Bacteriocinogeny in experimental pigs treated with indomethacin and Escherichia coli Nissle

Table 1 DNA primers used for polymerase chain reaction detection of colicin encoding genes

Bacteriocin type	Primer name	5'-sequence-3'	Length of PCR product (nt)
A	ColA-F	CGTGGGGAAAAGTCATCATC
	ColA-R	GCTTTGCTCTTTCCTGATGC	475
B	colicinB-F	AAGAAAATGACGAGAAGACG
	colicinB-R	GAAAGACCAAAGGCTATAAGG	493
D	ColD-F	CTGGACTGCTGCTGGTGATA
	ColD-R	GAAGGTGCGCCTACTACTGC	420
E1	colicinE1-F	TGTGGCATCGGGCGAGAATA
	colicinE1-R	CTGCTTCCTGAAAAGCCTTTT	650
E2	ColE2-F	TGATGCTGCTGCAAAAGAG
	ColE2-R	TTCAAAGCGTTCCCTACCAC	409
E3	ColE3-F	TAAGCAGGCTGCATTTGATG
	ColE3-R	TCGGATCTGGACCTTTCAAC	413
E4	ColE4-F	GAAGGCTGCATTTGATGCT
	ColE4-R	CGGATCCGGACCTTTAATTT	409
E5	ColE3-F	TAAGCAGGCTGCATTTGATG
	ColE5-R	TTGAATTCTCGAATCGTCCA	430
E6	ColE6-F	ACCGAACGTCCAGGTGTT
	ColE6-R	TTTAGCCTGTCGCTCCTGAT	399
E7	ColE7-F	GCATTCTGCCATCTGAAAT
	ColE7-R	CTTCTGCCCACTTTCTTTCG	431
E8	ColE3-F	TAAGCAGGCTGCATTTGATG
	ColE8-R	GACTGATTGGCTTGTCGTGA	449
E9	ColE3-F	TAAGCAGGCTGCATTTGATG
	ColE9-R	GACTTTTCTCCCTCCGACCT	418
Ia	ColIa-F	GCATGCAAATGACGCTCTTA
	ColIa-R	GAGGACGCCAGTTCTCTGTC	473
Ib	ColIb-F	AACGAGTGGGTCGATGATTC
	ColIb-R	CCTTTTCTGCGCTCGTATTC	464
Js	ColJs-F	TCAAAATGTTTGGGCTCCTC
	ColJs-R	TAATCTGCCCTGTCCCACTG	254
K	ColK-F	CAGAGGTCGCTGAACATGAA
	ColK-R	TCCGCTAAATCCTGAGCAAT	469
M	ColM-F	GCTTACCACTTCGCAAAACC
	ColM-R	GAGCGACTCTCCGATAATGC	429
N	ColN-F	AGCTTGGCGAGTATCTTGGA
	ColN-R	CAACACAGCCCCGAATAAAC	401
S4	ColS4-F	TATATGGCCCAACTGCTGGT
	ColS4-R	CGTAAGGACGGACACCTGTT	456
U	ColU-F	TGATTGCTGCGAGAAAAATG
	ColU-R	TCTGACAGCCTCTCCCTGTT	485
Y	ColY-F	GCAGGCAGAAAAGAACAAGG
	ColY-R	CGGACGTTATTTGCCTTCAT	477
5	Col5-F	CATTGGCAAAAGCGAAATCT
	Col5-R	TGCAACTCTGGAAACAATCG	443
10	Col10-F	GGTTACCGGATTTCCTGGAT
	Col10-R	TTCTAGATGCTTGGCCCACT	448

PCR: Polymerase chain reaction.

Table 2 Bacteriocinogeny of particular strains isolated at the end of experiment

Parameter	Small intestine			Colon and rectum
	Bacterio-cinogeny	Types of bacteriocin producers (No. of strains)	No. of unique bacteriocin producers	Bacterio-cinogeny	Types of bacteriocin producers (No. of strains)	No. of unique bacteriocin producers
Group A	22/55 (40%)	E1 (1); E1, Ia, V (1); E1, V (2); Ia (2); Ia, B, K, M, H47 (2); Ia, H47, V (2); Ia, V (6); Ib (3); J25 (1); S4, U (1); S4, V (1)	11	30/94 (32%)	B (1); B, H47, Ib, K, M (2); B, M (1); C7, E1, Ib, V (1); E1 (1); E1, Ia (1); E1, Ia, V (1); E1, V (2); E7 (1); H47, Ia, V (1); H47, S4 (1); H47, V (2); Ia (6); Ia, V (4); Ib (3); M (1); S4, V (1)	17
Group B	11/43 (26%)	E1 (7); Ia (2); Ia, V, H47 (1); Ib (1)	4	20/79 (25%)	B, H47, K, M, Ia (1); B, M (1); E1 (4); E1, V (1); Ia (11); Ia, V (1); Ib (1)	7
Group C	17/58 (29%)	B, M, V (1); Ia (1); Ia, V (7); Ib (6); S4 (2)	5	26/97 (27%)	B, Ia, (1); E1, Ib (1); J25, Ia (1); Ia (5); Ia, E7, V (1); Ia, H47 (1); Ia, M (1); Ia, V (8); Ib (7)	9
Group D	9/45 (20%)	E1 (3); E1, Ia (1); E1, Ia, V (1); E1, Ib (1); Ia (1); Ia, V (1); Ib (1)	7	20/89 (22%)	E1 (4); E1, Ia (1); E1, Ia, V (1); E1, Ib (2); H47, V (1); Ia (7); Ia, V (1); Ib (2); Ib, V (1)	9

Small intestine: Bacterial strains isolated from mucosa of the jejunum and ileum; Colon and rectum: Bacterial strains isolated from mucosa of the caecum, transverse colon and rectum; Group A: Control animals with no treatment; Group B: Probiotics alone (see text for details); Group C: Indomethacin alone (see text for details); Group D: Probiotics and indomethacin (see text for details); Bacteriocinogeny: Number of bacteriocinogenic strains out of all tested; Types: Particular bacteriocin types found in single isolates; M: Colicin M, not for microcin M.

Table 3 Analysis of porcine alveolar breath for hydrogen and methane (in ppm - parts per million) before and after the treatment

Group	Hydrogen before	Hydrogen after	Statistical significance	Methane before	Methane after	Statistical significance
A	N/A	3.50 ± 2.81	N/A	N/A	69.33 ± 56.64	N/A
B	6.0 ± 2.82	2.0 ± 0	NS	106.50 ± 94.05	80.00 ± 48.02	NS
C	1.17 ± 0.41	5.0 ± 3.29	NS	34.67 ± 25.65	66.17 ± 38.83	NS
D	2.0 ± 1.16	6.0 ± 6.0	NS	60.75 ± 34.77	62.00 ± 27.71	NS

Group A (controls with no treatment, n = 6), Group B (probiotics alone, n = 6), Group C (indomethacin alone, n = 6) and Group D (probiotics plus indomethacin, n = 6). N/A: Not applicable; NS: Not significant.