Functional Characterization of Probiotic Potential of Novel Pigmented Bacterial Strains for Aquaculture Applications
The bioprospecting proficient of novel pigmented probiotic strains with respect to aquaculture industry was unexplored hitherto. In this study, we investigated the probiotic potential of novel pigmented bacterial strains isolated from the indigenous soil sediments in their vicinal habitats, which were screened for their antimicrobial activity against aquatic pathogens using agar well diffusion assay. The strains namely Exiguobacterium acetylicum (S01), Aeromonas veronii (V03), and Chryseobacterium joostei (V04) were phenotypically identified and confirmed by 16S rRNA gene sequence analysis. Further characterization revealed that strains S01 and V03 survive relatively in lower pH and higher bile salt concentrations and possess good adherence ability and broad-spectrum antibiotic susceptibility. The isolate S01 exhibited the higher adhesion ability to hydrocarbons (82%) and mannose-specific adhesion (msa) gene expression. Additionally, the probiotic effects were evaluated in Artemia nauplii fed with algae supplemented with S01, V03, and V04 strains (2.7 × 107 cfu/mL) for 3 days under axenic environment. We observed a significant increase (p < 0.05) in the survival rate of Artemia nauplii treated with S01 (83 ± 5%) and V03 (55 ± 5%), whereas the survival rate was only 30 ± 0% in the untreated group. Moreover, the individual length (IL) was increased in treated group S01 (156.7 ± 2.2 μm), V03 (146.1 ± 3.4 μm), and V04 (134.4 ± 2.5 μm) compared with untreated group (116.0 ± 4.8 μm). Our results revealed that E. acetylicum S01 exhibits desirable functional probiotic attributes compared to A. veronii and C. joostei and it would be a promising probiotic strain, which can be efficiently used in the aquaculture applications.
KeywordsAquaculture Pigmented bacteria Probiotics Artemia salina Gut microbiota
The authors wish to acknowledge DST-Science Engineering Research Board (SERB), Government of India, for the financial support under Young Scientist Scheme (SB/YS/LS-05/2014) and DST-PURSE Phase-II, M.KU and UGC-SAP of School of Biotechnology for instrumentation support. The authors wish to acknowledge Dr. B. S. Dileep Kumar, Senior Principal Scientist & Head, CSIR-National Institute for Interdisciplinary Science and Technology (NIIST), Thiruvananthapuram, for permitting to use SEM imaging facility. The authors also wish to acknowledge Mr. Abel Arul Nathan for his help in the statistical analysis.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that there is no conflict of interest.
The experiments in this study were performed in accordance with relevant national and international guidelines. This article does not contain any studies with human participants or animals performed by any of the authors.
- 2.Cordero H, Esteban MA, Cuesta A (2014) Use of probiotic bacteria against bacterial and viral infections in shellfish and fish aquaculture. INTECH Open Sci 8:239–265Google Scholar
- 6.Hill C, Guarner F, Reid G, Gibson GR, Merenstein DJ, Pot B, Morelli L, Canani RB, Flint HJ, Salminen S, Calder PC, Sanders ME (2014) Expert consensus document: the International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastroenterol Hepatol 11(8):506–514. https://doi.org/10.1038/nrgastro.2014.66 CrossRefGoogle Scholar
- 8.Medina CO, Cortes AL, Martinez AMM (2009) Aerobic Gram-positive heterotrophic bacteria Exiguobacterium mexicanum and Microbacterium sp. in the gut lumen of Artemia franciscana larvae under gnotobiotic conditions. Curr Sci 96:120–129Google Scholar
- 9.Gunasekara R, Rekecki A, Baruah K, Bossier P, Broeck WV (2010) Evaluation of probiotic effect of Aeromonas hydrophila on the development of the digestive tract of germ-free Artemia franciscana nauplii. J Exp Mar Biol Ecol 393(1-2):78–82. https://doi.org/10.1016/j.jembe.2010.07.006 CrossRefGoogle Scholar
- 11.Aliabadi MA, Ghasemi MF, Isaazadeh KH (2014) Antimicrobial activity bioactive compounds produced by Exiguobacterium acetylicum PTCC1756 against pathogenic bacteria. Sci J Microbiol 3:55–62Google Scholar
- 17.Marques A, Dinh T, Ioakeimidis C, Huys G, Swings J, Verstraete W, Dhont J, Sorgeloos P, Bossier P (2005) Effects of bacteria on Artemia franciscana cultured in different gnotobiotic environments. Appl Environ Microbiol 71(8):4307–4317. https://doi.org/10.1128/AEM.71.8.4307-4317.2005 CrossRefGoogle Scholar
- 18.Cintas LM, Rodriguez JM, Fernandez MF, Sletten K, Nes IF, Hernandez PE, Holo H (1995) Isolation and characterization of pediocin L50, a new bacteriocin from Pediococcus acidilactici with a broad inhibitory spectrum. Appl Environ Microbiol 61(7):2643–2648Google Scholar
- 19.Vos P, Garrity G, Jones D, Krieg NR, Ludwig W, Rainey FA, Schleifer KH, Whitman WB (eds) (2009) Bergey’s manual of systematic bacteriology, vol 3, 2nd edn. Springer-Verlag, New YorkGoogle Scholar
- 20.Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucl Acids Res 22(22):4673–4680. https://doi.org/10.1093/nar/22.22.4673 CrossRefGoogle Scholar
- 21.Saitou N, Nei M (1987) The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4(4):406–425Google Scholar
- 23.CLSI (2011) Clinical and Laboratory Standards Institute (CLSI): Performance standards for antimicrobial susceptibility testing: twenty-first informational supplement M100–S21. Wayne, PA, USA: 30:65Google Scholar
- 25.Sorgeloos P, Lavens P, Leger P, Tackaert W, Versichele D (1986) Manual for the Culture and use of brine shrimp Artemia in aquaculture. Artemia Reference Center, Faculty of Agriculture, State University of Ghent, Belgium, p 318Google Scholar
- 27.Cruz PM, Ibanez AL, Monroy Hermosillo OA, Ramirez Saad HC (2012) Use of probiotics in aquaculture. Int Scholarly Res Network Microbiol 13:916845Google Scholar
- 28.Maeda M, Liao IC (1994) Microbial processes in aquaculture environment and their importance for increasing crustacean production. Japan Int Res Center Agric Sci 28:283–288Google Scholar
- 29.Garriques D, Arevalo G (1995) An evaluation of the production and use of a live bacterial isolate to manipulate the microbial flora in the commercial production of Penaeus vannamei postlarvae in Ecuador, 53–59. In: Browdy CL, Hopkins JS (ed) Swimming through troubled water. Proceedings of the Special Session on Shrimp Farming, Aquaculture 95. World Aquaculture Society Baton Rouge, LaGoogle Scholar
- 31.Powedchagun P, Suzuki H, Rengpipat S (2011) Characterization of a probiotic Bacillus S11 bacterium of black tiger shrimp Penaeus monodon. Songklanakarin J Sci Technol 33:1–8Google Scholar
- 35.Giri SS, Sukumaran V, Dangi NK (2012) Characteristics of bacterial isolates from the gut of freshwater fish, Labeo rohita that may be useful as potential probiotic bacteria. Pro Antimicrobial Prot 4:238e242Google Scholar
- 38.Mohapatra S, Chakraborty T, Prusty AK, Das P, Paniprasad K, Mohanta KN (2012) Use different microbial probiotic in the diet of rohu, Labeorohita fingerlings: effect on growth, nutrient digestibility and retention, digestive enzyme activities and intestinal microflora. Aquacult Nutr 18(1):1–11. https://doi.org/10.1111/j.1365-2095.2011.00866.x CrossRefGoogle Scholar
- 39.Hosoi T, Kiuchi K (2003) Handbook of fermented functional foods. CRC Press 227–245Google Scholar
- 42.Bharathi K, Akila M, Selvakumar D (2016) Evaluation of probiotic characterization of marine bacteria and its growth performance in zebrafish. Imper J Interdisci Res 2:2129–2137Google Scholar
- 43.Schrehardt A (1987) Ultrastructural investigations of the filter-feeding apparatus and the alimentary canal of Artemia. In: Sorgeloos P, Bengtson DA, Decleir W, Jaspers E (eds) Artemia research and its applications. Universa Press, WetterenGoogle Scholar