Evaluation of single and multispecies probiotic applications for the prevention of Motile Aeromonads Septicaemia in gourami, Osphronemus gourami

Abstract

The aim of this study was to evaluate single and multispecies probiotic applications for the prevention of Motile Aeromonads Septicaemia (MAS) disease in gourami, Osphronemus gourami. The experiment consisted of the in vitro inhibition test and the in vivo application of probiotics in gourami. The in vivo assay, consisted of five treatments and five replicates, namely: negative control (K-); positive control (K+); (B) fish fed with  supplementation of 1% (v/w) probiotic Bacillus NP5 Rif^R cells and challenged with Aeromonas hydrophila Rif^R; (L) fish fed with supplemention of probiotic Lactobacillus plantarum Cip^R 1% (v/w); challenged with A. hydrophila Rif^R; (BL) fish fed with mixture supplementation of Bacillus NP5 Rif^R 0.5% (v/w)+L. plantarum Cip^R 0.5%; infected with A. hydrophila Rif^R. Fish (29.57 ± 1.00 g) were reared in a 45 L volume aquaria with a rearing density of 10 fish/m³ for 50 days, with   feeding trial three times a day. The challenge test was conducted by intramuscular injection with pathogenic A. hydrophila Rif^R cells (10⁶ CFU/mL) on day 41^st. In vitro test results showed that single and multispecies probiotics significantly (P<0.05) inhibited A.hydrophila Rif^R. In vivo experiment showed that probiotic supplementation treatments improved the growth performance, and microbiota diversity in the gut.  The immune responses, fish resistance to A. hydrophila Rif^R and gourami survival rate in all treatments of supplemented feed were significantly higher compared to the positive control. The best treatment, multispecies probiotics significantly (P<0.05) improved the survival of gourami 96.67% post infection with A. hydrophila Rif^R.

Keywords: A. hydrophila, Bacillus NP5, L. plantarum, O. gourami, probiotic

ABSTRAK

Penelitian ini bertujuan untuk mengevaluasi aplikasi probiotik tunggal dan multispesies dalam pencegahan penyakit Motile Aeromonad Septicaemia (MAS) yang disebabkan oleh Aeromonas hydrophila pada ikan gurami (Osphronemus gourami). Penelitian terdiri atas uji penghambatan in vitro bakteri probiotik tunggal dan multispesies terhadap A. hydrophila Rif^R, dan uji in vivo aplikasi probiotik untuk gurami. Uji in vivo, terdiri dari lima perlakuan dan lima ulangan, yaitu: kontrol negatif (K-) kontrol positif (K+); (B) ikan dengan pakan suplementasi Bacillus NP5 Rif^R 1% dan diinfeksi A. hydrophila Rif^R; (L) pakan komersial dengan penambahan probiotik L. plantarum Cip^R 1% dan infeksi A. hydrophila Rif^R; (BL) ikan dengan suplementasi campuran Bacillus NP5 Rif^R 0.5% dan L. plantarum Cip^R 0.5% serta diinfeksi A. hydrophila Rif^R. Benih gurami (29.57 ± 1.00 g) dipelihara di dalam akuarium bervolume 45 L dengan kepadatan 10 ekor/m³ selama 50 hari, dengan pakan bersuplemen probiotik 3 kali sehari. Uji tantang dilakukan dengan menginjeksikan suspensi sel patogen A. hydrophila Rif^R (10⁶ CFU/mL) secara intramuscular pada hari ke 41. Hasil uji in vitro menunjukkan bahwa probiotik tunggal dan multispesies dapat menghambat pertumbuhan sel A.hydrophila Rif^R di organ hati dan ginjal secara signifikan (P<0.05). Hasil uji in vivo menunjukkan perlakuan aplikasi probiotik tunggal dan multispesies mampu meningkatkan kinerja pertumbuhan, keragaman mikrobiota usus. Respons imunitas, resistansi terhadap A. hydrophila Rif^R dan kelangsungan hidup gurami di semua perlakuan pakan bersuplemen probiotik secara signifikan lebih tinggi dibandingkan kontrol positif.  Perlakuan terbaik adalah probiotik multispesies meningkatkan kelangsungan hidup ikan sebesar 96.67% secara signifikan (P<0.05) pascainfeksi A. hydrophila Rif^R.

Kata kunci: A. hydrophila, Bacillus sp. NP5, L. plantarum, O. goramy, probiotik