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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
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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

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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

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Supplementary material 1 (TIFF 415 kb)
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Supplementary material 3 (TIFF 433 kb)
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Supplementary material 4 (TIFF 216 kb)
13205_2019_1956_MOESM5_ESM.rtf (136 kb)
Supplementary material 5 (RTF 135 kb)

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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

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