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Effect of dietary probiotic supplementation on growth and innate immune system in sole (Solea senegalensis Kaup, 1858) subjected to pathogenic bacteria infection and handling stress

Andreia F. Domingues1, 2, Maria João D. Peixoto1, Sónia M. G. Batista1, 3, Ana Gomes4, Célia M. Manaia4, José F. M. Gonçalves1, 3, Sofia Cunha4, Rui Barros5, Luísa M. P. Valente1, 3, Benjaním Costas1 and Rodrigo O. A. Ozorio1, 3*
  • 1 CIIMAR – Interdisciplinary Centre of Marine and Environmental Research., Portugal
  • 2 ESTM – School of Tourism and Maritime Technology – Polytechnic Institute of Leiria, Portugal
  • 3 ICBAS – Abel Salazar Institute for Biomedical Sciences – University of Porto, Portugal
  • 4 CBQF – Centre of Biotechnology and Fine Chemistry– Faculty of Biotechnology of the Catholic University of Portugal (ESB)., Portugal
  • 5 Biostrument - Development Consulting Projects Biochemicals S.A., Portugal

Stress could change disease susceptibility by affecting immune function and disease resistance in fish. Therefore disease is not just the presence of pathogens but an outcome from a combination of factors. Photobacterium damselae subsp. piscicida is a pathogenic bacterium that causes great economic losses in the sole aquaculture. Probiotics could be a good alternative for the control of some bacterial diseases preventing in this way the use of antibiotics. The objectives of this study were to evaluate the effects of probiotic supplementation on growth, innate immune response and protection against Photobacterium damselae subsp. piscicida in Senegalese sole (Solea senegalensis) juveniles.

Fish were fed on six experimental diets, one non-supplemented probiotic diet (CTL), two mono-species probiotic diets (PB1–Shewanella spp and PB2–Enterococcus spp) and three multi-species probiotic diets (PB3–Shewanella spp and Arthrobacter spp, PB4–Pseudomonas spp and Arthrobacter spp, PB5–Shewanella spp, Arthrobacter spp and Enterococcus spp). Fish were kept in a semi-closed recirculating sea-water system (19°C, 30‰) and fed ad libitum the dietary treatments 3 times per day for 12 weeks. Growth performance and innate immune parameters (peroxidase and lysozyme activities and alternative complement pathway - ACH50) were analyzed.

After the growth trial, two infection trials (trial A and B) were carried out for 15 days to test the interaction effect between two variables: pathogenic infection (Photobacterium damselae subsp. piscicida) and stress associated with handling (transport). Trial A: animals were infected on the same day of transport; trial B: animals were transported and acclimatized for 7 days before infection. Stimulation Index (SI) of peroxidase activity was only determined for the infection trials, and according Díaz-Rosales (2009). SI is determined by dividing the value of each probiotic treatment by the average of the non-supplemented probiotic diet (CTL). Values higher than 1 reflect an SI increase and lower than 1 a decrease, in relation to the dietary CTL group.

Fish fed PB2 (22.91±0.53g) and PB4 (24.03±0.48g) diets showed less weight gain compared to fish fed CTL (26.56±0.63g) diet. Probiotics diets did not confer significant differences in innate immune parameters of sole juveniles compared to the control diet. However PB5 probiotic supplementation, presenting lower peroxidase activity (38.68±10.82 EU mL-1) compared to CTL group (90.30±22.99 EU mL-1).

After both infection trials, the cumulative mortality was not significantly different, regardless the dietary treatment and the stress factors tested. Generally, trial A (0.91±0.75) tended to have higher SI peroxidase activity than trial B (0.27±0.22). For trial A, SI was significantly lower in fish fed PB1 (0.31±0.39) than in fish fed PB4 (1.31±0.59) and PB5 (1.12±0.66). Moreover, fish fed PB3 (0.36±0.39) diet had significantly lower SI values than PB4 (1.31±0.59).

Our data suggest that the innate immune responses of sole was not influenced by diet alone, but it shows some changes in combination with stress associated with handling.

Acknowledgements

S. M. G. Batista and B. Costas are supported by FCT – SFRH/BD/76668/2011 and SFRH/BPD/77210/2011, respectively. This work was also supported by PROBIOSOLEA project with the financial support of Quadro de Referência Estratégico Nacional – QREN and Programa Operacional Regional do Norte – ON2 (Ref. no. 13551), supported by the European fund for regional development FEDER.

References

Díaz-Rosales, P., Arijo, S., Chabrillón, M., Alarcón, F.J., Tapia-Paniagua, S.T., Martínez-Manzanares, E., Balebona, M.C. & Moriñigo, M.A. (2009) Effects of two closely related probiotics on respiratory burst activity of Senegalese sole (Solea senegalensis, Kaup) phagocytes, and protection against Photobacterium damselae subsp. piscicida. Aquaculture, 293, 16-21.

Keywords: Growth, Immune System, Photobacterium damselae subsp. piscicida, Probiotics, Solea senegalenses

Conference: IMMR | International Meeting on Marine Research 2014, Peniche, Portugal, 10 Jul - 11 Jul, 2014.

Presentation Type: Poster Presentation

Topic: AQUACULTURE

Citation: Domingues AF, Peixoto MD, Batista SG, Gomes A, Manaia CM, Gonçalves JM, Cunha S, Barros R, Valente LP, Costas B and Ozorio RA (2014). Effect of dietary probiotic supplementation on growth and innate immune system in sole (Solea senegalensis Kaup, 1858) subjected to pathogenic bacteria infection and handling stress. Front. Mar. Sci. Conference Abstract: IMMR | International Meeting on Marine Research 2014. doi: 10.3389/conf.fmars.2014.02.00019

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Received: 13 May 2014; Published Online: 18 Jul 2014.

* Correspondence: Dr. Rodrigo O. A Ozorio, CIIMAR – Interdisciplinary Centre of Marine and Environmental Research., Porto, Portugal, rodrigo.ozorio@ciimar.up.pt