Bandarra, Narcisa

Castro, Paula M. L.

Morais, R.

Oliveira, Susana

Raposo, M. Filomena de J.

English

Repositório Institucional da Universidade Católica Portuguesa

MATERIAL AND METHODOLOGIESTable 1. Characteristics of a brewery effluent: range of values for indicated parameters.ON THE UTILIZATION OF THE ALGAL BIOMASS GROWN IN A BREWERY EFFLUENT IN ANIMAL FEEDM Filomena de J-Raposo1, Susana Oliveira1, Paula M L Castro1, Narcisa Bandarra2 and R Morais11- Escola Superior de Biotecnologia - Universidade Católica Portuguesa Rua Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal2- Instituto de Investigação das Pescas e do Mar Av. Brasília, 1449-006 LisboaCharacteristics of the effluent Range of values CBO5 560-4778mg O2/L CQO 565-7837mg O2/L Ammonium 0.173-5.913mM Nitrate 0.030-0.180mM Phosphate 0.597-3.430mM  INTRODUCTIONTreatment of effluents and wastewaters from the Agro-FoodIndustries has a relevant importance, both in Europe and inPortugal, either at the economical level but also from theenvironmental point of view.One of the most profitable biological methods could be microalgaeutilization. Besides, after treatment, there can be some algalbiomass valorization in animal feed, as a protein source, or usingsome of their components like pigments, enzymes.In this study, besides the evaluation of a brewery effluentutilization as a growing medium for microalgae, it was also shownthat in the end, the algal biomass could be used as animal feed,after being analysed for protein, fatty acid content and heavymetals, eventually incorporated.RESULTS AND DISCUSSIONTable 2. Effective capacity of treatment of the brewery effluent by the microalga Chlorella vulgaris and by themicroalgae consortium obtained from the autochthonous flora of the same effluent.In what concerns the quality of the protein, a significant increase in the aminoacids contentwas observed. Besides, from the analysis to the Chlorella vulgaris biomass, it was verifiedthat aminoacids concentration was always higher in the algae grown with effluent,difference being statistically significant both at essential and total aminoacids. Moreover,the highest results were obtained with the biomass that grew in the effluent diluted to 1:1(Figure 1). However, the ratio essential aminoacids/total aminoacids is not different in allthe conditions.Figure 1. Growth curves of C. vulgaris (on the left) and the autochthonous flora (on the right) cultures, ineffluent as it is (Effl) and diluted, against a control, in the usual growth medium.Figure 2. Different percentages obtained for fatty acid composition (on the left) and aminoacids content (on theright) with the biomass grown in the effluent as the growth medium, against the control (usual growthmedium).Although fatty acids content showed a general decrease with the increase of the effluentlevel, they also present an increase in the ramified chain fatty acids (figure 2).Figure 3. Contents in mercury, cadmium, nickel, and leadin the biomass samples from the effluent, either as it is ordiluted, and the control (usual culture medium).REFERENCES↦ (1) Jeffrey, S. W. e Humphrey, G. F. (1975). New Spectrophotometric Equations for Determining Chlorophylls a, b, c1 and c2 in Higher Plants, Algae and Natural Phytoplankton. Biochem. Physiol. Pflanzen (BPP),Bd 167, S. 191-194.↦ (2) Standard Methods for the Examination of Water and Wastewater (1998). Clescers, L S, Eaton, A D, Greenberg, A E (Ed.), 20th edition. APHA, AWWA, WEF.↦ (3) Weatherburn, M. W. (1967). Phenolhypochlorite reaction for determination of ammonia. Anal. Chem., 39: 971.↦ (4) AOAC (1998). Official Methods of Analysis, 16th ed., 4th revision, Vol. I e II. Association of Official Analytical Chemistry, AOAC International, Gaithersburg MD.↦ (5) Lepage, G. e Roy C. C. (1986). Direct transesterification of all classes of lipids in a one-step reaction. J. Lipid Res., 27: 114-119.↦ (6) Cohen, Z.; Vonshak, A. and Richmond, A. (1988). Effect of environmental conditions on fatty acid composition of the red alga Porphyridium cruentum: correlation to growth rate. J. of Phycol, 24: 328-332.↦ (7) AOAC (1990). Official Methods of Analysis, 15th ed. Vol. 1. Association of Official Analytical Chemistry, AOAC International, Arlington.↦ (8) NP 2928 (1988). Determinação do Teor de Mercúrio. Método espectrofotométrico de absorção atómica. Instituto Português da Qualidade, Lisboa, 5p.↦ (9) Directiva Comunitária 1999/29/EC (1999). Substâncias e produtos indesejáveis na nutrição animal.036912150 2 4 6 8 10 12 14 16Time (days)Total chlorophyll (mg/L)Control1:21:1Efl.as it  is0204060800 2 4 6 8 10 12 14 16 18 20Time (days)Total chlorophyll (mg/L)Control1:21:1Efl.as it  isACKNOWLEDGEMENTSThis work was supported by Project “Novos Sistemas de Reciclagem de Efluentes Agro-Industriais usando Biotecnologia de Microalgas com Valorização da Biomassa”, from ProgrammePraxis XXI. The authors would like to thank UNICER-União Cervejeira SA for the effluent, andalso Rui Mendes and Pedro Castro for their helpful work.Evaluated parametersInitial values Percentageof removalInitial values Percentageof removalChlorella vulgaris Microalgae consortiumCBO5 560-2730 1340-2354 18.3-27.1CQO 800-3900 2172-3846 12.6-14.6nitrogen (mM) C             0.404 -0.585Effluent 5.302-10.26130.9-59.584.7-98.0C             3.563 -8.251Effluent 7.935-10.50239.2-62.831.3-98.0phosphate (mM) C             0.144 - 0.429Effluent 0.597-3.43066.411.8-53.9C            0.116 - 0.223Effluent 0.109-0.99337.8-85.210.1-93.0In what concerns the incorporation ofmetals, concentration in the algalbiomass is as high as the concentration inthe growth medium. The results (figure3) are in the maximum range valuesestablished by the EC, in animal feed (9).Ni is the only determined metal whosemaximum admissible limits are not yetlegislated; it presents, nevertheless, alower toxicity than the other heavymetals.0,0 2,0 4,0 6,0 8,0 10,0Control 1:2 1:1Effl as it isFatty Acids (g/100g)14:0isobr16:117:118:2w618:3w318:4w320:5w30,0 3,0 6,0 9,0 12,0 15,0 18,0Control 1:2 1:1Effl as it isAminoacids (g/100g)PhenylalanineIsoleucineLeucineLysineMethionineThreonineValine0,0 2,0 4,0 6,0 8,0 10,0 12,0 14,0Control 1:2 1:1Eff as it isHeavy Metals (ppm)MercuryCadmiumNickelLeadIn this work, not only Chlorella vulgaris but alsothe autochthonous flora from the residual waterhave shown a increase in their growth, moreevident when a effluent from a brewery wasused as the culture medium (Figure 1).These results are supported by previous works,when high rates of N and P removal wereobserved along with a more evident growth ofthe algae. Besides, it was also observed that, ingeneral, rate of N removal was more efficient that P removal (Table 2). in 80% acetone, and determined byJeffrey and Humphrey equations (1).Growth curves for biomass evaluation:Characteristics of the effluent (table 1): Analyses to the biomass: BOD (2) COD (2) Phosphates Vanadomolibdophosphoric Acid Colorimetric Method (2)Ammonium Phenate Methodology (3) Nitrates Merck (Spectroquant) kit and OD read on a UV spectrophotometer, at 338nm. Aminoacids (4)  Fatty Acids (5) (6) Pb (7) Cd (7) Ni (7) Hg (8)Statistical analyses:Effluent:Experiment:Analyses: Nitrogen (autochthonous flora)corrected to 9.5mM (as nitrate)  Phosphorus (autochthonous flora)corrected to 0.124mM  triplicates duplicatescultures grown in plastic bag reactors as it is diluted with distilled water in triplicatesANOVA  Student’s t-test Non parametric testsKruskal and WallisMann-WhitneyMicroalgae:Chlorella vulgarisautochthonous flora from the effluentdirect counting (C. vulgaris)chlorophyll quantification (autochthonous flora)

On the utilization of the algal biomass grown in a brewery effluent in animal feed

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On the utilization of the algal biomass grown in a brewery effluent in animal feed

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