Detection and Phylogenetic Analysis of Extended-Spectrum β-Lactamase (ESBL)-Genetic Determinants in Gram-Negative Fecal-Microbiota of Wild Birds and Chicken Originated at Trimmu Barrage
Extended-spectrum β-lactamases (ESBL) give rise to resistance against penicillin and cephalosporin antibiotics in multiple bacterial species. The present study was conducted to map genetic determinants and related attributes of ESBL-producing bacteria in three wild aquatic bird species and chickens...
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| Format: | Article |
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MDPI AG
2023-08-01
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| Series: | Antibiotics |
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| Online Access: | https://www.mdpi.com/2079-6382/12/9/1376 |
| _version_ | 1797581564637347840 |
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| author | Muhammad Adnan Saeed Aman Ullah Khan Syed Ehtisham-ul-Haque Usman Waheed Muhammad Fiaz Qamar Aziz ur Rehman Amar Nasir Muhammad Arfan Zaman Muhammad Kashif Jean-Paul Gonzalez Hosny El-Adawy |
| author_facet | Muhammad Adnan Saeed Aman Ullah Khan Syed Ehtisham-ul-Haque Usman Waheed Muhammad Fiaz Qamar Aziz ur Rehman Amar Nasir Muhammad Arfan Zaman Muhammad Kashif Jean-Paul Gonzalez Hosny El-Adawy |
| author_sort | Muhammad Adnan Saeed |
| collection | DOAJ |
| description | Extended-spectrum β-lactamases (ESBL) give rise to resistance against penicillin and cephalosporin antibiotics in multiple bacterial species. The present study was conducted to map genetic determinants and related attributes of ESBL-producing bacteria in three wild aquatic bird species and chickens at the “Trimmu Barrage” in district Jhang, Punjab province, Pakistan. To study the prevalence of ESBL-producing bacteria, a total of 280 representative samples were collected from wild bird species; cattle egrets (<i>Bubulcus ibis</i>), little egrets (<i>Egretta garzetta</i>) and common teals (<i>Anas crecca</i>) as well as from indigenous chickens (<i>Gallus gallus domesticus</i>) originating from a local wet market. The isolates were confirmed as ESBL producers using a double disc synergy test (DDST) and bacterial species were identified using API-20E and 20NE strips. A polymerase chain reaction (PCR) was used to detect ESBL genetic determinants and for genus identification via 16S rRNA gene amplification. A phenotypic antimicrobial susceptibility test was performed for ESBL-producing isolates against 12 clinically relevant antibiotics using the Kirby–Bauer disk diffusion susceptibility test. A phylogenetic tree was constructed for the sequence data obtained in this study and comparative sequence data obtained from GenBank. The overall prevalence of ESBL-producing bacteria was 34.64% (97/280). The highest percentage (44.28%; 31/70) of ESBL-producing bacteria was recovered from chickens (<i>Gallus gallus domesticus</i>), followed by little egrets (<i>Egretta garzetta</i>) (41.43%; 29/70), common teal (<i>Anas crecca</i>) (28.57%; 20/70) and cattle egrets (<i>Bubulcus ibis</i>) (24.28%; 17/70). Five different ESBL-producing bacteria were identified biochemically and confirmed via 16S rRNA gene sequencing, which included <i>Escherichia coli</i> (72; 74.23%), <i>Enterobacter cloacae</i> (11; 11.34%), <i>Klebsiella pneumoniae</i> (8; 8.25%), <i>Salmonella enterica</i> (4; 4.12%) and <i>Pseudomonas aeruginosa</i> (2; 2.06%). Based on PCR, the frequency of obtained ESBL genes in 97 isolates was <i>bla</i><sub>CTX-M</sub> (51.55%), <i>bla</i><sub>TEM</sub> (20.62%), <i>bla</i><sub>OXA</sub> (6.18%) and <i>bla</i><sub>SHV</sub> (2.06%). In addition, gene combinations <i>bla</i><sub>CTX-M</sub> + <i>bla</i><sub>TEM</sub>, <i>bla</i><sub>TEM</sub> + <i>bla</i><sub>OXA</sub> and <i>bla</i><sub>CTX-M</sub> + <i>bla</i><sub>SHV</sub> were also detected in 16.49%, 2.06% and 1.03% of isolates, respectively. The ESBL gene variation was significant (<i>p</i> = 0.02) in different bacterial species while non-significant in relation to different bird species (<i>p</i> = 0.85). Phylogenetic analysis of amino acid sequence data confirmed the existence of CTX-M-15 and TEM betalactamases. The average susceptibility of the antibiotics panel used was lowest for both <i>Klebsiella pneumoniae</i> (62.5% ± 24.42) and <i>Salmonella enterica</i> (62.5% ± 31.08) as compared to <i>Enterobacter cloacae</i> (65.90% ± 21.62), <i>Pseudomonas aeruginosa</i> (70.83% ± 33.42) and <i>Escherichia coli</i> (73.83% ± 26.19). This study provides insight into the role of aquatic wild birds as reservoirs of ESBL-producing bacteria at Trimmu Barrage, Punjab, Pakistan. Hence, active bio-surveillance and environment preservation actions are necessitated to curb antimicrobial resistance. |
| first_indexed | 2024-03-10T23:06:39Z |
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| id | doaj.art-95c1240370124574a83de7781fe40195 |
| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-03-10T23:06:39Z |
| publishDate | 2023-08-01 |
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| series | Antibiotics |
| spelling | doaj.art-95c1240370124574a83de7781fe401952023-11-19T09:16:55ZengMDPI AGAntibiotics2079-63822023-08-01129137610.3390/antibiotics12091376Detection and Phylogenetic Analysis of Extended-Spectrum β-Lactamase (ESBL)-Genetic Determinants in Gram-Negative Fecal-Microbiota of Wild Birds and Chicken Originated at Trimmu BarrageMuhammad Adnan Saeed0Aman Ullah Khan1Syed Ehtisham-ul-Haque2Usman Waheed3Muhammad Fiaz Qamar4Aziz ur Rehman5Amar Nasir6Muhammad Arfan Zaman7Muhammad Kashif8Jean-Paul Gonzalez9Hosny El-Adawy10Department of Pathobiology, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang 35200, PakistanDepartment of Pathobiology, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang 35200, PakistanDepartment of Pathobiology, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang 35200, PakistanDepartment of Pathobiology, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang 35200, PakistanDepartment of Pathobiology, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang 35200, PakistanDepartment of Pathobiology, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang 35200, PakistanDepartment of Clinical Sciences, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang 35200, PakistanDepartment of Pathobiology, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang 35200, PakistanDepartment of Clinical Sciences, University of Veterinary and Animal Sciences, Lahore, CVAS Campus, 12-Km Chiniot Road, Jhang 35200, PakistanDepartment of Microbiology & Immunology, School of Medicine, Georgetown University, Washington, DC 20057, USAInstitute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, 07743 Jena, GermanyExtended-spectrum β-lactamases (ESBL) give rise to resistance against penicillin and cephalosporin antibiotics in multiple bacterial species. The present study was conducted to map genetic determinants and related attributes of ESBL-producing bacteria in three wild aquatic bird species and chickens at the “Trimmu Barrage” in district Jhang, Punjab province, Pakistan. To study the prevalence of ESBL-producing bacteria, a total of 280 representative samples were collected from wild bird species; cattle egrets (<i>Bubulcus ibis</i>), little egrets (<i>Egretta garzetta</i>) and common teals (<i>Anas crecca</i>) as well as from indigenous chickens (<i>Gallus gallus domesticus</i>) originating from a local wet market. The isolates were confirmed as ESBL producers using a double disc synergy test (DDST) and bacterial species were identified using API-20E and 20NE strips. A polymerase chain reaction (PCR) was used to detect ESBL genetic determinants and for genus identification via 16S rRNA gene amplification. A phenotypic antimicrobial susceptibility test was performed for ESBL-producing isolates against 12 clinically relevant antibiotics using the Kirby–Bauer disk diffusion susceptibility test. A phylogenetic tree was constructed for the sequence data obtained in this study and comparative sequence data obtained from GenBank. The overall prevalence of ESBL-producing bacteria was 34.64% (97/280). The highest percentage (44.28%; 31/70) of ESBL-producing bacteria was recovered from chickens (<i>Gallus gallus domesticus</i>), followed by little egrets (<i>Egretta garzetta</i>) (41.43%; 29/70), common teal (<i>Anas crecca</i>) (28.57%; 20/70) and cattle egrets (<i>Bubulcus ibis</i>) (24.28%; 17/70). Five different ESBL-producing bacteria were identified biochemically and confirmed via 16S rRNA gene sequencing, which included <i>Escherichia coli</i> (72; 74.23%), <i>Enterobacter cloacae</i> (11; 11.34%), <i>Klebsiella pneumoniae</i> (8; 8.25%), <i>Salmonella enterica</i> (4; 4.12%) and <i>Pseudomonas aeruginosa</i> (2; 2.06%). Based on PCR, the frequency of obtained ESBL genes in 97 isolates was <i>bla</i><sub>CTX-M</sub> (51.55%), <i>bla</i><sub>TEM</sub> (20.62%), <i>bla</i><sub>OXA</sub> (6.18%) and <i>bla</i><sub>SHV</sub> (2.06%). In addition, gene combinations <i>bla</i><sub>CTX-M</sub> + <i>bla</i><sub>TEM</sub>, <i>bla</i><sub>TEM</sub> + <i>bla</i><sub>OXA</sub> and <i>bla</i><sub>CTX-M</sub> + <i>bla</i><sub>SHV</sub> were also detected in 16.49%, 2.06% and 1.03% of isolates, respectively. The ESBL gene variation was significant (<i>p</i> = 0.02) in different bacterial species while non-significant in relation to different bird species (<i>p</i> = 0.85). Phylogenetic analysis of amino acid sequence data confirmed the existence of CTX-M-15 and TEM betalactamases. The average susceptibility of the antibiotics panel used was lowest for both <i>Klebsiella pneumoniae</i> (62.5% ± 24.42) and <i>Salmonella enterica</i> (62.5% ± 31.08) as compared to <i>Enterobacter cloacae</i> (65.90% ± 21.62), <i>Pseudomonas aeruginosa</i> (70.83% ± 33.42) and <i>Escherichia coli</i> (73.83% ± 26.19). This study provides insight into the role of aquatic wild birds as reservoirs of ESBL-producing bacteria at Trimmu Barrage, Punjab, Pakistan. Hence, active bio-surveillance and environment preservation actions are necessitated to curb antimicrobial resistance.https://www.mdpi.com/2079-6382/12/9/1376ESBLmicrobiotawild birdsaquatic birdsTrimmu Barragephylogenetics |
| spellingShingle | Muhammad Adnan Saeed Aman Ullah Khan Syed Ehtisham-ul-Haque Usman Waheed Muhammad Fiaz Qamar Aziz ur Rehman Amar Nasir Muhammad Arfan Zaman Muhammad Kashif Jean-Paul Gonzalez Hosny El-Adawy Detection and Phylogenetic Analysis of Extended-Spectrum β-Lactamase (ESBL)-Genetic Determinants in Gram-Negative Fecal-Microbiota of Wild Birds and Chicken Originated at Trimmu Barrage Antibiotics ESBL microbiota wild birds aquatic birds Trimmu Barrage phylogenetics |
| title | Detection and Phylogenetic Analysis of Extended-Spectrum β-Lactamase (ESBL)-Genetic Determinants in Gram-Negative Fecal-Microbiota of Wild Birds and Chicken Originated at Trimmu Barrage |
| title_full | Detection and Phylogenetic Analysis of Extended-Spectrum β-Lactamase (ESBL)-Genetic Determinants in Gram-Negative Fecal-Microbiota of Wild Birds and Chicken Originated at Trimmu Barrage |
| title_fullStr | Detection and Phylogenetic Analysis of Extended-Spectrum β-Lactamase (ESBL)-Genetic Determinants in Gram-Negative Fecal-Microbiota of Wild Birds and Chicken Originated at Trimmu Barrage |
| title_full_unstemmed | Detection and Phylogenetic Analysis of Extended-Spectrum β-Lactamase (ESBL)-Genetic Determinants in Gram-Negative Fecal-Microbiota of Wild Birds and Chicken Originated at Trimmu Barrage |
| title_short | Detection and Phylogenetic Analysis of Extended-Spectrum β-Lactamase (ESBL)-Genetic Determinants in Gram-Negative Fecal-Microbiota of Wild Birds and Chicken Originated at Trimmu Barrage |
| title_sort | detection and phylogenetic analysis of extended spectrum β lactamase esbl genetic determinants in gram negative fecal microbiota of wild birds and chicken originated at trimmu barrage |
| topic | ESBL microbiota wild birds aquatic birds Trimmu Barrage phylogenetics |
| url | https://www.mdpi.com/2079-6382/12/9/1376 |
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