Characterization and molecular detection of pathogenicity and antibiotic resistance genes in Vibrio parahaemolyticus isolated from Pacific white shrimp
ABSTRACT This study aimed to conduct the characterization and molecular detection of the pathogenicity and antibiotic-resistance genes in Vibrio parahaemolyticus, as the causative agent of vibriosis in Pacific white shrimp. The V. parahaemolyticus isolates were collected from the shrimp’s h...
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| Format: | Article |
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Asosiasi Sains Akuakultur Indonesia
2022-07-01
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| Series: | Jurnal Akuakultur Indonesia |
| Online Access: | https://journal.ipb.ac.id/index.php/jai/article/view/42200 |
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| author | Muhammad Arif Mulya Fachriyan Hasmi Pasaribu Usamah Afiff Munti Yuhana |
| author_facet | Muhammad Arif Mulya Fachriyan Hasmi Pasaribu Usamah Afiff Munti Yuhana |
| author_sort | Muhammad Arif Mulya |
| collection | DOAJ |
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ABSTRACT
This study aimed to conduct the characterization and molecular detection of the pathogenicity and antibiotic-resistance genes in Vibrio parahaemolyticus, as the causative agent of vibriosis in Pacific white shrimp. The V. parahaemolyticus isolates were collected from the shrimp’s hepatopancreas, before biochemical test and polymerase chain reaction (PCR) sequencing of the 16S rRNA gene confirmation. The hemolysis test and PCR were applied to detect the presence of virulence genes, namely toxR, thermostable direct haemolysin (tdh), and tdh-related haemolysin (trh). The Kirby-Bauer method was used for characterizing the resistance patterns against ampicillin (AMP), tetracycline (TET), cyprofloxacin (CIP), enrofloxacin (ENR), and chloramphenicol (CHL). The biochemical tests and PCR-16SrRNA gene sequencing confirmed that 12 isolates belonged to V. parahaemolyticus that were further verified by amplification of the toxR gene in 382 bp (100% of the isolates). The alpha hemolysis activity was also confirmed by the amplicon of 199 bp in all isolates. All V. parahaemolyticus isolates showed their resistance to AMP and 42% of the isolates were TET-resistant. However, no resistance was shown to CIP, ENR, and CHL. The PCR-based analysis resulted a detectable resistance gene of ampC (42% of the isolates) and tetB (83% of the isolates).
Keywords: antibiotics, shrimp, resistance, virulency, Vibrio parahaemolyticus
ABSTRAK
Penelitian ini bertujuan untuk melakukan karakterisasi dan deteksi molekular dari gen patogenisitas dan resistansi antibiotik pada Vibrio parahaemolyticus, agen penyebab vibriosis pada udang vaname. Isolat V. parahaemolyticus dikoleksi dari hepatopankreas, diuji secara biokimiawi dan selanjutnya dikonfirmasi dengan polymerase chain reaction (PCR)-sekuensing dari gen 16S rRNA. Tes hemolisis dan metode PCR diterapkan untuk mendeteksi keberadaan gen virulensi toxR, thermostable direct haemolysin (tdh) and tdh-related haemolysin (trh). Metode Kirby Bauer digunakan untuk karakterisasi pola resistansi terhadap ampisilin (AMP), tetrasiklin (TET), kloramfenikol (CHL), siprofloksasin (CIP) dan enrofloksasin (ENR). Uji biokimia dan sekuensing gen PCR-16SrRNA memastikan bahwa 12 isolat adalah V. parahaemolyticus yang selanjutnya diverifikasi dengan amplifikasi gen toxR berukuran 382 bp (100% isolat). Aktivitas alfa hemolisis juga dikonfirmasi dengan amplikon PCR (199 bp) di semua isolat. Seluruh isolat V. parahaemolyticus menunjukkan resistansinya terhadap AMP, 42% resistan TET, tidak ada resistansi yang ditunjukkan pada CIP, ENR dan CHL. Analisis berbasis PCR menghasilkan gen resistan yang terdeteksi dari gen ampC (42% isolat) dan gen tetB (83% isolat).
Kata kunci: Antibiotik, udang, resistansi, virulensi, Vibrio parahaemolyticus
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| format | Article |
| id | doaj-art-f2c6f0d0c8de4b2ab71b20caecb1866e |
| institution | Kabale University |
| issn | 1412-5269 2354-6700 |
| language | English |
| publishDate | 2022-07-01 |
| publisher | Asosiasi Sains Akuakultur Indonesia |
| record_format | Article |
| series | Jurnal Akuakultur Indonesia |
| spelling | doaj-art-f2c6f0d0c8de4b2ab71b20caecb1866e2025-01-24T17:56:16ZengAsosiasi Sains Akuakultur IndonesiaJurnal Akuakultur Indonesia1412-52692354-67002022-07-0121110.19027/jai.21.1.81-9242200Characterization and molecular detection of pathogenicity and antibiotic resistance genes in Vibrio parahaemolyticus isolated from Pacific white shrimpMuhammad Arif Mulya0Fachriyan Hasmi Pasaribu1Usamah Afiff2Munti Yuhana3Aquaculture Production Engineering and Management Study Program, Vocational School, IPB UniversityMedical Microbiology Study Program, Postgraduate School, IPB UniversityDepartment of Animal Disease and Veterinary Public Health, IPB UniversityDepartment of Aquaculture, Faculty of Fisheries and Marine Sciences, IPB University, Bogor West Java 16680 ABSTRACT This study aimed to conduct the characterization and molecular detection of the pathogenicity and antibiotic-resistance genes in Vibrio parahaemolyticus, as the causative agent of vibriosis in Pacific white shrimp. The V. parahaemolyticus isolates were collected from the shrimp’s hepatopancreas, before biochemical test and polymerase chain reaction (PCR) sequencing of the 16S rRNA gene confirmation. The hemolysis test and PCR were applied to detect the presence of virulence genes, namely toxR, thermostable direct haemolysin (tdh), and tdh-related haemolysin (trh). The Kirby-Bauer method was used for characterizing the resistance patterns against ampicillin (AMP), tetracycline (TET), cyprofloxacin (CIP), enrofloxacin (ENR), and chloramphenicol (CHL). The biochemical tests and PCR-16SrRNA gene sequencing confirmed that 12 isolates belonged to V. parahaemolyticus that were further verified by amplification of the toxR gene in 382 bp (100% of the isolates). The alpha hemolysis activity was also confirmed by the amplicon of 199 bp in all isolates. All V. parahaemolyticus isolates showed their resistance to AMP and 42% of the isolates were TET-resistant. However, no resistance was shown to CIP, ENR, and CHL. The PCR-based analysis resulted a detectable resistance gene of ampC (42% of the isolates) and tetB (83% of the isolates). Keywords: antibiotics, shrimp, resistance, virulency, Vibrio parahaemolyticus ABSTRAK Penelitian ini bertujuan untuk melakukan karakterisasi dan deteksi molekular dari gen patogenisitas dan resistansi antibiotik pada Vibrio parahaemolyticus, agen penyebab vibriosis pada udang vaname. Isolat V. parahaemolyticus dikoleksi dari hepatopankreas, diuji secara biokimiawi dan selanjutnya dikonfirmasi dengan polymerase chain reaction (PCR)-sekuensing dari gen 16S rRNA. Tes hemolisis dan metode PCR diterapkan untuk mendeteksi keberadaan gen virulensi toxR, thermostable direct haemolysin (tdh) and tdh-related haemolysin (trh). Metode Kirby Bauer digunakan untuk karakterisasi pola resistansi terhadap ampisilin (AMP), tetrasiklin (TET), kloramfenikol (CHL), siprofloksasin (CIP) dan enrofloksasin (ENR). Uji biokimia dan sekuensing gen PCR-16SrRNA memastikan bahwa 12 isolat adalah V. parahaemolyticus yang selanjutnya diverifikasi dengan amplifikasi gen toxR berukuran 382 bp (100% isolat). Aktivitas alfa hemolisis juga dikonfirmasi dengan amplikon PCR (199 bp) di semua isolat. Seluruh isolat V. parahaemolyticus menunjukkan resistansinya terhadap AMP, 42% resistan TET, tidak ada resistansi yang ditunjukkan pada CIP, ENR dan CHL. Analisis berbasis PCR menghasilkan gen resistan yang terdeteksi dari gen ampC (42% isolat) dan gen tetB (83% isolat). Kata kunci: Antibiotik, udang, resistansi, virulensi, Vibrio parahaemolyticus https://journal.ipb.ac.id/index.php/jai/article/view/42200 |
| spellingShingle | Muhammad Arif Mulya Fachriyan Hasmi Pasaribu Usamah Afiff Munti Yuhana Characterization and molecular detection of pathogenicity and antibiotic resistance genes in Vibrio parahaemolyticus isolated from Pacific white shrimp Jurnal Akuakultur Indonesia |
| title | Characterization and molecular detection of pathogenicity and antibiotic resistance genes in Vibrio parahaemolyticus isolated from Pacific white shrimp |
| title_full | Characterization and molecular detection of pathogenicity and antibiotic resistance genes in Vibrio parahaemolyticus isolated from Pacific white shrimp |
| title_fullStr | Characterization and molecular detection of pathogenicity and antibiotic resistance genes in Vibrio parahaemolyticus isolated from Pacific white shrimp |
| title_full_unstemmed | Characterization and molecular detection of pathogenicity and antibiotic resistance genes in Vibrio parahaemolyticus isolated from Pacific white shrimp |
| title_short | Characterization and molecular detection of pathogenicity and antibiotic resistance genes in Vibrio parahaemolyticus isolated from Pacific white shrimp |
| title_sort | characterization and molecular detection of pathogenicity and antibiotic resistance genes in vibrio parahaemolyticus isolated from pacific white shrimp |
| url | https://journal.ipb.ac.id/index.php/jai/article/view/42200 |
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