StandardizationICAR-Indian Veterinary Research InstituteIzatnagarIndia

Personalised recommendations