Background
Methods
Bacterial isolation and identification
Antimicrobial susceptibility testing
PCR amplification of ARGs
Target | Primer sequence (5′ to 3′) | Amplicon size (bp) | Reference |
---|---|---|---|
β-lactamase | |||
bla
CTX-M-1
| F: GGTTAAAAAATCACTGCGTC | 873 | Cui et al., 2015 [16] |
R: TTACAAACCGTCGGTGACGA | |||
bla
CTX-M-2
| F: CGACGCTACCCCTGCTATT | 552 | Zong et al., 2008 [17] |
R: CCAGCGTCAGATTTTTCAGG | |||
bla
CTX-M-8
| F: TCGCGTTAAGCGGATGATGC | 689 | Zong et al., 2008 [17] |
R: AACCCACGATGTGGGTAGC | |||
bla
CTX-M-9
| F: AGAGTGCAACGGATGATG | 868 | Cui et al., 2015 [16] |
R: CCAGTTACAGCCCTTCGG | |||
bla
CTX-M-25
| F: TTGTTGAGTCAGCGGGTTGA | 497 | Liu et al., 2015 [18] |
R: GCGCGACCTTCCGGCCAAAT | |||
bla
SHV
| F: CGCCGGGTTATTCTTATTTGTCGC | 1015 | Zong et al., 2008 [17] |
R: TCTTTCCGATGCCGCCGCCAGTCA | |||
bla
TEM
| F: ATGAGTATTCAACTTTCCG | 876 | This study |
R: CCAATGCTTAATCAGTGAG | |||
bla
OXA
| F: ATTAAGCCCTTTACCAAACCA | 890 | Cui et al., 2015 [16] |
R: AAGGGTTGGGCGATTTTGCCA | |||
bla
MOX
| F: GCTGCTCAAGGAGCACAGGAT | 520 | Cui et al., 2015 [16] |
R: CACATTGACATAGGTGTGGTGC | |||
bla
FOX
| F: AACATGGGGTATCAGGGAGATG | 190 | Cui et al., 2015 [16] |
R: CAAAGCGCGTAACCGGATTGG | |||
bla
DHA
| F: AACTTTCACAGGTGTGCTGGGT | 405 | Cui et al., 2015 [16] |
R: CCGTACGCATACTGGCTTTGC | |||
bla
CIT
| F: TGGCCAGAACTGACAGGCAAA | 462 | Cui et al., 2015 [16] |
R: TTTCTCCTGAACGTGGCTGGC | |||
bla
ACC
| F: AACAGCCTCAGCAGCCGGTTA | 346 | Cui et al., 2015 [16] |
R: TTCGCCGCAATCATCCCTAGC | |||
bla
MIR
| F: TCGGTAAAGCCGATGTTGCGG | 302 | Cui et al., 2015 [16] |
R: CTTCCACTGCGGCTGCCAGTT | |||
PMQRs | |||
qnrA
| F: ATTTCTCACGCCAGGATTTG | 516 | Colobatiu et al.,2015 [19] |
R: GATCGGCAAAGGTTAGGTCA | |||
qnrB
| F: GATCGTGAAAGCCAGAAAGG | 476 | Colobatiu et al.,2015 [19] |
R: ACGATGCCTGGTAGTTGTCC | |||
qnrD
| F: CGAGATCAATTTACGGGGAATA | 656 | Cui et al.,2015 [13] |
R: AACAAGCTGAAGCGCCTG | |||
qnrS
| F: ACGACATTCGTCAACTGCAA | 417 | Colobatiu et al.,2015 [19] |
R: TAAATTGGCACCCTGTAGGC | |||
aac(6′)-Ib-cr
| F: CCCGCTTTCTCGTAGCA | 544 | Colobatiu et al.,2015 [19] |
R: TTAGGCATCACTGCGTCTTC | |||
qepA
| F: CGTGTTGCTGGAGTTCTTC | 403 | Colobatiu et al.,2015 [19] |
R: CTGCAGGTACTGCGTCATG | |||
QRDR | |||
gyrA
| F: TACACCGGTCAACATTGAGG | 648 | Hu et al.,2007 [20] |
R: TTAATGATTGCCGCCGTCGG | |||
gyrB
| F: TGAAATGACCCGCCGTAAAGG | 309 | Hu et al.,2007 [20] |
R: GCTGTGATAACGCAGTTTGTCCGGG | |||
parC
| F: GTACGTGATCATGGACCGTG | 531 | Hu et al.,2007 [20] |
R: TTCGGCTGGTCGATTAATGC | |||
parE
| F: ATGCGTGCGGCTAAAAAAGTG | 290 | Hu et al.,2007 [20] |
R: TCGTCGCTGTCAGGATCGATAC |
PFGE
Results
Bacterial isolation and identification
Strain name | Serotype | Isolation year | Origin | Province |
---|---|---|---|---|
TYSF1412001 | 2a | 2014 | Yak | Tibet |
GBSF1412056 | 2a | 2014 | Beef cattle | Gansu |
GBSF1501026 | 2a | 2015 | Beef cattle | Gansu |
GBSF1501071 | Xv | 2015 | Beef cattle | Gansu |
GYSF1501076 | 6 | 2015 | Yak | Gansu |
QYSF1501088 | 6 | 2015 | Yak | Qinghai |
XBSF1501093 | 2b | 2015 | Beef cattle | Xinjiang |
GBSF1501105 | 2a | 2015 | Beef cattle | Gansu |
SBSF1501123 | 4a | 2015 | Beef cattle | Shanxi |
QYSF1502130 | 6 | 2015 | Yak | Qinghai |
GBSF1502176 | 2a | 2015 | Beef cattle | Gansu |
GYSF1502197 | 6 | 2015 | Yak | Gansu |
SBSF1502219 | 4a | 2015 | Beef cattle | Shanxi |
XBSF1502236 | 2b | 2015 | Beef cattle | Xinjiang |
GBSF1503241 | 2a | 2015 | Beef cattle | Gansu |
GYSF1503270 | 1a | 2015 | Yak | Gansu |
GBSF1503288 | 1a | 2015 | Beef cattle | Gansu |
GBSF1505314 | 2a | 2015 | Beef cattle | Gansu |
SBSF1505331 | 2a | 2015 | Beef cattle | Shanxi |
GBSF1506340 | Xv | 2015 | Beef cattle | Gansu |
GBSF1507358 | 1a | 2015 | Beef cattle | Gansu |
GBSF1509369 | 2a | 2015 | Beef cattle | Gansu |
GBSF1510375 | 2a | 2015 | Beef cattle | Gansu |
GBSF1510390 | 2a | 2015 | Beef cattle | Gansu |
QYSF1511395 | 2a | 2015 | Yak | Qinghai |
GBSF1511401 | 2a | 2015 | Beef cattle | Gansu |
GYSF1511409 | 2a | 2015 | Yak | Gansu |
SBSF1512413 | 4a | 2015 | Beef cattle | Shanxi |
GBSF1512419 | 2b | 2015 | Beef cattle | Gansu |
GBSF1512425 | 2a | 2015 | Beef cattle | Gansu |
GBSF1512433 | 2a | 2015 | Beef cattle | Gansu |
GBSF1601015 | Xv | 2016 | Beef cattle | Gansu |
GBSF1601024 | Xv | 2016 | Beef cattle | Gansu |
TYSF1601031 | 2b | 2016 | Yak | Tibet |
GBSF1601064 | 2a | 2016 | Beef cattle | Gansu |
GYSF1601073 | 6 | 2016 | Yak | Gansu |
GBSF1602082 | 2a | 2016 | Beef cattle | Gansu |
QYSF1602094 | 6 | 2016 | Yak | Qinghai |
GBSF1602098 | 2a | 2016 | Beef cattle | Gansu |
GBSF1602103 | 2a | 2016 | Beef cattle | Gansu |
SBSF1603115 | 4a | 2016 | Beef cattle | Shanxi |
SBSF1603121 | 4a | 2016 | Beef cattle | Shanxi |
GBSF1603138 | 2a | 2016 | Beef cattle | Gansu |
GBSF1603149 | 2a | 2016 | Beef cattle | Gansu |
QYSF1603158 | 6 | 2016 | Yak | Qinghai |
SBSF1604173 | 2a | 2016 | Beef cattle | Shanxi |
SBSF1604195 | 4a | 2016 | Beef cattle | Shanxi |
GBSF1605203 | Xv | 2016 | Beef cattle | Gansu |
GBSF1608241 | 2a | 2016 | Beef cattle | Gansu |
GYSF1610256 | 2a | 2016 | Yak | Gansu |
GYSF1610266 | 6 | 2016 | Yak | Gansu |
GBSF1610275 | 1a | 2016 | Beef cattle | Gansu |
GBSF1611283 | 1a | 2016 | Beef cattle | Gansu |
GBSF1611290 | 2a | 2016 | Beef cattle | Gansu |
Antimicrobial susceptibility testing
Antibiotic | Antimicrobial resistance rate No. (%) | |||||
---|---|---|---|---|---|---|
Total (n = 54) | Gansu (n = 37) | Shanxi (n = 8) | Xinjiang (n = 2) | Qinghai (n = 5) | Tibet (n = 2) | |
Penicillin G (P) | 54 (100%) | 37 (100%) | 8 (100%) | 2 (100%) | 5 (100%) | 2 (100%) |
Ampicillin (AMP) | 51 (94.44%) | 37 (100%) | 8 (100%) | 2 (100%) | 3 (60%) | 1 (50%) |
Amoxycillin/Clavulanic acid (AMC) | 5 (9.62%) | 3 (8.11%) | 1 (12.5%) | 1 (50%) | 0 | 0 |
Cephalothin (KF) | 29 (53.70%) | 19 (51.35%) | 5 (62.5%) | 2 (100%) | 2 (40%) | 1 (50%) |
Cephazolin (KZ) | 34 (62.96%) | 21 (56.76%) | 7 (87.5%) | 2 (100%) | 3 (60%) | 1 (50%) |
Cefamandole (MA) | 16 (29.63%) | 12 (32.43%) | 2 (25%) | 1 (50%) | 1 (20%) | 0 |
Cefoxitin (FOX) | 3 (5.56%) | 2 (5.41%) | 1 (12.5%) | 0 | 0 | 0 |
Ceftriaxone (CRO) | 12 (22.22%) | 9 (24.32%) | 2 (25%) | 1 (50%) | 0 | 0 |
Cefotaxime (CTX) | 14 (25.93%) | 10 (27.03%) | 2 (25%) | 1 (50%) | 1 (20%) | 0 |
Cefoperazone (CFP) | 6 (11.11%) | 6 (16.22%) | 0 | 0 | 0 | 0 |
Cefepime (FEP) | 0 | 0 | 0 | 0 | 0 | 0 |
Meropenem (MEM) | 0 | 0 | 0 | 0 | 0 | 0 |
Imipenem (IPM) | 0 | 0 | 0 | 0 | 0 | 0 |
Norfloxacin (NOR) | 16 (29.63%) | 12 (32.43%) | 3 (37.5%) | 1 (50%) | 0 | 0 |
Enrofloxacin (ENR) | 13(24.07%) | 11 (29.73%) | 2 (25%) | 0 | 0 | 0 |
Levofloxacin (LEV) | 14 (25.93%) | 11 (29.73%) | 2 (25%) | 1 (50%) | 0 | 0 |
Ciprofloxacin (CIP) | 2 (3.70%) | 2 (5.41%) | 0 | 0 | 0 | 0 |
Erythromycin (E) | 46 (85.19%) | 35 (94.59%) | 6 (75%) | 2 (100%) | 3 (60%) | 0 |
Tetracycline (TE) | 49 (90.74%) | 35 (94.59%) | 8 (100%) | 2 (100%) | 3 (60%) | 1 (50%) |
Chloramphenicol (C) | 17 (31.48%) | 10 (27.03%) | 6 (75%) | 1 (50%) | 0 | 0 |
Streptomycin (S) | 38 (70.37%) | 30 (81.08%) | 4 (50%) | 2 (100%) | 2 (40%) | 0 |
Gentamicin (CN) | 29 (53.70%) | 23 (62.16%) | 4 (50%) | 2 (100%) | 0 | 0 |
Amikacin (AK) | 3 (5.56%) | 3 (8.11%) | 0 | 0 | 0 | 0 |
Cephalosporin and/or Fluorquinolones resistance spectrum | Cephalosporin and/or Fluorquinolones resistance rate No. (%) | ||||
---|---|---|---|---|---|
Total (n = 26) | Gansu (n = 22a) | Shanxi (n = 2) | Qinghai (n = 1a) | Tibet (n = 1a) | |
KF/KZ | 5 (19.23%) | 3 (13.64%) | 0 | 1a (100%) | 1a (100%) |
KF/KZ/MA | 3 (11.54%) | 2a (9.09%) | 1 (50%) | 0 | 0 |
KF/KZ/MA/CRO | 1 (3.85%) | 1 (4.55%) | 0 | 0 | 0 |
KF/KZ/MA/CTX | 1 (3.85%) | 1 (4.55%) | 0 | 0 | 0 |
KF/KZ/MA/CRO/CFP | 1 (3.85%) | 1 (4.55%) | 0 | 0 | 0 |
KF/KZ/MA/FOX/CRO/CTX/CFP | 1 (3.85%) | 1 (4.55%) | 0 | 0 | 0 |
NOR/LEV | 3 (11.54%) | 3 (13.64%) | 0 | 0 | 0 |
ENR/LEV | 3 (11.54%) | 3 (13.64%) | 0 | 0 | 0 |
NOR/ENR/LEV/CIP | 1 (3.85%) | 1 (4.55%) | 0 | 0 | 0 |
KF/KZ/MA/CRO/CTX/NOR/LEV | 2 (7.69%) | 1 (4.55%) | 1 (50%) | 0 | 0 |
KF/KZ/MA/CRO/CTX/NOR/ENR/LEV | 1 (3.85%) | 1 (4.55%) | 0 | 0 | 0 |
KF/KZ/MA/CTX/CFP/CIP/NOR/ENR | 2 (7.69%) | 2 (9.09%) | 0 | 0 | 0 |
KF/KZ/MA/CRO/CTX/CFP/NOR/ENR/CIP | 1 (3.85%) | 1 (4.55%) | 0 | 0 | 0 |
KF/KZ/MA/FOX/CRO/CTX/CFP/NOR/ENR/LEV | 1 (3.85%) | 1 (4.55%) | 0 | 0 | 0 |
ARGs analysis of cephalosporin- and/or fluoroquinolone-resistant S. flexneri 2a isolates
Strain name | Antimicrobial spectrum | ARGs in plasmid | ||
---|---|---|---|---|
TEM
|
OXA
|
CTX-M-9
| ||
TYSF1412001 | KF/KZ |
TEM-1
|
OXA-1
| ---- |
QYSF1511395 | KF/KZ |
TEM-1
| ---- | ---- |
GBSF1503241 | KF/KZ |
TEM-1
|
OXA-1
|
CTX-M-14
|
GBSF1502176 | KF/KZ |
TEM-1
|
OXA-1
|
CTX-M-14
|
GBSF1602082 | KF/KZ |
TEM-1
| ---- |
CTX-M-14
|
SBSF1505331 | KF/KZ/MA |
TEM-1
|
OXA-1
|
CTX-M-14
|
GYSF1511409 | KF/KZ/MA |
TEM-1
| ---- | ---- |
GYSF1610256 | KF/KZ/MA |
TEM-1
|
OXA-1
| ---- |
GBSF1510375 | KF/KZ/MA/CRO |
TEM-1
|
OXA-1
|
CTX-M-14
|
GBSF1501105 | KF/KZ/MA/CTX |
TEM-1
|
OXA-1
|
CTX-M-14
|
GBSF1412056 | KF/KZ/MA/CRO/CFP |
TEM-1
|
OXA-1
|
CTX-M-14
|
GBSF1602103 | KF/KZ/MA/FOX/CRO/CTX/CFP |
TEM-1
|
OXA-1
|
CTX-M-14
|
GBSF1601064 | KF/KZ/MA/CRO/CTX/NOR/LEV |
TEM-1
| ---- |
CTX-M-14
|
SBSF1604173 | KF/KZ/MA/CRO/CTX/NOR/LEV |
TEM-1
|
OXA-1
|
CTX-M-14
|
GBSF1611290 | KF/KZ/MA/CTX/CFP/NOR/ENR |
TEM-1
|
OXA-1
|
CTX-M-14
|
GBSF1501026 | KF/KZ/MA/CTX/CFP/NOR/ENR |
TEM-1
|
OXA-1
|
CTX-M-14
|
GBSF1512425 | KF/KZ/MA/CRO/CTX/NOR/ENR/LEV |
TEM-1
|
OXA-1
| ---- |
GBSF1602098 | KF/KZ/MA/CRO/CTX/CFP/NOR/ENR/CIP |
TEM-1
|
OXA-1
|
CTX-M-14
|
GBSF1512433 | KF/KZ/MA/FOX/CRO/CTX/CFP/NOR/ENR/LEV |
TEM-1
|
OXA-1
|
CTX-M-14
|
Strain name | Antimicrobial spectrum | QRDR | ARGs in plasmid | ||||
---|---|---|---|---|---|---|---|
gyrA
|
parC
|
aac(6′)-Ib-cr
|
qnrS
|
qepA
| |||
83 | 87 | 80 | |||||
GBSF1509369 | NOR/LEV | S83 L | D87N | S80I | + | − | − |
GBSF1511401 | NOR/LEV | S83 L | D87N | S80I | + | − | − |
GBSF1608241 | NOR/LEV | S83 L | D87N | S80I | + | − | − |
GBSF1510390 | ENR/LEV | S83 L | D87D | S80I | + | + | − |
GBSF1603138 | ENR/LEV | S83 L | D87N | S80I | + | − | − |
GBSF1603149 | ENR/LEV | S83 L | D87N | S80I | + | − | − |
GBSF1505314 | NOR/ENR/LEV/CIP | S83 L | D87Y | S80I | + | + | − |
GBSF1601064 | KF/KZ/MA/CRO/CTX/NOR/LEV | S83 L | D87N | S80I | + | − | − |
SBSF1604173 | KF/KZ/MA/CRO/CTX/NOR/LEV | S83 L | D87N | S80I | + | − | − |
GBSF1611290 | KF/KZ/MA/CTX/CFP/NOR/ENR | S83 L | D87N | S80I | + | − | − |
GBSF1501026 | KF/KZ/MA/CTX/CFP/NOR/ENR | S83 L | D87N | S80I | + | + | − |
GBSF1512425 | KF/KZ/MA/CRO/CTX/NOR/ENR/LEV | S83 L | D87N | S80I | + | + | − |
GBSF1602098 | KF/KZ/MA/CRO/CTX/CFP/NOR/ENR/CIP | S83 L | D87Y | S80I | + | − | + |
GBSF1512433 | KF/KZ/MA/FOX/CRO/CTX/CFP/NOR/ENR/LEV | S83 L | D87N | S80I | + | + | − |