![The following parameters were calculated per group over 40 days of feeding trial (until fish have
quadruplicated in weight):
Feed intake during the trial
Specific growth rate (SGR): 100 x [(ln final body weight – ln initial body weight)/days]
Feed conversion ratio (FCR): Feed intake/weight gain
Fish Survival (%)
Feeding trials and growth performance
No differences in Growth Performance were found betwen control and test
groups
Ljubljana, 25 settembre 2019](https://image.slidesharecdn.com/lubiana2parati25september2019-191030113932/85/The-Microalgae-hub-project-SaltGae-Final-event-17-320.jpg)
The Microalgae hub project - SaltGae Final event
- 1. «THE MICROALGAE HUB» Microalgae for the treatment and valorization of agro-zootechnical waste and dairy by-products How to use by-products from the food industry to grow microalgae and their valorization SaltGae, final meeting Ljubljana , 25 September 2019 Katia Parati Istituto Spallanzani
- 2. Dairy by-products characterization Campioni N-NH4 (mg/L) P-PO4 (mg/L) TKN (mg/ L) COD tot (g/L) COD sol (g/L) ST (g/kg) SV (g/kg) N/P COD/SV SV/ST Whey 65 455 133 6 87,3 65,6 64,2 59,22 2,9 1,47 0,92 Exhausted whey 10 320 720 61,6 50,9 55,87 46,94 2,3 1,31 0,84 Buttermilk 7,7 130 223 15-20 5,9 14,53 12,84 1,7 1,60 0,88 Whey: very high COD, competition with bacteria Exhausted whey : very high COD, competition with bacteria Buttermilk: COD acceptable, N / P unbalanced vs P, N need to be hydrolyzed, competition with bacteria Rivolta d’Adda, 16 settembre 2019 2,5 milion of tons/year in Lombardy region
- 3. Experimental Design • Chlorella sorokiniana • Scenedesmus acuminatus • Acutodesmus obliquus • Desmodesmus armatus • Chlamydomonas debaryana • Porphyridium cruentum • Euglena gracilis • Arthrospira platensis ("Spirulina") Exhausted whey Laboratory evaluation of microalgal species able of growing on dairy by-products Buttermilk Ljubljana, 25 settembre 2019
- 4. Arthrospira platensis (Spirulina) Generelly recognized as safe (GRAS) since 1981 (FDA) • Antioxidant • Anti-inflammatory • hypoglycemic • antihypertensive • immunostimulant In 1967 Spirulina was recognized as a ‘future food source’ by the International Association of Applied Microbiology Ljubljana, 25 settembre 2019
- 5. Valorisation of Arthrospira platensis in the cosmetic and nutraceutical sector PHYCOCYANIN: association of proteins of the phycobiliprotein family, and of water-soluble photosynthesis pigments, the phycocynobilins, of the phycobiline family. • powerful antioxidant (40 times higher than that of vitamin C and vitamin E). Contrast oxidative stress. • it counteracts inflammatory states by acting similarly to non-steroidal anti-inflammatory drugs (Fans) and by inhibiting various enzymes involved in the inflammatory response (colitis, inflammation to the ear, arthritis and tumors). • it contributes to the reinvigoration of the immune system by strengthening the cell membrane and increasing the protection of cells from virus attacks. • alleviate the symptoms of seasonal allergies. • …….. Ljubljana, 25 settembre 2019
- 6. Ljubljana, 25 settembre 2019 26 ottobre 2018 CLINICAL TRIAL ClinicalTrials.gov (last access 10/09/2019) “Spirulina” -23 "completed" studies in the world -17 studies in the world "being completed»
- 7. Valorisation of Arthrospira in the cosmetic sector Wang et al. Bioresource Technology 2015 184:355–362 Ljubljana, 25 settembre 2019
- 8. Spirulina in animal feed • It induces increasing in fat, protein and lactose content in cow's milk (Simkus et al., 2007- 2008), whose production can increase by 21% (Kulpys et al., 2009). • Improves growth and fertility (Holman, 2013) • In common carp it improves growth and increases proteins and fat content (Abdulrahman and Ameen, 2014) • It stimulates pigmentation in ornamental fish, such as Koi carp (Gouveia, 2003) • It stimulates the immune system of trout juveniles increasing resistance to pathogens (Yeganeh, 2015). Ljubljana, 25 settembre 2019
- 9. What’s the aim of the work? • decrease the cost of the biomass production by reducing the cost of synthetic medium; • valorize the dairy by-product, reducing at the same time the nutrient content (N, P, COD) Ljubljana, 25 settembre 2019
- 10. Arthrospira platensis (spirulina) cultivation test on exhausted whey: pilot scale test (semi-batch mode) Cultivation of A. platensis for nutrients removal from dairy by-products Parameters Characteristics exhausted whey Characteritics effluent pH 5,5 9 COD (g/L) 50 0,5 N-NO3 (mg/L) <1 0 N-NH4 (mg/L) 10 0 N-tot (mg/L) 720 50-100 Ntot removal : 76% COD Removal: 94% Constant growth Ljubljana, 25 settembre 2019
- 11. Harvesting and conservation Filtration lyophilization Ljubljana, 25 settembre 2019
- 12. g/100g Humidity (%) Protein (%) Carbohydrates (%) Lipids (%) Arthrospira platensis (control) 8,5 51.8 10.3 12,5 Arthrospira platensis (hexausted whey) 11,3 51.1 11,2 10,5 Biochemical and microbiological analysis Samples Total microbial charge Enterobateriacae E. coli sporogenous sulfite reducers FA Salmonella Arthrospira platensis (control) 9,5E+04 <100 <100 <100 - assente Arthrospira platensis (hexausted whey) 1,2E+05 <100 <100 <100 - assente Data expressed as ufc/g CREA_ZA Ljubljana, 25 settembre 2019
- 13. Control – pilot scale test– pilot scale Fatty acids composition of Spirulina (test vs control) Monounsaturated polyunsaturatedsaturated Test: Spirulina grown on synthetic medium Control: Spirulina grown on exhausted whey Ljubljana, 25 settembre 2019
- 14. - Freshwater fish of zootechnical interest, especially for caviar production. Caviar market, world production: 250 ton / year (over € 400 million per year), with great growth potential. Italy is the European leader for caviar production (50 tonnes in 2018) and second worldwide (between China and Russia) - Sturgeons are mainly carnivorous and opportunistic omnivorous. At all stages, they spend most part of their life at the bottom of the river, to feed on algae attached to rocks, zooplankton, small invertebrates and fish. - The main problems of these fish, during the juvenile phase, are above all associated with the adaptation of new diets and, depending on the species, at this stage the mortality is often very high The sturgen: Acipenser baeri Feeding trial on Sturgeon juveniles (Acipenser baeri) VALORISATION OF ARTHROSPIRA IN THE FEED INDUSTRY Ljubljana, 25 settembre 2019
- 15. Feeding trial on Sturgeon juveniles (Acipenser baeri) Microalgae biomass production (from standard synthetic medium (MS) and HEXAUSTED WHEY External Collaborations: AIA-Veronesi (Verona), University of Almeria (Spain) and University of Milan Harvesting and drying Aquafeed preparation Feeding trail on sturgeon juveniles Ljubljana, 25 settembre 2019
- 16. Monofactorial random design diets tested in triplicate (control vs test) Control: groups of fish fed by control diet, based on fish meal; Test: groups of fish fed by experimental diet, based on 10% inclusion of spirulina. • Fish ~ 100 A. baerii (IBW 12,8 ± 0.4 g) randomly divided among 6 groups • Experimental facilities circular tanks (Volume 130 L) RAS T 18.9 ± 0.6°C, salinity 1± 0,02‰, DO 9.6 ±1.2mg/l, pH 8.1 ± 0.1, NH4-N <0.06mg/l, NO2-N <0.2mg/l, light–dark cycle 12:12 • Experimental protocol • Fish/tank: 16 • Feed ratio: 3% g/w • Feeding period: 6 weeks (6 days per week) • Diets offered in two daily meals • Fish were tank/group weighted every week Feeding trials and growth performance
- 17. The following parameters were calculated per group over 40 days of feeding trial (until fish have quadruplicated in weight): Feed intake during the trial Specific growth rate (SGR): 100 x [(ln final body weight – ln initial body weight)/days] Feed conversion ratio (FCR): Feed intake/weight gain Fish Survival (%) Feeding trials and growth performance No differences in Growth Performance were found betwen control and test groups Ljubljana, 25 settembre 2019
- 18. Valorization of Arthrospira platensis in cosmesis Cytotoxicity Test No negative effects of PHYCOCYANIN complex were found on human fibroblasts in the range of concentration tested Membrane Potential Testand ROS Evaluation Test Protection of the human fibroblasts against oxidant agent (H2O2: 1,5-3 mM) PRELIMINARY TRIAL Cytotoxicity and antioxidant activity assay on human fibroblasts, in order to test phyto-complex based on PHYCOCYANIN protein, extracted from Spirulina grown on a synthetic medium (Zarrouk) Ljubljana, 25 settembre 2019
- 19. Naviglio instrument: Solid / liquid extraction The generation of a negative pressure gradient between the outside and the inside of a solid matrix containing extractable material, followed by a sudden restoration of the initial equilibrium conditions, induces the forced extraction of the compounds in the solvent. Phycocyanin complex Extraction Spirulina Valorization of Arthrospira platensis in cosmesis Ljubljana, 25 settembre 2019
- 20. and aquafeed industry UNDER STUDIES : -Extraction of phycocyanin from Spirulina grown on exhausted whey; -Cytotoxicity and antioxidant power test on fibroblasts -Comparison of the results with those obtained using phycocyanin complex grown on synthetic medium (Zarrouk) Valorization of Arthrospira platensis in cosmesis Ljubljana, 25 settembre 2019
- 21. Thanks for your attention! Katia ParatiElena Ficara Valeria Mezzanotte Aldo Tava Barbara Pisaroni Donatella Melani Ilaria Casadei Federico Castillo Luciano Foglio Lorenzo Proietti Tiziana Bongiorno Katia Parati