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Phenotypic and genetic properties of susceptible and multidrug-resistant Pseudomonas aeruginosa isolates in Southern Serbia Cover

Phenotypic and genetic properties of susceptible and multidrug-resistant Pseudomonas aeruginosa isolates in Southern Serbia

Archives of Industrial Hygiene and Toxicology
Volume 71 (2020): Issue 3 (September 2020)
By: ,  ,  ,  ,  ,  ,   and    
Open Access
|Oct 2020
Figures & tables

Figures & Tables

Table 1

Serological identification of polyvalent and monovalent serotypes, pigmentation, and biofilm formation of Pseudomonas spp. clinical isolate of different origin

Polyvalent and monovalent serotypesPigments typeBiofilm formation
Isolate originPMAPMEPMCPMFNTPyov+ PyocPyovPyocCat. 0(-)Cat. 1(+)Cat. 2(++)Cat. 3(+++)
P1P6P5P9P10P11
Throat44--113931265-
Tongue-1--1--11-1-1-
Wound7736-813301221217141
Cer-vagmal canal------211-1-1-
Ear-2---11211--4-
Sputum14-1-127112412
Urine culture43-2-47162231232
Σtotal isloates16213921528662172139295

[i] NT – non-typeable; Pyov – pyoverdine; Pyoc – pyocyanin; Cat. 0 (-) – not a biofilm producer; Cat. 1 (+) – weak biofilm producer; Cat. 2 (++) – moderate biofilm producer; Cat. 3 (+++) – strong biofilm producer

Table 2

Antibiotic susceptibility of inpatient and outpatient Pseudomonas spp. isolates

Inpatients
Isolate coded nameSpecimenAminoglycosidesFluoroquinolonesPolypMonobPenicillin comb.CarbapenemsCephalosporins
AmikGentNetTobrOfloxCiprLevoflColAztrPip/tazDorImpMerCeftCef
5660ThroatSSSSSSSSSSSSSSS
5661ThroatSSSSSSSSSSSSSSS
1183UrineSRRRRRRSSSSSRRS
2638UrineRRRRRRRSSRRRRRR
2844UrineSRSRRRRSSSSSSSR
3853UrineSRRRRRRSSSSSSRS
611WoundSSSSSSSSSSSSSSS
821WoundSSSSSSSSSSSSRSS
971WoundSSSSSSSSSSSSSRR
1087WoundRRRRRRRSSRRRRRR
1416WoundSSSSRSSSSSSSSSS
2064WoundSSRSSSSSSSSSSRS
2073WoundRRRRRRRSSSRRRRR
2581WoundRRRRRRRSSRSSSRS
2645WoundRRRRRRRSSRRRRRR
2900WoundRRRRRRRSSSRRRRR
3122WoundSSRSRSSSSSSSSSS
3322WoundRRRRRRRSSRSSRSS
3451WoundSRRRRRRSSRSSSRS
3563WoundRRRRRRRSSSSSSSS
3595WoundSSSSSSSSSSSSSSS
3658WoundSRRRRRRSSSRSSSS
3714WoundSSSRSSSSRSSRRRR
3883WoundSSSSSSSSSSSSSSS
4071WoundSSSSSSSSSSSSSSS
4082WoundSSSSSSSSSSSSSSS
4211WoundSSSSSSSSSSSSSRS
4212WoundSSSSSSSSSSSSSSS
4312WoundRRRSRRSSSSSSSRS
4314WoundSSSSSSSSSSSSSSS
4354WoundSSSSSSSSSSSSSSS
4473WoundSSSSSSSSSSSSSSS
4541WoundSSSSSSSSSSSSSSS
5348WoundRRRRRRRSSRSSSSS
5774WoundRRRRRRRSSSSSISR
5797WoundRRRRRRRSSSSSSRS
6412WoundSSSSSSSSSSSSSSS
6982WoundSSSSSSSSSSSSSSS
7543WoundRRRRRRRSSRSSSRR
7698WoundRRIRSISSSSSSSSS
7881WoundSSRSSSSSSSSSSSS
8283WoundIIRRRRRSSSSSSSS
8702WoundSSSSSSSSSSSSSSS
8913WoundRRRRRRRSSSSSSSS
10195WoundSSSSRIISSSSSSRR
10336WoundSSSSSSSSSSSSSSS
10708WoundRIISSSSSSSSSSSS
10800WoundSISSSSSSSSRRSRS
11947WoundSSSSSSSSSSSSSSS
19677WoundSSSSSSSSSSSSSSS
Outpatients
Isolate coded nameSpecimenAminoglycosidesFluoroquinolonesPolypMonobPenicillin comb.CarbapenemsCephalosporins
AmikGentNetTobrOfloxCiprLevoflColAztrPip/tazDorImipMerCeftCef
2609EarSSSSSSSSSRSSSSS
4278EarSISSSSSSSSSSSSS
5518EarSSSSSSSSSSSSRRR
7546EarSSSSSSSSSSSSSSS
294SputumSRSISSSSSSSSSSS
2124SputumSSSRRRRSSRSSSRR
2941SputumSSSSSSSSSSSSSSS
2966SputumSSSSSSSSSSSSSSS
3496SputumSSSSSSSSSSSSSRR
3919SputumSSSSSSSSSSSSSSS
8142SputumSRSSSSSSSSSSSSS
9921SputumRRRRRRRSSSSSSRR
11838SputumSRSSRSSSSSSSSSS
1863ThroatSSSSSSSSSSSSSSS
2005ThroatSSSSSSSSSSSSSSS
2047ThroatSSSSSSSSSSSSSSS
3238ThroatSSSSSSSSSSSSSSS
3477ThroatSSSSSSSSSSSSSSS
3540ThroatSSSSSSSSSSSSSSS
3864ThroatSISSSSSSSSSSSSS
4087ThroatSSSSSSSSSSSSSSS
4646ThroatSSSSSSSSSSSSSSS
9642ThroatSSSSSSSSSSSSSSS
10412ThroatSSSSSSSSSSSSSSS
2383TongueSRSSSSSSSRSSSSS
2967TongueIRRRSSSSSRSSSRR
1408UrineRRSRRRRSSSSSSSS
1790UrineIRSSSSSSSSSSSSS
2285UrineSSSSSSSSSSSSSSS
2588UrineRRRRRRISSSSSRSS
2597UrineSRRRRRRSRSSSSSR
2711UrineRIRRSSRSSSSRSSR
2875UrineRRSRRRRSSSSSSSS
3199UrineSRRRRRRSSSSSSSS
3214UrineSRRRRRRSSRSSSRR
3777UrineSRRRRRRSSSRSRSR
4188UrineIRRRRRRSSSRSRSR
4362UrineSSSSSSSSSSSSSRR
5586UrineSSSSSSSSSSSSSSS
8599UrineSRRRRRRSSSSSRRS
10019UrineRRRRRRRSSISSSSR
10600UrineSSSSSSSSSRSSSRS
2671Vag. SwabSSSSSSSSSSSSSSS
2689Vag. SwabSSSSSSSSSSSSSRS

[i] Amik – amikacin; Gent – gentamicin; Net – netilmicin; Tobr – tobramycin; Oflox – ofloxacin; Cipr – ciprofloxacin; Levofl – levofloxacin; Col – colistin; Aztr – aztreonam; Pip/taz – piperacillin/ tazobactam; Dor – doripenem; Imip – imipenem; Mer – meropenem; Ceft – ceftazidime; Cef – cefepime. Polyp – polypeptide class; Monob – monobactams class. S – susceptible; I – intermediate susceptible; R– resistant isolates. Bolded letters represent resistant values

Figure 1

Susceptibility of clinical isolates of Pseudomonas spp. on different antibiotic classes. The isolates were classified as resistant (R), intermediate (I), or susceptible (S)

Table 3

Minimum inhibitory concentration (MIC, expressed in μg/mL) of inpatient and outpatient Pseudomonas spp. isolates and their potential to produce beta-lactamases

InpatientsOutpatients
Isolate coded nameSpecimenMerCeftCefEnzyme producedIsolate name codedSpecimenMerCeftCefEnzyme produced
5660Throat<148-2609Ear<1<2<2-
5661Throat<1<2<2-4278Ear<1<2<2-
1183Urine>16164-5518Ear>161632MBL
2638Urine>16>32>32CRP7546Ear<1<2<2-
2844Urine<1<216-294Sputum<1<2<2-
3853Urine4164-2124Sputum2>32>32-
611Wound<1<2<2-2941Sputum<1<2<2-
821Wound>1642-2966Sputum<124-
971Wound<13216-3496Sputum<1>32>32-
1087Wound>16>32>32CRP3919Sputum<144-
1416Wound<144-8142Sputum<144-
2064Wound<116<2-9921Sputum21616-
2073Wound>16>32>32CRP11838Sputum<1<2<2-
2581Wound<1168MBL1863Throat<1<2<2-
2645Wound>16>32>32CRP2005Throat<122-
2900Wound>16>32>32CRP2047Throat<1<2<2-
3122Wound<188-3238Throat<144-
3322Wound1622MBL3477Throat<122-
3451Wound<1168-3540Throat<142-
3563Wound<144-3864Throat<122-
3595Wound<144-4087Throat<144-
3658Wound<142-4646Throat<122-
3714Wound>1632>32MBL9642Throat<1<2<2-
3883Wound2<2<2-10412Throat<144-
4071Wound<144-2383Tongue<144-
4082Wound<144-2967Tongue<11632-
4211Wound<1168-1408Urine<1<2<2-
4212Wound<144-1790Urine<144-
4312Wound<1162-2285Urine<122-
4314Wound<122-2588Urine1624-
4354Wound<122-2597Urine<1416ESBL
4473Wound<188-2711Urine<1416MBL
4541Wound<1<2<2-2875Urine<122-
5348Wound<124-3199Urine<144-
5774Wound4416-3214Urine1>3216-
5797Wound2>322-3777Urine>16416CRP
6412Wound<122-4188Urine>16432CRP
6982Wound<1<2<2-4362Urine<11616-
7543Wound11616ESBL5586Urine<1<2<2-
7698Wound<144-8599Urine>16328-
7881Wound<122-10019Urine<1816-
8283Wound<1<2<2-10600Urine<1164-
8702Wound<1<2<2-2671Vag. swab<122-
8913Wound<1<2<2-2689Vag. swab<1168-
10195Wound<1>32>32-
10336Wound<1<2<2-
10708Wound<142-
10800Wound<1328CRP
11947Wound<122-
19677Wound<144-

[i] Mer – meropenem (S<2, I4-8, R>8); Ceft – ceftazidime (S<8; R>8); Cef – cefepime (S<8; R>8); CRP – carbapenemase-producnig Pseudomonas sp.; MBL – metallo-beta-lactamase; ESBL – extended spectrum beta-lactamase. Bolded numbers represent resistant MIC values. The results are expressed in μg/mL

Table 4

Multidrug resistant Pseudomonas spp. isolates and their pigmentation, serotype, and biofilm formation potential

graphic/j_aiht-2020-71-3418_ingr_001.png

[i] Pyov – Pyoverdine; Pyoc – Pyocyanin; NT – non-tippable; No biofilm producer (category -); Weak biofilm producer (category +); Moderate biofilm producer (category ++); Strong biofilm producer (category +++); Out – outpatient; In – inpatient; graphic/j_aiht-2020-71-3418_ingr_002.png Resistant up to four antibiotic classes or more; graphic/j_aiht-2020-71-3418_ingr_003.png Resistant up to three antibiotic classes; graphic/j_aiht-2020-71-3418_ingr_004.png Resistant up to two antibiotic classes; graphic/j_aiht-2020-71-3418_ingr_005.png Resistant up to one antibiotic class; graphic/j_aiht-2020-71-3418_ingr_006.png Susceptible to all tested antibiotic classes. Framed isolates have been selected for molecular genetic characterization

Table 5

Selected isolates for further molecular genetic analysis

Group IGroup IIGroup IIIGroup IVGroup V
Resistant up to 4 antibiotic classes or moreResistant up to 3 antibiotic classesResistant up to 2 antibiotic classesResistant up to 1 antibiotic classSusceptible to all tested antibiotic classes
Isolate coded nameGroup cipherIsolate coded nameGroup cipherIsolate coded nameGroup cipherIsolate coded nameGroup cipherIsolate coded nameGroup cipher
27111-157972-182833-126094-138645-1
75431-243122-255183-28214-220475-2
25971-353482-3101953-343624-364125-3
37141-4100192-435633-417904-432385-4
26451-536582-531223-578814-5119475-5
37771-629672-6108003-642114-6
29001-728442-720643-734964-7
34511-838532-8118383-881424-8
21241-925882-914083-99714-9
41881-1099212-1023833-1026894-10
Figure 2

Dendrogram obtained by UPGMA analysis of the selected Pseudomonas isolates using 208 (top) and 272 (bottom) primers, based on RADP profile (right). Genetic distances were presented with numbers placed on the branches. Rectangles are representing the isolate grouping according to obtained RAPD patterns

Figure 3

Dendrogram obtained by UPGMA analysis of the selected Pseudomonas isolates using BOX (top) and ERIC (bottom) primers, based on rep-PCR profile (right). Genetic distances were presented with numbers placed on the branches. Rectangles are representing the isolate grouping according to obtained rep-PCR patterns

Figure 4

Phylogenetic relationships of Pseudomonas aeruginosa clinical isolates from all 12 RAPD272 clusters based on the sequence of 16S rRNA. Phylogenetic tree was constructed by the neighbour-joining method and the distances were calculated with the Kimura two-parameter model. Bootstrap values are given for each node, with 1000 replicates. Stenotrophomonas maltophilia was used as an outgroup. The horizontal bar indicates a genetic distance of 0.02

DOI: https://doi.org/10.2478/aiht-2020-71-3418 | Journal eISSN: 1848-6312 (formerly 0004-1254) | Journal ISSN: 0004-1254
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Language: English, Croatian
Page range: 231 - 250
Submitted on: Apr 1, 2020
Accepted on: Aug 1, 2020
Published on: Oct 6, 2020
Published by: Institute for Medical Research and Occupational Health
In partnership with: Paradigm Publishing Services
Keywords:
clinical isolates,
multidrug resistance,
phenotypic characteristics,
RAPD PCR profiling
Related subjects:
Medicine,
Basic medical science,
Basic medical science, other

© 2020 Marko Milojković, Željka Nenadović, Slaviša Stanković, Dragana D. Božić, Nataša Stanković Nedeljković, Ivana Ćirković, Marija Petrović, Ivica Dimkić, published by Institute for Medical Research and Occupational Health
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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