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Antimicrobial resistance and virulence genes in staphylococci isolated from aviary capercaillies and free-living birds in south-eastern Poland Cover

Antimicrobial resistance and virulence genes in staphylococci isolated from aviary capercaillies and free-living birds in south-eastern Poland

Open Access
|Sep 2022

Figures & Tables

Multidrug resistance profiles of isolated Staphylococcus strains

Species of birdNumber of Staphylococcus (% strains of all isolated strains)Phenotypic resistance combination pattern (number of classes of antimicrobial agents)Number of strains
no resistance10
β-lactams (1)6
macrolides (1)7
fusidic acid (1)6
fluoroquinolones (1)1
fusidic acid, fluoroquinolones (2)5
phenicols, tetracyclines (2)1
fluoroquinolones, β-lactams (2)1
fluoroquinolones, fusidic acid, β-lactams (3)2
fluoroquinolones, macrolides, β-lactams (3)2
macrolides, β-lactams, tetracyclines (3)2
fluoroquinolones, fusidic acid, tetracyclines (3)1
Capercaillie fluoroquinolones, fusidic acid, β-lactams, sulfonamide + dihydrofolate reductase inhibitor (4)1
(Tetrao urogallus)57 (44.2)fluoroquinolones, tetracyclines, β-lactams, sulfonamide + dihydrofolate reductase inhibitor (4)1
fluoroquinolones, fusidic acid, β-lactams, tetracyclines (4)1
fluoroquinolones, fusidic acid, β-lactams, macrolides (4)1
fluoroquinolones, tetracyclines, macrolides, sulfonamide + dihydrofolate reductase inhibitor (4)1
fluoroquinolones, macrolides, β-lactams, sulfonamide + dihydrofolate reductase inhibitor (4)1
fluoroquinolones, aminoglycosides, β-lactams, sulfonamide + dihydrofolate reductase inhibitor (4)1
fluoroquinolones, macrolides, β-lactams, tetracyclines, phenicols (5)1
fluoroquinolones, macrolides, β-lactams, tetracyclines, sulfonamide + dihydrofolate reductase inhibitor (5)2
fluoroquinolones, macrolides, β-lactams, Fusidic acid, aminoglycosides, sulfonamide + dihydrofolate reductase inhibitor (6)1
fluoroquinolones, macrolides, β-lactams, Fusidic acid, aminoglycosides, tetracyclines, phenicols, sulfonamide + dihydrofolate reductase inhibitor (8)2
no resistance7
β-lactams (1)1
macrolides (1)1
tetracyclines (1)2
sulfonamide + dihydrofolate reductase inhibitor (1)1
macrolides, tetracyclines (2)2
Fusidic acid, tetracyclines (2)3
Fusidic acid, fluoroquinolones (2)1
macrolides, β-lactams (2)1
White stork (Ciconia ciconia)27 (20.9)fusidic acid, fluoroquinolones, tetracyclines (3) fusidic acid, fluoroquinolones, β-lactams (3)1 1
fusidic acid, fluoroquinolones, β-lactams, macrolides (4)1
fusidic acid, fluoroquinolones, macrolides, sulfonamide + dihydrofolate reductase inhibitor (4)1
fusidic acid, fluoroquinolones, macrolides, β-lactams, tetracyclines (5)1
fusidic acid, fluoroquinolones, macrolides, tetracyclines, sulfonamide + dihydrofolate reductase inhibitor (5)1
fusidic acid, fluoroquinolones, macrolides, β-lactams, tetracyclines, phenicols (6)1
fusidic acid, fluoroquinolones, aminoglycosides, β-lactams, tetracyclines, phenicols (6)1
no resistance1
phenicols (1)1
β-lactams, tetracyclines (2)1
fluoroquinolones, macrolides, tetracyclines (3)4
macrolides, β-lactams, tetracyclines (3)1
fluoroquinolones, β-lactams, tetracyclines (3)1
fluoroquinolones, macrolides, fusidic acid, tetracyclines (4)2
fluoroquinolones, β-lactams, fusidic acid, tetracyclines (4)4
Common pheasant fluoroquinolones, β-lactams, tetracyclines, sulfonamide + dihydrofolate reductase inhibitor (4)1
(Phasianus colchicus)22 (17.1)fluoroquinolones, fusidic acid, tetracyclines, sulfonamide + dihydrofolate reductase inhibitor (4)1
fluoroquinolones, β-lactams, tetracyclines, macrolides (4)1
fluoroquinolones, fusidic acid, tetracyclines, macrolides, phenicols (5)1
fluoroquinolones, fusidic acid, β-lactams, tetracyclines, sulfonamide + dihydrofolate reductase inhibitor (5)1
fluoroquinolones, fusidic acid, β-lactams, tetracyclines, macrolides, sulfonamide + dihydrofolate reductase inhibitor (6)1
fluoroquinolones, fusidic acid, β-lactams, tetracyclines, macrolides, phenicols, sulfonamide + dihydrofolate reductase inhibitor (7)1
Tawny owl no resistance4
6 (4.6)β-lactams (1)1
(Strix aluco) macrolides, fusidic acid, tetracyclines (3)1
Common kestrel5 (3.9)no resistance4
(Falco tinnunculus) fluoroquinolones, fusidic acid, macrolides (3)1
Common blackbird (Turdus merula)4 (3.1)no resistance4
no resistance1
Mute swan3 (2.3)fluoroquinolones, β-lactams (2)1
(Cygnus olor) fluoroquinolones, β-lactams, fusidic acid (3)1
Song thrush no resistance1
(Turdus3 (2.3)fluoroquinolones, fusidic acid (2)1
philomelos) β-lactams, macrolides (2)1
Grey heron (Ardea cinerea)1 (0.8)macrolides, aminoglycosides (2)1
Great spotted woodpecker (Dendrocopos major)1 (0.8)no resistance1

Nucleotide sequences and sizes of PCR products of amplified genes

GeneOligonucleotide sequence (5′-3′)Amplicon size (bp)PCR conditionsReference
mecAAAAATCGATGGTAAAGGTTGGC AGTTCTGGCACTACCGGATTTGC53394℃, 5 min, 40 cycles of 94℃ for 1 min, 58℃ for 1 min, 72℃ for 2 min, final extension 72℃ for 5 min(23)
mecCGAA AAA AAG GCT TAG AAC GCC TC GAA GAT CTT TTC CGT TTT CAG C13894℃, 15 min, 30 cycles of 94℃ for 30 s, 59℃ for 1 min, 72℃ for 1 min, final extension for 10 min(35)
blaZ ermAACTTCAACACCTGCTGCTTTC TAGGTTCAGATTGGCCCTTAG TCT AAAAAG CATGTAAAAGAA TGA TTATAATTATTT GATAGC TTC173 64595℃, 3 min, 30 cycles of 95℃ for 30 s, 54℃ for 30 s, 72℃ for 30 s, final extension 72℃ for 4 min(7)
ermBTAACGACGAAACTGGCTAAAA ATCTGTGGTATGGCGGGTAAG41695℃, 3 min, 30 cycles of 95℃ for 30 s, 55℃ for 30 s, 72℃ for 45 s, final extension 72℃ for 5 min(36)
ermCTAATCGTGGAATACGGGTTTG AATCGTCAATTCCTGCATGT299
tetKGTAGCGACAATAGGTAATAGT GTAGTGACAATAAACCTCCTA360
tetMCATATGTCCTGGCGTGTCTA AGTGGAGCGATTACAGAA15894℃, 5 min, 30 cycles of 94℃ for 1 min, 57℃ for 1 min, 72℃ for 1 min,(18)
tetLATAAATTGTTTCGGGTCGGTAAT AACCAGCCAACTAATGACAATGAT1077final extension 72℃ for 5 min
tetOAACTTAGGCATTCTGGCTCAC TCCCACTGTTCCATATCGTCA514
msrA/BGCAAATGGTGTAGGTAAGACAACT ATCATGTGATGTAAACAAAAT39995℃, 3 min, 35 cycles of 93℃ for 30 s, 55℃ for 2 min, 74℃ for 1 min, final extension 72℃ 90 s(36)
Aac(6′)/aph(2ʺ)CAGAGCCTTGGGAAGATGAAG CCTCGTGTAATTCATGTTCTGGC34894℃, 10 min, 35 cycles of 94℃ for 45 s, 60℃ for 60 s, 72℃ for 60 s, final extension cycle 72℃ for 5 min(39)
norATTTGTTTTCAGTGTCAGAATTTATGTTTG GGCTTGGTGAAATATCAGCTATTAAAC14094℃, 5 min, 30 cycles of 94℃ for 30 s, 60℃ for 30 s, 72℃ for 60 s, final extension 72℃ for 5 min(26)
cfrTGA AGT ATA AAG CAG GTT GGG AGT CA ACC ATA TAA TTG ACC ACA AGC AGC74694℃, 2 min, 30 cycles of 94℃ for 30 s, 45℃ for 30 s, and 72℃ for 45 s, final extension 72℃ for 1 min(13)
mphCGAG ACT ACC AAG AAG ACC TGA CG CAT ACG CCG ATT CTC CTG AT53095℃, 5 min, 35 cycles of 94℃ for 1 min, 59℃ for 1 min, 72℃ for 2 min, final extension 72℃ for 1 min(16)
seaTGCATGTTTTCAGAGTTAATC ACGATCAATTTTTACAGC544
sebTCTTTGTCGTAAGATAAACTTC GAATGATATTAATTCGCATC41694℃, 5 min, 35 cycles of 94℃ for 2 min, 57℃ for 2 min, 72℃ for 1 min,(22)
secGACATAAAAGCTAGGAATTT AAATCGGATTAACATTATCCA257final extension 72℃ for 7 min
sedTTACTAGTTTGGTAATATCTCCTT CCACCATAACAATTAATGC334
seeATAGATAAAGTTAAAACAAGCAA TAACTTACCGTGGACCC170
tstACCCCTGTTCCCTTATCATC TTTTCAGTATTTGTAACGCC326
pvlATCATTAGGTAAAATGTCTGGACATGATCCA GCATCAASTGTATTGGATAGCAAAAGC43395℃, 5 min, 30 cycles of 94℃ for 1 min, 55℃ for 30 s, 72℃ for 1 min,(38)
etaGCAGGTGTTGATTTAGCATT AGATGTCCCTATTTTTGCTG93final extension at 72℃ for 5 min
etbACAAGCAAAAGAATACAGCG GTTTTTGGCTGCTTCTCTTG226

Total number of birds and number and type of organs from which bacteria of the genus Staphylococcus were isolated

Number of Staphylococcus strains isolated from individual organs or tissue
Bird speciesTotal examined animalsHeartLiverSpleenTarsal jointConjunctivaPalatal fissureYolk sacDead embryo
Capercaillie (Tetrao urogallus)73559457913
White stork (Ciconia ciconia)35335376--
Common pheasant(Phasianus colchicus)461361542-
Tawny owl (Strix aluco)10-12-12--
Common kestrel (Falco tinnunculus)81-2--2--
Common blackbird (Turdus merula)7---112--
Mute swan (Cygnus olor)5-1--11--
Song thrush (Turdus philomenos)5--1--2--
Grey heron (Ardea cinerea)2-1------
Great spotted woodpecker (Dendrocopos major)4--1-----
Common buzzard (Buteo buteo)3--------
Black grouse (Lyrurus tetrix)2--------
Common swift (Apus apus)4--------
Meadow pipit (Anthus pratensis)5--------
Fieldfare (Turdus pilaris)4--------
Lesser spotted eagle (Clanga pomarina)1--------
Total214101426920261113

Percentage of Staphylococcus strains isolated from individual bird species resistant to specific antibacterial agents

Bird speciesNumber of strainsAntibiotic
AML25AMC30AMP10P10FOX30DA2C30E15CN10TE30SXT25ENR5FD5
Capercaillie (Tetrao urogallus)5712 21%7 12.3%21 36.8%18 31.6%9 15.8%23 40.4%4 7%19 33.3%4 7%14 24.6%11 19.3%26 45.6%20 35.1%
White stork (Ciconia ciconia)275 18.5%3 11.1%6 22.2%5 18.5%5 18.5%3 11.1%2 7.4%9 33.3%1 3.7%12 44.4%3 11.1%9 33.3%12 44.4%
Common pheasant (Phasianus colchicus)2210 45.5%8 36.4%12 54.5%11 50.0%7 31.8%3 13.6%3 13.6%11 50.5%-20 90/9%5 22.7%18 81.8%11 50.5%
Tawny owl (Strix aluco)6--1 16.7%----1 16.7%-1 16.7%--1 16.7%
Common kestrel (Falco tinnunculus)5-------20.01 %---20.01 %20.01 %
Common blackbird (Turdus merula)4-------------
Mute swan (Cygnus olor)31 33.3%1 33.3%2 66.6%2 66.6%2 66.6%------2 66.6%1 33.3%
Song thrush (Turdus philomenos)3-1 33.3%1 33.3%1 33.3%---1 33.3%---1 33.3%1 33.3%
Grey heron (Ardea cinerea)1-------1 100%1 100%----
Great spotted woodpecker (Dendrocopus major)1-------------

Presence of genes encoding resistance to antimicrobial agents and staphylococcal toxins in isolated strains

Staphylococcus species
GeneS. sci (n=56)S. xyl (n=14)S. equ (n=12)S. sap (n=8)S. aur (n=6)S. epi (n=6)S. coh (n=6)S. pse (n=3)S. hae (n=3)S. len (n=3)S. klo (n=2)S. suc (n=2)S. vit (n=2)S. con (n=1)S. nep (n=1)S. arl (n=1)S. pas (n=1)S. chr (n=1)S. war (n=1)Total%
mecA7 1 1 97.0
mecC
blaZ23 242 1 11 1612.4
msrA/B3233 1121 1 1713.2
ermA5 1 64.7
ermB12121 22 1 2116.3
ermC7351 5311 1 2720.9
tetK195452442 21 1 4938.0
tetM1675331 1 1 1 11 4031.0
tetL5 1 1 75.4
tetO311 1 1 75.4
cfr 1 1 1 1 1 53.9
norA62116211 2015.5
aac(6′)/ aph(2ʺ) 31 1 11 75.4
mphC75 3222 1 1 2317.8
sea 1 1 1 32.3
seb
sec
sed
see
pvl
eta
etb
tst1 10.8
129

Phenotypic antimicrobial resistance of Staphylococcus strains

Antibiotic
Species AML25AMC30AMP10P10FOX30DA2C30E15CN10TE30SXT25ENR5FD5
S. sciuri, n = 56R15818161422-11532628
I-----12-511---
S. xylosus, n = 14R32331-24-6266
I-----112-----
S. equorum, n = 12R1-111122-3112
I-------31----
S. saprophyticus, n = 8R-121---5-7251
I-------------
S. aureus, n = 6R1444-------31
I-----3-4-----
S. epidermidis, n = 6R1143-3-4-455-
I-------------
S. cohnii, n = 6R313232-3-3232
I-----1-21----
S. pseudintermedius, n = 3R1111-111-1-1-
I-------1-----
S. haemolyticus, n = 3R1-2211-111211
I-------------
S. lentus, n = 3R---------1-11
I-------------
S. kloosii, n = 2R--1------1--1
I-------1-----
S. succinus, n = 2R-------1-----
I-------------
S. vitulinus, n = 2R-1111--1---11
I-------------
S. condimenti, n =1R1-11111----11
I-------1111--
S. nepalensis, n = 1R---------1-1-
I------1------
S. arlettae, n = 1R---1---1-1- 11
I-------------
S. pasteuri, n = 1R--1----------
I-------------
S. chromogenes, n = 1R111111-1-1111
I-------------
S. warneri, n = 1R-------------
I-------------
Total number 282043372329943647195747
% 21.715.533.328.717.822.57.033.34.736.414.744.236.4
Language: English
Page range: 361 - 372
Submitted on: Mar 23, 2022
Accepted on: Sep 7, 2022
Published on: Sep 15, 2022
Published by: National Veterinary Research Institute in Pulawy
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year

© 2022 Magdalena Sulikowska, Agnieszka Marek, Łukasz Jarosz, Dagmara Stępień-Pyśniak, Renata Urban-Chmiel, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.