High Prevalence of Antibiotic-Resistant Escherichia coli Isolates from Retail Poultry Products in Spain
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Sampling
2.2. Enterobacteriaceae Counts: Isolation and Identification of Escherichia coli
2.3. Phenotypic Sensitivity Test of E. coli Isolates
Growth Curves of Beta-Lactam- and Tetracycline-Resistant E. coli
2.4. Multiplex Polymerase Chain Reaction (PCR) for the Detection of Antibiotic Resistance Genes
2.4.1. Beta-Lactam Resistance Genes
2.4.2. Tetracycline Resistance Genes
2.5. Latex Agglutination Test for E. coli O157:H7
2.6. Statistical Analysis
3. Results
3.1. Enterobacteriaceae Counts: Isolation and Identification of Escherichia coli
3.2. Phenotypic Sensitivity Test of E. coli Isolates
Growth Curves of Beta-Lactam- and Tetracycline-Resistant E. coli
3.3. Multiplex PCR for Detection of Beta-Lactam and Tetracycline Resistance Genes
3.4. Latex Agglutination Test for E. coli O157:H7
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Antibiotic Group | Gene | Oligonucleotides | Band Size (pb) | Reference |
---|---|---|---|---|
Beta-lactam | blaTEM | 5′-TTAACTGGCGAACTACTTAC-3′ | 247 | [16] |
5′-GTCTATTTCGTTCATCCATA-3′ | ||||
blaSHV | 5′-AGGATTGACTGCCTTTTTG-3′ | 393 | [17] | |
5′-ATTTGCTGATTTCGCTCG-3′ | ||||
blaCMY-2 | 5′-GACAGCCTCTTTCTCCACA-3′ | 1000 | [16] | |
5′-TGGACACGAAGGCTACGTA-3′ | ||||
Tetracycline | tet(A) | 5′-GGCGGTCTTCTTCATCATGC-3′ | 502 | [18] |
5′-CGGCAGGCAGAGCAAGTAGA-3′ | ||||
tet(B) | 5′-CGCCCAGTGCTGTTGTTGTC-3′ | 173 | [19] | |
5′-CGCGTTGAGAAGCTGAGGTG-3′ |
Poultry Product | Retail | Enterobacteriaceae Log[CFU/g] | E. coli Presence | E. coli Log[CFU/g] (Proportion) |
---|---|---|---|---|
Hindquarters | Supermarket | 3.5 ± 0.1 c | + | 0.3 ± 0.0 (8%) |
5.2 ± 0.0 l | + | 1.5 ± 0.1 (30%) | ||
5.5 ± 0.0 m | + | 0.1 ± 0.0 (2%) | ||
6.5 ± 0.0 ñ | + | 0.2 ± 0.0 (3%) | ||
3.3 ± 0.1 a,b | + | 0.2 ± 0.0 (5%) | ||
5.2 ± 0.1 l | + | 0.8 ± 0.1 (16%) | ||
6.5 ± 0.0 ñ | - | - | ||
4.5 ± 0.1 h,i | + | 0.1 ± 0.0 (2%) | ||
4.5 ± 0.0 h,i | + | 0.1 ± 0.0 (3%) | ||
4.4 ± 0.0 g,h | + | 0.2 ± 0.0 (5%) | ||
4.0 ± 0.0 e | + | 0.1 ± 0.0 (3%) | ||
3.4 ± 0.0 b,c | + | 2.5 ± 0.1 (74%) | ||
3.2 ± 0.0 a | + | 0.5 ± 0.0 (15%) | ||
3.5 ± 0.0 c | + | 0.1 ± 0.0 (3%) | ||
4.5 ± 0.0 h,i | + | 0.7 ± 0.0 (16%) | ||
6.2 ± 0.1 n | - | - | ||
5.0 ± 0.0 k | + | 0.1 ± 0.0 (1%) | ||
4.6 ± 0.1 i | + | 0.2 ± 0.0 (5%) | ||
Butcher | 4.3 ± 0.0 g | + | 1.4 ± 0.2 (24%) | |
4.7 ± 0.1 j | - | - | ||
4.2 ± 0.1 f | + | 0.1 ± 0.0 (2%) | ||
3.4 ± 0.0 b,c | + | 0.1 ± 0.0 (4%) | ||
3.4 ± 0.0 b,c | + | 0.2 ± 0.0 (5%) | ||
5.0 ± 0.0 k | - | - | ||
Liver | Supermarket | 3.7 ± 0.1 d | + | 0.2 ± 0.0 (4%) |
5.4 ± 0.0 m | + | 0.5 ± 0.1 (9%) | ||
4.0 ± 0.0 e | + | 0.8 ± 0.1 (20%) | ||
4.5 ± 0.1 h,i | + | 0.2 ± 0.0 (5%) | ||
Butcher | 4.0 ± 0.1 e | + | 2.3 ± 0.2 (57%) | |
5.2 ± 0.1 l | + | 0.2 ± 0.0 (3%) |
Supermarket | Butcher | |||
---|---|---|---|---|
Resistance Phenotype | Hindquarters | Livers | Hindquarters | Livers |
Suscepetible | 0 | 0 | 1 | 0 |
Resistance to 1 | 2 | 0 | 1 | 0 |
Resistance to 2 | 5 | 6 | 3 | 0 |
Resistance to 3 | 18 | 4 | 27 | 0 |
Resistance to 4 | 20 | 5 | 11 | 0 |
Resistance to 5 | 29 | 3 | 6 | 0 |
Resistance to 6 | 38 | 2 | 1 | 4 |
Resistance to 7 | 22 | 1 | 0 | 5 |
Resistance to 8 | 12 | 1 | 0 | 5 |
Resistance to 9 | 6 | 0 | 0 | 2 |
Resistance to 10 | 0 | 0 | 0 | 1 |
Total | 152 | 22 | 50 | 17 |
Antibiotic | E. coli Isolate Code | Rate (h−1) | Generation Time (h) | ODmax | Latency Phase (h) |
---|---|---|---|---|---|
Ampicillin | 3.10 (high resistance) | 0.19 c ± 0.00 | 1.61 a ± 0.01 | 1.274 c ± 0.42 | 1.90 b ± 0.02 |
6.8 (medium resistance) | 0.18 b ± 0.00 | 1.71 b ± 0.04 | 0.891 b ± 0.01 | 1.79 a ± 0.18 | |
7.1 (sensitive) | 0.00 a ± 0.00 | >24 c | 0.084 a ± 0.00 | >24 c | |
7.15 (medium resistance) | 0.17 b ± 0.00 | 1.75 b ± 0.02 | 0.859 b ± 0.04 | 1.77 a ± 0.00 | |
19.27 (high resistance) | 0.17 b ± 0.00 | 1.75 b ± 0.03 | 1.099 c ± 0.05 | 2.02 b ± 0.13 | |
Amoxicillin | 3.10 (high resistance) | 0.18 b ± 0.01 | 1.69 a ± 0.06 | 1.010 c ± 0.03 | 1.94 c ± 0.01 |
6.8 (medium resistance) | 0.17 b ± 0.01 | 1.75 a ± 0.13 | 0.846 b ± 0.06 | 1.89 b ± 0.10 | |
7.1 (sensitive) | 0.00 a ± 0.00 | >24 c | 0.082 a ± 0.00 | >24 d | |
7.15 (medium resistance) | 0.16 b ± 0.00 | 1.86 b ± 0.02 | 0.904 b ± 0.13 | 1.86 a,b ± 0.04 | |
19.27 (high resistance) | 0.16 b ± 0.00 | 1.88 b ± 0.03 | 0.972 c ± 0.03 | 2.06 c ± 0.01 | |
Tetracycline | 3.10 (high resistance) | 0.17 d ± 0.00 | 1.78 a ± 0.01 | 0.917 c ± 0.01 | 2.08 b ± 0.05 |
6.8 (medium resistance) | 0.09 b ± 0.01 | 3.23 c ± 0.16 | 0.800 b ± 0.07 | 1.71 a ± 0.03 | |
7.1 (sensitive) | 0.00 a ± 0.01 | >24 d | 0.088 a ± 0.01 | >24 c | |
7.15 (medium resistance) | 0.15 c ± 0.07 | 1.98 a,b ± 0.04 | 0.765 b ± 0.01 | 1.68 a ± 0.04 | |
19.27 (high resistance) | 0.14 c ± 0.01 | 2.04 b ± 0.06 | 0.839 c ± 0.03 | 1.96 b ± 0.13 |
Antibiotic | Total E.coli Resistant Isolates | Amplified Gene | E. coli | % |
---|---|---|---|---|
Beta-lactam (ampicillin and amoxicillin) | 156 | blaSHV | 0 | 0 |
blaTEM | 156 | 100 | ||
blaCMY-2 | 0 | 0 | ||
All genes | 0 | 0 | ||
No gene | 0 | 0 | ||
Tetracycline | 176 | tet(A) | 117 | 67 |
tet(B) | 16 | 9 | ||
All genes | 0 | 0 | ||
No gene | 42 | 24 |
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García-Béjar, B.; García de Blas Martín, I.; Arévalo-Villena, M.; Briones Pérez, A. High Prevalence of Antibiotic-Resistant Escherichia coli Isolates from Retail Poultry Products in Spain. Animals 2021, 11, 3197. https://doi.org/10.3390/ani11113197
García-Béjar B, García de Blas Martín I, Arévalo-Villena M, Briones Pérez A. High Prevalence of Antibiotic-Resistant Escherichia coli Isolates from Retail Poultry Products in Spain. Animals. 2021; 11(11):3197. https://doi.org/10.3390/ani11113197
Chicago/Turabian StyleGarcía-Béjar, Beatriz, Izan García de Blas Martín, María Arévalo-Villena, and Ana Briones Pérez. 2021. "High Prevalence of Antibiotic-Resistant Escherichia coli Isolates from Retail Poultry Products in Spain" Animals 11, no. 11: 3197. https://doi.org/10.3390/ani11113197