Potential of a fucoidan-rich Ascophyllum nodosum extract to reduce Salmonella shedding and improve gastrointestinal health in weaned pigs naturally infected with Salmonella

Venardou, Brigkita; O’Doherty, John V.; Maher, Shane; Ryan, Marion T.; Gath, Vivian; Ravindran, Rajeev; Kiely, Claire; Rajauria, Gaurav; Garcia-Vaquero, Marco; Sweeney, Torres

doi:10.1186/s40104-022-00685-4

Journal of Animal Science and Biotechnology

Table 3 Panel of target genes evaluated in the ileum and colon

From: Potential of a fucoidan-rich Ascophyllum nodosum extract to reduce Salmonella shedding and improve gastrointestinal health in weaned pigs naturally infected with Salmonella

Target gene	Accession No.	Forward primer 5’ to 3′ Reverse primer 5′ to 3’	Amplicon length, bp	Tm, °C
Immune response
IL1A	NM_214029.1	F: CAGCCAACGGGAAGATTCTG R: ATGGCTTCCAGGTCGTCAT	76	63.0 60.5
IL6	NM_214399.1	F: GACAAAGCCACCACCCCTAA R: CTCGTTCTGTGACTGCAGCTTATC	69	59.8 62.7
IL7	NM_214135.2	F: GAGTGACTATGGGCGGTGAGA R: GCGGGCGTGGTCATGA	63	61.8 56.9
CXCL8	NM_213867.1	F: TGCACTTACTCTTGCCAGAACTG R: CAAACTGGCTGTTGCCTTCTT	82	61.9 61.7
IL10	NM_214041.1	F: GCCTTCGGCCCAGTGAA R: AGAGACCCGGTCAGCAACAA	71	63.4 63.1
IL17A	NM_001005729.1	F: CCCTGTCACTGCTGCTTCTG R: TCATGATTCCCGCCTTCAC	57	60.6 60.4
IL22	XM_001926156.1	F: GATGAGAGAGCGCTGCTACCTGG R: GAAGGACGCCACCTCCTGCATGT	112	66.0 66.0
IFNG	NM_213948.1	F: TCTAACCTAAGAAAGCGGAAGAGAA R: TTGCAGGCAGGATGACAATTA	81	61.1 61.5
TNF	NM_214022.1	F: TGGCCCCTTGAGCATCA R: CGGGCTTATCTGAGGTTTGAGA	68	62.5 62.8
TGFB1	NM_214015.1	F: AGGGCTACCATGCCAATTTCT R: CGGGTTGTGCTGGTTGTACA	101	60.6 61.7
FOXP3	NM_001128438.1	F: GTGGTGCAGTCTCTGGAACAAC R: AGGTGGGCCTGCATAGCA	68	60.6 61.2
CCL20^a	NM_001024589.1	F: GCTCCTGGCTGCTTTGATG R: TTGCTTGCTGCTTCTGACTTG	66	58.8 57.9
TLR4	NM_001293317.1	F: TGCATGGAGCTGAATTTCTACAA R: GATAAATCCAGCACCTGCAGTTC	140	57·1 60·6
TP53^a	NM_213824.3	F: CCGGGTGGAAGGGAATTT R: CCACAACGCTGTGTCGAAAA	68	56.0 57.3
STAT3^a	NM_001044580	F: TCTTGAGAAGCCAATGGAGATTG R: TGGAGGAGGCGGGACTCT	69	58.9 60.5
Intestinal integrity
MUC2	AK231524	F: CAACGGCCTCTCCTTCTCTGT R: GCCACACTGGCCCTTTGT	70	63.1 62.1
TJP1/ZO-1	XM_005659811.1	F: TGAGAGCCAACCATGTCTTGAA R: CTCAGACCCGGCTCTCTGTCT	76	59.9 60.0
Cholinergic receptor
CHRM1^a	NM_214034.1	F: GCCATGGCCGCCTTCT R: GGTTCTCTGTCTCCCGGTAGATG	76	56.9 64.2
NADPH oxidases
NOX1^a	XM_003484140.3	F: CTTTGAAAGGATCCTCCGATTTT R: ATGGATACATGACCACCTTGGTAA	71	57.1 59.3
DUOX2^a	NM_213999.2	F: CTGGGCCTTGACATAGATGAGAT R: GGCAAAAAGGTGTCTGAAGAAGA	108	60.6 58.9
Reference genes
PPIA	NM_214353.1	F: CGGGTCCTGGCATCTTGT R: TGGCAGTGCAAATGAAAAACT	75	62.1 60.7
B2M	NM_213978.1	F: CGGAAAGCCAAATTACCTGAAC R: TCTCCCCGTTTTTCAGCAAAT	83	58.2 58.4
GAPDH	AF017079.1	F: CAGCAATGCCTCCTGTACCA R: ACGATGCCGAAGTTGTCATG	72	62.2 62.1

bp, Base pairs; Tm, Melting temperature; IL1A, Interleukin 1 alpha; IL6, Interleukin 6; IL7, Interleukin 7; CXCL8, C-X-C motif chemokine ligand 8; IL10, Interleukin 10; IL17A, Interleukin 17 alpha; IL22, Interleukin 22; IFNG, Interferon gamma; TNF, Tumour necrosis factor; TGFB1, Transforming growth factor beta 1; FOXP3, Forkhead box P3; CCL20, C-C motif chemokine ligand 20; TLR4, Toll-like receptor 4; TP53, Tumour protein p53; STAT3, Signal transducer and activator of transcription 3; MUC2, Mucin 2; TJP1/ZO-1, Tight junction protein 1/Zona occludens 1; CHRM1, Cholinergic receptor muscarinic 1; NOX1, Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 1; DUOX2, Dual oxidase 2; PPIA, Peptidylprolyl isomerase A; B2M, Beta-2-microglobulin; GAPDH, Glyceraldehyde-3-phosphate dehydrogenase
^aGenes encoding proteins with an established role in facilitating or inhibiting Salmonella infection. These proteins are associated with chemotaxis (CCL20), production of reactive oxygen species (NOX1, DUOX2) anti-inflammatory activity (STAT3, CHRM1) and cell survival and death (TP53) [54,55,56,57,58,59]

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ISSN: 2049-1891

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