24h storage in starvation • Pathogen loads are believed to be an important factor in fish larvae survival due to their limited immune system; • Larvae mortality is often caused by opportunistic bacteria introduced with the life feed supplied (Planas and Cunha, 1999); • Artemia sp. is widely used as live food in larviculture, and a known vector for introducing bacteria into the systems (Makridis et al., 2010); • Artemia low temperature storage is a methodology to preserve size and nutritional profile with no scientific data on bacteria proliferation. F. Soares*, S. Castanho, M. Moreira, A.C. Mendes and P. Pousão-Ferreira Aquaculture Research Station, National Institute for the Ocean and Atmosphere (IPMA), Av. 5 de Outubro, s/n 8700-305 Olhão, Portugal ü Cold storage (5±1ºC) seems to be also a viable methodology to restrain total bacterial proliferation after the enrichment process, temperature affected the amount of total bacteria but not their diversity. ü Bacteria diversity present in the enriched Artemia seems to be related with the enrichment products used, rather than with the temperature. • Buller N.B. 2004. Bacteria from fish and other aquatic animals: a practical identification manual, 1st Edition, CABI Publishing, pp.171-176; • Holt J.G. 1994. Bergey's Manual of Determinative Bacteriology, 9th Edition, Williams & Wilkins.; • Høj L., Bourne D.G., Hall M.R. 2009. Localization, abundance and community structure of bacteria associated with Artemia: Effects of nauplii enrichment and antimicrobial treatment. Aquaculture, 293: 278–285; • Kellam S.J. and J.M. Walker. 1989. Antibacterial activity from marine microalgae in laboratory culture. British Phycological Journal 24:191-194; • Makridis P., L. Ribeiro, M.T. Dinis. 2010. Influence of microalgae supernatant, and bacteria isolated from microalgae cultures, on microbiology, and digestive capacity of gilthead seabream (Sparus aurata) and senegalese sole (Solea senegalensis) larvae. Journal of the World Aquaculture Society 41(5):780-790; • López-Torres M.A., Lizárraga-Partida M.L. 2001. Bacteria isolated on TCBS media associated with hatched Artemia commercial brands. Aquaculture, 194:11-20; • Planas M. and I. Cunha. 1999. Larviculture of marine fish: problems and perspectives. Aquaculture 177:171-190; • Snoussi M., K. Chaieb, R. Mahmoud, and R. Bakhrouf. 2006. Quantitative study, identification and antibiotics sensitivity of some Vibrionaceae associated to a marine hatchery, Annals of Microbiology 56(4):289-293; • Srinivasakumar K.P. and M. Rajashekhar. 2009. In vitro studies on bactericidal activity and sensitivity pattern of isolated marine microalgae against selective human bacterial pathogens. Indian Journal of Science and Technology 2(8):16-23. INTRODUCTION BIBLIOGRAPHY MATERIALS AND METHODS * [email protected]CONCLUSIONS ACKNOWLEDGMENTS DISCUSSION ü Artemia enriched with GD showed a lower proportion of Vibrionaceae possibly due to an effect of antimicrobial molecules produced by the microalgae (Kellam and Walker, 1989; Srinivasakumar and Rajashekhar, 2009); ü The Vibrionaceae identified are similar to the species found by López- Torres & Lizárraga-Partida (2000) and Høj et al. (2009), when they´ve analysed the bacterial community of Artemia; ü Vibrio alginolyticus was common in the two enrichments, which is in agreement with Snoussi et al. (2006), that reports this bacteria as the most common bacteria in Artemia. Characterization of the bacteria flora associated with Artemia sp. after enrichment and stored at two different temperatures. Microbial characterization of enriched Artemia sp. at two different temperatures and enrichments GD - GREEN DIETS, (produced by Necton S.A.) experimental emulsion of several freeze-dried microalgaes supplemented with DHA OBJECTIVE Enrichment RP - Red Pepper®, Bernaqua™ commercial product Trial setup: Analytical methodology: RESULTS Fig. 1. Total bacteria and Vibrionacea present in the different treatments.GD- enriched microalgae, RP- Red Pepper®,i- initial sample, (-)5ºC, (+) 19ºC. Letters a, b, c and d indicate different groups with statistical significant differences (Wilcoxon Rank test, P<0.05). Bacterial quantification Vibrionaceae identification GD - Green Diets RP - Red Pepper Other bacteria ü Moritella marina / Vibrio aestuarians ü Vibrio coralliilyticus ü Vibrio vulnificus ü Vibrio pelagius biovar . 2 Common bacteria Vibrio alginolyticus Room with controlled temperature GD+ 19±1ºC common temperature for marine fish larvae rearing GD- 5±1ºC RP+ 19±1ºC common temperature for marine fish larvae rearing RP- 5±1ºC Room with controlled temperature Cooling tank Cooling tank Samples were collected: 1. After enrichment (GDi and RPi) 2. After 24h storage (GD+; GD-; RP+ and RP-) 10ml Artemia + Water Homogeneization Plating TSA TCBS CFU´s counting (7 days) Selection and isolation of bacteria Biochemical identification of bacteria (Holt, 1994; Buller, 2004) Florbela Soares acknowledge Ciência 2008 program (FCT)
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Microbial characterization of enriched Artemia sp. at two ... · Artemia sp. is widely used as live food in larviculture to feed marine fish and penaeid shrimps and a known vector
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24h storage in starvation
• Pathogen loads are believed to be an important factor in fish larvae
survival due to their limited immune system;
• Larvae mortality is often caused by opportunistic bacteria introduced with
the life feed supplied (Planas and Cunha, 1999);
• Artemia sp. is widely used as live food in larviculture, and a known vector
for introducing bacteria into the systems (Makridis et al., 2010);
• Artemia low temperature storage is a methodology to preserve size and
nutritional profile with no scientific data on bacteria proliferation.
F. Soares*, S. Castanho, M. Moreira, A.C. Mendes and P. Pousão-Ferreira Aquaculture Research Station, National Institute for the Ocean and Atmosphere (IPMA), Av. 5 de Outubro, s/n 8700-305 Olhão, Portugal
ü Cold storage (5±1ºC) seems to be also a viable methodology to restrain total bacterial proliferation after the enrichment
process, temperature affected the amount of total bacteria but not their diversity.
ü Bacteria diversity present in the enriched Artemia seems to be related with the enrichment products used, rather than with
the temperature.
• Buller N.B. 2004. Bacteria from fish and other aquatic animals: a practical identification manual, 1st Edition, CABI Publishing, pp.171-176; • Holt J.G. 1994. Bergey's Manual of Determinative Bacteriology, 9th Edition, Williams & Wilkins.; • Høj L., Bourne D.G., Hall M.R. 2009. Localization, abundance and community structure of bacteria associated with Artemia: Effects of nauplii enrichment and antimicrobial treatment. Aquaculture, 293: 278–285; • Kellam S.J. and J.M. Walker. 1989. Antibacterial activity from marine microalgae in laboratory culture. British Phycological Journal 24:191-194;• Makridis P., L. Ribeiro, M.T. Dinis. 2010. Influence of microalgae supernatant, and bacteria isolated from microalgae cultures, on microbiology, and digestive capacity of gilthead seabream (Sparus aurata) and senegalese sole (Solea
senegalensis) larvae. Journal of the World Aquaculture Society 41(5):780-790; • López-Torres M.A., Lizárraga-Partida M.L. 2001. Bacteria isolated on TCBS media associated with hatched Artemia commercial brands. Aquaculture, 194:11-20;• Planas M. and I. Cunha. 1999. Larviculture of marine fish: problems and perspectives. Aquaculture 177:171-190; • Snoussi M., K. Chaieb, R. Mahmoud, and R. Bakhrouf. 2006. Quantitative study, identification and antibiotics sensitivity of some Vibrionaceae associated to a marine hatchery, Annals of Microbiology 56(4):289-293; • Srinivasakumar K.P. and M. Rajashekhar. 2009. In vitro studies on bactericidal activity and sensitivity pattern of isolated marine microalgae against selective human bacterial pathogens. Indian Journal of Science and Technology 2(8):16-23.