Background
Methods
Animal and samplings
Dog no. | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Gendera
| M | F | M | M | M | F | F | M | F | F | F | F | M | M | M | |
Ageb
| 1 | 3 | 1 | 2 | 3 | 1 | 2 | 3 | 3 | 1 | 3 | 1 | 2 | 1 | 3 | |
Clinical signsc
| MD, W,A | H | MD, W,A | H | MD, W,A | MD, W,A | H | H | H | MD, W,A | H | H | MD, W,A | H | H | |
Persistence of Helicobacter spirochetes | Mucosal surface | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + |
Crypt | + | + | + | - | - | + | - | - | - | + | - | + | + | - | - | |
Helicobacter spirochete colonization statusd
| +3 | +2 | +2 | +1 | +1 | +2 | +1 | +1 | +1 | +3 | +1 | +2 | +1 | +2 | +1 | |
Inflammatory cell Infiltratione
| LP | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + |
N | + | + | + | - | - | + | - | - | - | + | - | + | + | - | - | |
E | + | - | + | - | - | - | - | - | - | + | - | - | - | - | - | |
M | - | + | + | - | - | + | + | - | + | + | - | - | + | + | + | |
Lymphoid folliclesf
| +2 | +2 | +2 | 0 | 0 | +3 | +1 | 0 | +1 | +3 | 0 | +2 | +1 | +1 | +1 | |
Morphological changes | Surface epithelial injury | + | + | + | - | + | + | - | - | + | + | - | + | + | + | - |
Crypt hyperplasia | - | - | + | - | + | - | + | - | + | + | - | - | - | - | + | |
Crypt dilation/distortion | + | - | + | - | + | - | + | - | + | + | - | - | - | - | - | |
Mucosal fibrosis and atrophy | + | + | + | - | - | + | + | - | - | + | - | + | - | + | - | |
Severity of inflammatory and morphological changesg
| Mi | Mo | Ma | No | Mi | Ma | Mi | No | Mi | Ma | No | Mo | Mi | Mo | Mi |
Polymerase chain reaction (PCR) amplification
Dog No. | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Helicobacteraceae (H276f/H676r) | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + |
H.bilis (P17f/P17r) | - | - | + | - | + | + | + | + | + | - | + | - | - | - | - |
H.bizzozeronii (Bi1F/Bi2R) | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + |
H.canis (canis3F/canis4R) | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + |
H.felis (Fe1F/Fe3R) | - | - | + | - | - | + | - | + | + | - | + | - | - | - | + |
H.pylori(HP-for/HP-REV) | - | - | - | - | - | - | - | - | - | - | + | - | - | - | + |
H.salomonis(HSALS/HSALR) | + | + | + | - | + | + | + | - | + | - | + | + | + | + | + |
Primera
| Sequence (5' - 3')b
| Product size (bp) | PCR conditions | Target sequence | Referencesc
|
---|---|---|---|---|---|
H276f | CTA TGA CGG GTA TCC GGC | 374 |
94°c for 15 min and 30 s,52°c for 30 s,72°c for 30 s, 35 cycles,72°c for 7 min
|
Helicobacter spp.16S rRNA | Riley et al. [17] |
H676r | ATT CCA CCT ACC TCT CCC A | ||||
p17f | ATG GAA CAG ATA AAG ATT TTA AAG CAA CTT CAG | 435 |
94°c for 15 min and 30 s,55°c for 30 s,72°c for 30 s, 35 cycles,72°c for 7 min
|
H. bilis specifict protein | Feng et al. [21] |
p17r | CTA TGC AAG TTG TGC GTT AAG CAT | ||||
Fe1F | TTT GGT GCT CAC TAA CGC CCT C | 434 (438) |
94°c for 15 min and 30 s,52°c for 30 s,72°c for 30 s, 35 cycles,72°c for 7 min
|
H. felis urease gene | Baele et al. [19] |
Fe3R | TTC AAT CTG ATC GCG TAA AG | ||||
Bi1F | AAC CAA Y AG CCC CAG CAG CC | 373 |
94°c for 15 min and 30 s,54°c for 30 s,72°c for 30 s, 35 cycles,72°c for 7 min
|
H. bizzozeronii urease gene | Baele et al. [19] |
Bi2R | TGG TTT TAA GGT TCC AGC GC | ||||
HSALF | CAT TTT CAA AGA GGG CTT GC | 537 |
94°c for 15 min and 30 s,51°c for 30 s,72°c for 30 s, 35 cycles,72°c for 7 min
|
H. salomonis HSP60 | Mikkonen et al. [20] |
HSALR | GCA CAC CCC TCA GTT TGT TT | ||||
HP-FOR | TTA TCG GTA AAG ACA CCA GAA A | 132 |
94°c for 15 min and 30 s,49°c for 30 s,72°c for 30 s, 35 cycles,72°c for 7 min
|
H. pylori urease C gene | He et al. [22] |
HP-REV | ATC ACA GCG CAT GTC TTC | ||||
canis3Fd
| TAA GCG CGG TAT GGA TAA GG | 254 |
94°c for 15 min and 30 s,52°c for 30 s,72°c for 30 s, 35 cycles,72°c for 7 min
|
H. canis HSP60 | EU233451 |
canis4Rd
| TTA AGT AGC CGC GGT CAA AC |