SKRINING DAN IDENTIFIKASI BAKTERI ANTI QUORUM SENSING ASAL TAMBAK UDANG VANAME PENGHAMBAT VIRULENSI Vibrio parahaemolyticus

Abstract

Vibriosis dapat dicegah dan dikendalikan dengan memanfaatkan mekanisme anti quorum sensing (AQS). Salah satu strategi anti quorum sensing dalam menghambat ekspresi faktor virulen dari Vibrio parahaemolyticus yaitu dengan mendegradasi sinyal komunikasi sel bakteri menggunakan AHL laktonase. Penelitian ini bertujuan untuk menyeleksi dan mengindentifikasi bakteri penghasil AHL laktonase yang berpotensi mampu menghambat virulensi bakteri patogen V. parahaemolyticus. Isolasi bakteri dilakukan dari sampel saluran pencernaan udang vaname, air, dan sedimen tambak. Sebanyak 18 dari 111 isolat yang diisolasi menunjukkan adanya aktivitas AQS terhadap bioindikator Chromobacterium violaceum. Hasil uji patogenitas secara in vitro pada agar darah didapatkan tiga isolat yang tidak menunjukkan aktivitas hemolisis yaitu B5, K4, dan S12. Hasil konfirmasi dan analisis gen aiiA menggunakan Blast-X menunjukkan bahwa isolat B5 dan S12 memiliki kesamaan dengan AHL laktonase pada Bacillus cereus, sedangkan K4 memiliki similaritas dengan AHL laktonase pada multispesies Bacillus sp. Hasil pensejajaran sekuen gen 16S rRNA ketiga isolat tersebut dengan data pada GenBank, teridentifikasi sebagai Bacillus siamensis (B5), Bacillus cereus (K4), dan Bacillus amyloliquefaciens (S12). Berdasarkan hasil uji antagonis dan uji kultur bersama disimpulkan bahwa isolat K4 bekerja dengan mekanisme AQS sedangkan isolat B5 dan S12 diduga berjalan dua mekanisme secara bersama yaitu antibiosis dan anti quorum sensing. Hasil penelitian ini menunjukkan bahwa ketiga isolat tersebut memiliki potensi sebagai kandidat agen biokontrol pada akuakultur sehingga perlu dilakukan uji lanjutan.

Vibriosis can be prevented and controlled by utilizing the anti-quorum sensing (AQS) mechanism. One of the anti-quorum sensing mechanisms to inhibit the expression of virulent factors of Vibrio parahaemolyticus is by degrading the quorum sensing communication signals using AHL lactonase. The study aimed to select and identify AHL lactonase-producing bacteria that have the potentials to inhibit the virulence of V. parahaemolyticus. Several batches of bacteria were isolated from the digestive tract of vannamei shrimp, water, and sediment of shrimp ponds. There were 18 out of 111 isolates that showed AQS activity against Chromobacterium violaceum used as a bioindicator. In vitro pathogenicity test on blood agar showed that B5, K4, and S12 isolates showed gamma hemolysis activity. The results of confirmation and analysis of aiiA genes using Blast-X showed that B5 and S12 isolates have AHL lactonase similarities with Bacillus cereus, whereas K4 has similarities with multispecies Bacillus sp. Alignment results of the 16S rRNA gene sequences with GenBank data showed that B5, K4, and S12 isolates were identified as Bacillus siamensis, Bacillus cereus, and Bacillus amyloliquefaciens, respectively. The follow up antagonistic and coculture tests revealed that K4 uses the AQS mechanism, while B5 and S12 likely use antibiotic mechanism and anti quorum sensing to inhibit the virulent expression of V. parahaemolyticus. This study concludes that the three isolates have the potential to be used as biocontrol agents in brackishwater aquaculture. Further research is needed to determine the pathogenicity of AQS bacteria to vannamei shrimp and the effective concentration of AQS bacteria to inhibit the virulence of V. parahaemolyticus to vannamei shrimp by in vivo treatment.