The potential of bacteriophage for controlling Vibrio parahaemolyticus as in-vitro
Vibrio parahaemolyticus infection is a major threat to Pacific white shrimp aquaculture, causing significant shrimp mortality. Bacteriophages (phages) provide a promising antibiotic alternative due to their specificity towards specific bacteria. This research includes essential steps isolating phag...
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Asosiasi Sains Akuakultur Indonesia
2024-07-01
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| Series: | Jurnal Akuakultur Indonesia |
| Online Access: | https://journal.ipb.ac.id/index.php/jai/article/view/51147 |
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| author | Bagus Ansani Takwin Dinamella Wahjuningrum Widanarni Widanarni Hasan Nasrullah |
| author_facet | Bagus Ansani Takwin Dinamella Wahjuningrum Widanarni Widanarni Hasan Nasrullah |
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Vibrio parahaemolyticus infection is a major threat to Pacific white shrimp aquaculture, causing significant shrimp mortality. Bacteriophages (phages) provide a promising antibiotic alternative due to their specificity towards specific bacteria. This research includes essential steps isolating phages from shrimp pond water and characterizing them, including plaque morphology, spot tests, phage density evaluations, host range assessments, and in vitro studies targeting V. parahaemolyticus. In-vitro tests encompassed six treatments, each with three replicates: negative control (K-), positive control (K+), antibiotic control, phage density of 107 (FB7), phage density of 108 (FB8), and phage density of FB109 (FB9). Phages were successfully isolated from shrimp pond water in Banyuwangi, indicated by plaque formation on double-layer agar media. These phages exhibited morphological characteristics, featuring small, round plaques (d= 0.02-0.5 ± 0.08 cm) with flat, wavy edges and clear plaque. Phage plaques were amplified with an average density of 3.5×109 PFU/mL. Host range tests revealed that only V. parahaemolyticus Situbondo was susceptible to the isolated phages, while five other bacterial strains were not. In bacterial inhibition tests, treatments with different phage densities significantly outperformed the negative control, media control, and antibiotic control. However, the antibiotic control performed better than phage density treatments (0.61 ± 0.01). All phage density treatments effectively reduced V. parahaemolyticus density compared to the negative control, with the most favorable results observed in the FB9 treatment at a density of 109 PFU/mL.
Keywords: Aquaculture, antibiotic, bacteriophage, vannamei shrimp, Vibrio parahaemolyticus
ABSTRAK
Penyakit akibat infeksi Vibrio parahaemolyticus menjadi perhatian utama dalam budidaya udang vaname saat ini karena dapat menyebabkan kematian massal pada udang. Penggunaan bakteriofage (fage) dapat menjadi alternatif solusi untuk menggantikan antibiotik, karena fage bersifat spesifik pada bakteri tertentu. Penelitian ini meliputi beberapa tahapan, yaitu isolasi fage dari beberapa sumber air tambak, karakterisasi fage meliputi morfologi plak fage, spot test, dan uji kisaran inang, serta uji in vitro penghambatan V. parahaemolyticus oleh fage. Uji in vitro dilakukan dengan enam perlakuan dan masing masing tiga ulangan yaitu kontrol negatif (K-), kontrol positif (K+), kontrol antibiotik, kepadatan fage 107 (FB7), kepadatan fage 108 (FB8), dan kepadatan fage 109 (FB9). Penelitian ini berhasil mengisolasi fage yang bersumber dari air tambak udang di Banyuwangi ditandai dengan terbentuknya plak pada media double layer agar, dengan ciri morfologisnya berbentuk titik-titik dan bulat besar (d= 0,02 ± 0,08 cm), tepian rata dan bergelombang serta memiliki warna plak yang bening. Plak fage dapat diperbanyak dengan densitas fage rata-rata 3,5×109 PFU/mL. Untuk uji kisaran inang menggunakan 6 jenis bakteri dengan sumber dan strain yang berbeda, menunjukkan bahwa hanya bakteri V. parahaemolyticus Situbondo yang dapat lisis oleh fage hasil isolasi, sedangkan lima jenis bakteri lainnya tidak. Pada daya hambat bakteri, perlakuan dengan penambahan pada setiap densitas fage berbeda nyata dibandingkan dengan kontrol negatif, kontrol media dan kontrol antibiotik, walaupun kontrol antibiotik lebih baik dari perlakuan densitas fage (0,61 ± 0,01). Semua perlakuan kepadatan fage mampu mengurangi kepadatan V. parahaemolyticus dibandingkan dengan perlakuan kontrol negatif. Hasil terbaik terdapat pada perlakuan FB9 yaitu 109 PFU/mL.
Kata kunci: akuakultur, antibiotik, bakteriofage, udang vaname, Vibrio parahaemolyticus
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| format | Article |
| id | doaj-art-5b41f20aea0649329ff2c9d8c114b86a |
| institution | Kabale University |
| issn | 1412-5269 2354-6700 |
| language | English |
| publishDate | 2024-07-01 |
| publisher | Asosiasi Sains Akuakultur Indonesia |
| record_format | Article |
| series | Jurnal Akuakultur Indonesia |
| spelling | doaj-art-5b41f20aea0649329ff2c9d8c114b86a2025-01-24T04:32:43ZengAsosiasi Sains Akuakultur IndonesiaJurnal Akuakultur Indonesia1412-52692354-67002024-07-0123210.19027/jai.23.2.122-13351147The potential of bacteriophage for controlling Vibrio parahaemolyticus as in-vitroBagus Ansani Takwin0Dinamella Wahjuningrum1Widanarni Widanarni2Hasan Nasrullah3Department of Aquaculture, Faculty of Fisheries and Marine Sciences, IPB University, Bogor, West Java, IndonesiaDepartment of Aquaculture, Faculty of Fisheries and Marine Sciences, IPB University, Bogor, West Java, IndonesiaDepartment of Aquaculture, Faculty of Fisheries and Marine Sciences, IPB University, Bogor, West Java, IndonesiaDepartment of Aquaculture, Faculty of Fisheries and Marine Sciences, IPB University, Bogor, West Java, Indonesia Vibrio parahaemolyticus infection is a major threat to Pacific white shrimp aquaculture, causing significant shrimp mortality. Bacteriophages (phages) provide a promising antibiotic alternative due to their specificity towards specific bacteria. This research includes essential steps isolating phages from shrimp pond water and characterizing them, including plaque morphology, spot tests, phage density evaluations, host range assessments, and in vitro studies targeting V. parahaemolyticus. In-vitro tests encompassed six treatments, each with three replicates: negative control (K-), positive control (K+), antibiotic control, phage density of 107 (FB7), phage density of 108 (FB8), and phage density of FB109 (FB9). Phages were successfully isolated from shrimp pond water in Banyuwangi, indicated by plaque formation on double-layer agar media. These phages exhibited morphological characteristics, featuring small, round plaques (d= 0.02-0.5 ± 0.08 cm) with flat, wavy edges and clear plaque. Phage plaques were amplified with an average density of 3.5×109 PFU/mL. Host range tests revealed that only V. parahaemolyticus Situbondo was susceptible to the isolated phages, while five other bacterial strains were not. In bacterial inhibition tests, treatments with different phage densities significantly outperformed the negative control, media control, and antibiotic control. However, the antibiotic control performed better than phage density treatments (0.61 ± 0.01). All phage density treatments effectively reduced V. parahaemolyticus density compared to the negative control, with the most favorable results observed in the FB9 treatment at a density of 109 PFU/mL. Keywords: Aquaculture, antibiotic, bacteriophage, vannamei shrimp, Vibrio parahaemolyticus ABSTRAK Penyakit akibat infeksi Vibrio parahaemolyticus menjadi perhatian utama dalam budidaya udang vaname saat ini karena dapat menyebabkan kematian massal pada udang. Penggunaan bakteriofage (fage) dapat menjadi alternatif solusi untuk menggantikan antibiotik, karena fage bersifat spesifik pada bakteri tertentu. Penelitian ini meliputi beberapa tahapan, yaitu isolasi fage dari beberapa sumber air tambak, karakterisasi fage meliputi morfologi plak fage, spot test, dan uji kisaran inang, serta uji in vitro penghambatan V. parahaemolyticus oleh fage. Uji in vitro dilakukan dengan enam perlakuan dan masing masing tiga ulangan yaitu kontrol negatif (K-), kontrol positif (K+), kontrol antibiotik, kepadatan fage 107 (FB7), kepadatan fage 108 (FB8), dan kepadatan fage 109 (FB9). Penelitian ini berhasil mengisolasi fage yang bersumber dari air tambak udang di Banyuwangi ditandai dengan terbentuknya plak pada media double layer agar, dengan ciri morfologisnya berbentuk titik-titik dan bulat besar (d= 0,02 ± 0,08 cm), tepian rata dan bergelombang serta memiliki warna plak yang bening. Plak fage dapat diperbanyak dengan densitas fage rata-rata 3,5×109 PFU/mL. Untuk uji kisaran inang menggunakan 6 jenis bakteri dengan sumber dan strain yang berbeda, menunjukkan bahwa hanya bakteri V. parahaemolyticus Situbondo yang dapat lisis oleh fage hasil isolasi, sedangkan lima jenis bakteri lainnya tidak. Pada daya hambat bakteri, perlakuan dengan penambahan pada setiap densitas fage berbeda nyata dibandingkan dengan kontrol negatif, kontrol media dan kontrol antibiotik, walaupun kontrol antibiotik lebih baik dari perlakuan densitas fage (0,61 ± 0,01). Semua perlakuan kepadatan fage mampu mengurangi kepadatan V. parahaemolyticus dibandingkan dengan perlakuan kontrol negatif. Hasil terbaik terdapat pada perlakuan FB9 yaitu 109 PFU/mL. Kata kunci: akuakultur, antibiotik, bakteriofage, udang vaname, Vibrio parahaemolyticus https://journal.ipb.ac.id/index.php/jai/article/view/51147 |
| spellingShingle | Bagus Ansani Takwin Dinamella Wahjuningrum Widanarni Widanarni Hasan Nasrullah The potential of bacteriophage for controlling Vibrio parahaemolyticus as in-vitro Jurnal Akuakultur Indonesia |
| title | The potential of bacteriophage for controlling Vibrio parahaemolyticus as in-vitro |
| title_full | The potential of bacteriophage for controlling Vibrio parahaemolyticus as in-vitro |
| title_fullStr | The potential of bacteriophage for controlling Vibrio parahaemolyticus as in-vitro |
| title_full_unstemmed | The potential of bacteriophage for controlling Vibrio parahaemolyticus as in-vitro |
| title_short | The potential of bacteriophage for controlling Vibrio parahaemolyticus as in-vitro |
| title_sort | potential of bacteriophage for controlling vibrio parahaemolyticus as in vitro |
| url | https://journal.ipb.ac.id/index.php/jai/article/view/51147 |
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