Antimicrobial Susceptibility of Commensal <i>Escherichia coli</i> from Pig Fecal Samples and Enhanced Sensitivity for Direct Detection of the <i>bla</i><sub>CTX-M</sub> Gene by Nested PCR
The commensal <i>Escherichia coli</i> in the gut of pigs is a major reservoir of antimicrobial resistance and can result in possible transmission to humans through the food chain. Direct detection of <i>E. coli</i> from fecal samples is challenging and can be used as a bioind...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-09-01
|
| Series: | Animals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2615/14/18/2630 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850258720202686464 |
|---|---|
| author | Nutchaba Suchanta Naeem Ullah Pitak Santanirand Nutthee Am-In Nuntaree Chaichanawongsaroj |
| author_facet | Nutchaba Suchanta Naeem Ullah Pitak Santanirand Nutthee Am-In Nuntaree Chaichanawongsaroj |
| author_sort | Nutchaba Suchanta |
| collection | DOAJ |
| description | The commensal <i>Escherichia coli</i> in the gut of pigs is a major reservoir of antimicrobial resistance and can result in possible transmission to humans through the food chain. Direct detection of <i>E. coli</i> from fecal samples is challenging and can be used as a bioindicator of antimicrobial resistance. This study aimed to compare the antimicrobial susceptibility profiles in commensal <i>E. coli</i> from antibiotic- and nonantibiotic-using pig farms and developed the direct detection of ESBL genes in pig fecal samples using nested PCR (nPCR) and multiplex PCR (mPCR) techniques. All direct genotypic results were validated with the results of PCR sequencing of isolated <i>E. coli</i> colonies. The ESBL-producing <i>E. coli</i> were found in 98.6% (145 isolates) and 96.6% (144 isolates) of antibiotic-using and nonantibiotic-using farms, respectively, predominantly CTX-M-55. The nPCR decreased the limit of detection (LOD) from sPCR about 100 times, and the lower LODs of 10<sup>2</sup>, 10<sup>1</sup>, and 1 CFU/mL were reached after incubating samples in an enrichment medium for 2, 4, and 8 h, respectively. The mPCR, sPCR, and nPCR techniques showed sensitivities of 30.15%, 69.85%, and 91.91%, respectively, compared to PCR sequencing. The stability and recycling of ESBL genes were independent of antibiotic usage in commensal <i>E. coli</i> originating in pig farms. |
| format | Article |
| id | doaj-art-82345c45851f4b6eb3fac91ffbc9991c |
| institution | OA Journals |
| issn | 2076-2615 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj-art-82345c45851f4b6eb3fac91ffbc9991c2025-08-20T01:56:04ZengMDPI AGAnimals2076-26152024-09-011418263010.3390/ani14182630Antimicrobial Susceptibility of Commensal <i>Escherichia coli</i> from Pig Fecal Samples and Enhanced Sensitivity for Direct Detection of the <i>bla</i><sub>CTX-M</sub> Gene by Nested PCRNutchaba Suchanta0Naeem Ullah1Pitak Santanirand2Nutthee Am-In3Nuntaree Chaichanawongsaroj4Center of Excellence for Innovative Diagnosis of Antimicrobial Resistance, Department of Transfusion Medicine and Clinical Microbiology, Faculty of Allied Health Sciences, Chulalongkorn University, Pathumwan, Bangkok 10330, ThailandCenter of Excellence for Innovative Diagnosis of Antimicrobial Resistance, Department of Transfusion Medicine and Clinical Microbiology, Faculty of Allied Health Sciences, Chulalongkorn University, Pathumwan, Bangkok 10330, ThailandDepartment of Pathology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok 10400, ThailandDepartment of Obstetrics Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Pathumwan, Bangkok 10330, ThailandCenter of Excellence for Innovative Diagnosis of Antimicrobial Resistance, Department of Transfusion Medicine and Clinical Microbiology, Faculty of Allied Health Sciences, Chulalongkorn University, Pathumwan, Bangkok 10330, ThailandThe commensal <i>Escherichia coli</i> in the gut of pigs is a major reservoir of antimicrobial resistance and can result in possible transmission to humans through the food chain. Direct detection of <i>E. coli</i> from fecal samples is challenging and can be used as a bioindicator of antimicrobial resistance. This study aimed to compare the antimicrobial susceptibility profiles in commensal <i>E. coli</i> from antibiotic- and nonantibiotic-using pig farms and developed the direct detection of ESBL genes in pig fecal samples using nested PCR (nPCR) and multiplex PCR (mPCR) techniques. All direct genotypic results were validated with the results of PCR sequencing of isolated <i>E. coli</i> colonies. The ESBL-producing <i>E. coli</i> were found in 98.6% (145 isolates) and 96.6% (144 isolates) of antibiotic-using and nonantibiotic-using farms, respectively, predominantly CTX-M-55. The nPCR decreased the limit of detection (LOD) from sPCR about 100 times, and the lower LODs of 10<sup>2</sup>, 10<sup>1</sup>, and 1 CFU/mL were reached after incubating samples in an enrichment medium for 2, 4, and 8 h, respectively. The mPCR, sPCR, and nPCR techniques showed sensitivities of 30.15%, 69.85%, and 91.91%, respectively, compared to PCR sequencing. The stability and recycling of ESBL genes were independent of antibiotic usage in commensal <i>E. coli</i> originating in pig farms.https://www.mdpi.com/2076-2615/14/18/2630<i>E. coli</i>ESBLsantimicrobial resistance<i>bla</i><sub>CTX-M</sub> genenPCRpig feces |
| spellingShingle | Nutchaba Suchanta Naeem Ullah Pitak Santanirand Nutthee Am-In Nuntaree Chaichanawongsaroj Antimicrobial Susceptibility of Commensal <i>Escherichia coli</i> from Pig Fecal Samples and Enhanced Sensitivity for Direct Detection of the <i>bla</i><sub>CTX-M</sub> Gene by Nested PCR Animals <i>E. coli</i> ESBLs antimicrobial resistance <i>bla</i><sub>CTX-M</sub> gene nPCR pig feces |
| title | Antimicrobial Susceptibility of Commensal <i>Escherichia coli</i> from Pig Fecal Samples and Enhanced Sensitivity for Direct Detection of the <i>bla</i><sub>CTX-M</sub> Gene by Nested PCR |
| title_full | Antimicrobial Susceptibility of Commensal <i>Escherichia coli</i> from Pig Fecal Samples and Enhanced Sensitivity for Direct Detection of the <i>bla</i><sub>CTX-M</sub> Gene by Nested PCR |
| title_fullStr | Antimicrobial Susceptibility of Commensal <i>Escherichia coli</i> from Pig Fecal Samples and Enhanced Sensitivity for Direct Detection of the <i>bla</i><sub>CTX-M</sub> Gene by Nested PCR |
| title_full_unstemmed | Antimicrobial Susceptibility of Commensal <i>Escherichia coli</i> from Pig Fecal Samples and Enhanced Sensitivity for Direct Detection of the <i>bla</i><sub>CTX-M</sub> Gene by Nested PCR |
| title_short | Antimicrobial Susceptibility of Commensal <i>Escherichia coli</i> from Pig Fecal Samples and Enhanced Sensitivity for Direct Detection of the <i>bla</i><sub>CTX-M</sub> Gene by Nested PCR |
| title_sort | antimicrobial susceptibility of commensal i escherichia coli i from pig fecal samples and enhanced sensitivity for direct detection of the i bla i sub ctx m sub gene by nested pcr |
| topic | <i>E. coli</i> ESBLs antimicrobial resistance <i>bla</i><sub>CTX-M</sub> gene nPCR pig feces |
| url | https://www.mdpi.com/2076-2615/14/18/2630 |
| work_keys_str_mv | AT nutchabasuchanta antimicrobialsusceptibilityofcommensaliescherichiacoliifrompigfecalsamplesandenhancedsensitivityfordirectdetectionoftheiblaisubctxmsubgenebynestedpcr AT naeemullah antimicrobialsusceptibilityofcommensaliescherichiacoliifrompigfecalsamplesandenhancedsensitivityfordirectdetectionoftheiblaisubctxmsubgenebynestedpcr AT pitaksantanirand antimicrobialsusceptibilityofcommensaliescherichiacoliifrompigfecalsamplesandenhancedsensitivityfordirectdetectionoftheiblaisubctxmsubgenebynestedpcr AT nuttheeamin antimicrobialsusceptibilityofcommensaliescherichiacoliifrompigfecalsamplesandenhancedsensitivityfordirectdetectionoftheiblaisubctxmsubgenebynestedpcr AT nuntareechaichanawongsaroj antimicrobialsusceptibilityofcommensaliescherichiacoliifrompigfecalsamplesandenhancedsensitivityfordirectdetectionoftheiblaisubctxmsubgenebynestedpcr |