Thesis (MSc)--University of Stellenbosch, 2008. / Bibliography. Also available via the Internet.

Tesis de grado presentado para optar el titulo de Licenciature en Biologia, Universidad Mayor De San Simon, Facultad de Ciencias y Technologia, Carrea Biologia. / Abstract in Spanish and English.

Master of Science / Food Science Institute - Animal Science & Industry / Fadi M. Aramouni, Bean, Scott R. / The biochemical, physical and baking properties of caroubin, the main protein in the carob bean, were characterized. The biochemical properties of caroubin were analyzed using reversed-phase high performance liquid chromatography (RP-HPLC), size exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALS) and micro-fluidics analysis. The physical and baking properties of caroubin were characterized via SE-HPLC, laser scanning confocal microscopy, farinograph mixing, and texture profile analyzer analysis. Using a modified Osborne fractionation method, carob germ flour proteins were found to contain ~32% albumin and globulin and ~68% glutelin with no prolamins detected. When divided into soluble and insoluble protein fractions under non reducing conditions it was found that caroubin contained (~95%) soluble proteins and only (~5%) insoluble proteins. As in wheat, SEC-MALS analysis showed that the insoluble proteins had a greater Mw than the soluble proteins and ranged up to 8x107 Da. These polymeric proteins appeared to play a critical role in protein network formation. Analysis of the physical properties of carob germ protein-maize starch dough showed that the dough’s functionality was dependent on disulfide bonded protein networks, similar to what is found in wheat gluten. When baked into a bread these proteins were shown to have a possible improving affect by decreasing staling in gluten-free breads. This was evident when compared to a gluten-free batter bread, and a wheat bread over a five day period.

Thesis (Msc Food Sc (Food Science))--Stellenbosch University, 2008. / Carob (Ceratonia silliqua) is an evergreen, drought resistant tree of Mediterranean origin. Popularly known as St John’s bread, the carob pod has a long history of use in food (over 4 000 years). Carob has a good nutritional value, a long shelf-life (2-3 years) and it is relatively cheap. Due to its high sugar content, carob is naturally sweet. It also has a nutty chocolate-like flavour, but unlike chocolate or cocoa, carob does not contain any caffeine, thiobromine or oxalic acid. In addition, carob is normally regarded as a healthy food because of its low fat content (0.2 – 2.3%). Carob trees are also found in South Africa, especially in the Western Cape Province. Locally, carob trees have been used mainly ornamentally or as a source of animal fodder, with minimal use of the pods as a nutritious food source. Knowledge of the nutritional composition and the overall nutritional potential of locally (South African) grown carob cultivars is also limited. Carob could potentially be used as an alternative food source in South Africa as currently, most of this nutritious product goes to waste each year. In this study, the feasibility of using carob pods as an alternative source of food in South Africa was investigated. This was done by firstly, analysing the cultivars for proximate composition (moisture, carbohydrates, sugars, dietary fibre, protein, polyphenols, fat and ash) as well as for amino acids, fatty acids and minerals, in order to determine and compare their nutritional contents. Five cultivars (Tylliria, SFax, Aaronsohn, Santa Fe and an “Unknown” cultivar) were examined. The average proximate composition of raw carob pods was 8.17 – 9.56% moisture, 89.57 – 91.12% carbohydrates, 40.69 – 54.74% total sugars (33.70 – 45.09% sucrose, 1.79 – 4.95% glucose and 1.80 – 5.19% fructose), 29.88 – 36.07% dietary fibre, 3.07 – 4.42% protein, 2.58 – 3.08% polyphenols, 0.45 – 0.86% fat and 2.13 – 2.69% ash. Seven essential amino acids were present in all the cultivars, except for methionine which was not detected in the Single unknown cultivar. This study has shown that all the cultivars had good long-chain fatty acid (LCFA) proportions in terms of the saturated to polyunsaturated fatty acid (SFA: PUFA) and n-6 to n-3 ratios. The short-chain fatty acid content of the cultivars was low. All nine minerals (calcium, phosphorus, potassium, magnesium, sodium, manganese, iron, copper and zinc) analysed for in this study were detected in all five carob cultivars and all cultivars were very low in sodium. The impact of various roasting times (45, 60 and 75 min) at 150ºC, on the temperature sensitive components such as sugars, protein and fat, was also examined. Roasting had no significant (P>0.05) effect on the fat content. Although roasting significantly (P<0.05) reduced the sugar and protein content from 54.74 to 32.53% and 3.59 to 3.18%, respectively, levels in both raw and roasted carob still represented a potentially nutritious food source and alternative to cocoa. A variety of food products targeted at the various food market sectors were developed with carob as an ingredient. The formulations for five new food products (bread, porridge, breakfast cereal, mousse and milk-based drink) were developed where carob had successfully been incorporated as an ingredient. Microbiological and consumer sensory analyses carried out showed that all products developed were safe and acceptable. The findings of this study provide useful scientific evidence towards the fact that carob could potentially be used as an alternative food source in South Africa.

Master of Science / Department of Food Science / Fadi Aramouni / Despite the expansion of the gluten-free (GF) food market, some GF food items are still characterized by an overall mediocre quality. The effects of different types of egg ingredients (fresh, whites, dried) and carob germ flour (CGF) as well as par-baking technology on the quality of dough-based gluten-free sorghum dinner rolls were evaluated. Gluten-free rolls containing 30% of fresh shell eggs or equivalent of egg products and 10% of CGF on a flour basis were evaluated against a control (no egg, no CGF). The feasibility of partial baking of rolls was studied on control as well as fresh eggs and carob germ flour formulas during 5 baking times (0, 8, 10, 12 and 18 minutes). Breads were evaluated for crumb and crust color, specific volume, cell profile, Texture Profile Analysis (TPA) and consumer acceptability. Results showed that rolls containing egg ingredients had higher specific volumes than control (p<0.05) with an increase from 1.45 cm[superscript]3/g to 1.96 cm[superscript]3/g. Carob germ flour did not have a significant effect on specific volume. Eggs also improved cell elongation and produced significantly darker crust (p<0.05). CGF did not appear to have an effect on cell elongation but increased average cell number when combined with egg ingredients, and greatly impacted rolls texture. The combination of fresh eggs or egg whites with CGF reduced significantly (p<0.05) crumb hardness from 2,074 to 1,404g and 1,468g of force respectively. Par-baked dinner rolls displayed similar color, volume, cell profile and texture trends to conventionally baked rolls. Sensory study revealed that acceptability, organoleptic characteristics and willingness to buy of par-baked dinner rolls could be similar to that of conventional wheat products. This research proved that the addition of eggs and CGF to a GF rolls formulation resulted in better overall quality of the product. Moreover, par-baking of the rolls showed great potential to provide safe, convenient and acceptable GF foods to celiac individuals.

Das Metabolische Syndrom stellt eine Kombination verschiedener metabolischer Anomalien in einem Individuum dar. Starkes Übergewicht gilt als maßgebende Größe in der Genese des Syndroms, welches mit einem enormen Risiko für kardiovaskuläre Erkrankungen einhergeht. Um die stark steigende Prävalenz des Metabolischen Syndroms einzudämmen, sind dringend Konzepte für die Behandlung, vor allem jedoch für die Prävention von Übergewicht erforderlich. Einen wichtigen Beitrag leisten diesbezüglich Ballaststoffe in der Ernährung. Sie tragen auf unterschiedlichen Wegen zur Gewichtskontrolle bei und beeinflussen zudem verschiedene mit dem Metabolischen Syndrom assoziierte Blutparameter. Ebenso werden protektive Effekte von Polyphenolen, welche zur Gruppe der sekundären Pflanzenstoffe zählen, beschrieben. Diese wirken u. a. auf den Glukose- sowie den Insulinhaushalt und greifen darüber hinaus in die Regulation der Fettverbrennung sowie des Energieverbrauches ein. Die Kombination beider Substanzgruppen verspricht bedeutendes gesundheitsförderndes Potential; dieses wurde gegenwärtig jedoch kaum untersucht. Carobballaststoff ist ein polyphenolreicher und vorwiegend unlöslicher Extrakt der Frucht des Johannisbrotbaumes (Ceratonia siliqua L). Bislang publizierte Studien zur physiologischen Wirksamkeit dieses Ballaststoffpräparates weisen sowohl beim Tier als auch beim Menschen bemerkenswerte hypocholesterinämische Eigenschaften nach. Inwiefern sich der Verzehr des Carobballaststoffes ebenso auf die Entwicklung von Übergewicht sowie anderen Messgrößen des Metabolischen Syndroms auswirkt, ist allerdings nicht bekannt. Die Zielstellung der Promotionsarbeit bestand darin, die postprandialen Wirkungen des Carobballaststoffverzehrs mit Hilfe einer Humanstudie aufzuzeigen. In die randomisierten, einfach verblindeten Untersuchungen im cross-over-Design wurden 20 gesunde Erwachsene im Alter zwischen 22 und 62 Jahren eingeschlossen. Unter Verwendung variierender Begleitmahlzeiten wurden die postprandialen Effekte verschiedener Mengen des Carobballaststoffes untersucht. Hierbei standen die Veränderungen der Plasmakonzentrationen von Glukose, Triglyceriden (TG), totalem und acyliertem Ghrelin sowie der Serumkonzentrationen von Insulin und nicht-veresterten Fettsäuren (NEFA) im Mittelpunkt der Betrachtungen. Der Verzehr des Carobballaststoffes in Kombination mit 200 ml Wasser und 50 g Glukose erhöhte die postprandialen Glukose- und Insulinkonzentrationen gegenüber der Glukoselösung ohne Ballaststoffzusatz. In Kombination mit 400 ml einer Flüssigmahlzeit verzehrt, senkte Carobballaststoff die postprandialen TG-, NEFA- und Ghrelin- (acyliert) Antworten. Die Untersuchung des respiratorischen Quotienten nach Zusatz von Carobballaststoff zur Flüssigmahlzeit mittels indirekter Respirationskalorimetrie bekräftigte die bereits bekannten Effekte auf den Lipidmetabolismus und wies zudem eine Steigerung der Fettverwertung unter Verminderung der Glukoseoxidation nach. Wurde Carobballaststoff schließlich in Lebensmittel eingebracht, sanken nach dem Verzehr dieser Lebensmittel erneut die postprandialen Konzentrationen an TG und NEFA. Gleichzeitig erhöhten sich die Glukose-, Insulin- sowie Ghrelin- (acyliert) Antworten. Carobballaststoff löst in Abhängigkeit von der jeweils verzehrten Begleitmatrix unterschiedliche Effekte aus. Das Präparat weist beachtliche Wirkungen auf die Blutlipide sowie den Energieverbrauch auf, hat indes ungünstige Wirkungen auf die Blutglukose, sofern er in Kombination mit einer veränderten Nährstoffmatrix aufgenommen wird. Carobballaststoff besitzt starkes gesundheitsförderndes Potential; jedoch sind weitere Studien notwendig, um seine Wirkungen sowie deren Voraussetzungen besser zu verstehen. Ferner sollten Untersuchungen über einen längeren Zeitraum vorgenommen werden, um die langfristige Relevanz der gewonnenen Ergebnisse darzulegen. Danach stellt die Anreicherung spezieller Lebensmittel mit Carobballaststoff einen geeigneten Weg dar, um von den viel versprechenden protektiven Wirkungen des Präparates zu profitieren. / The metabolic syndrome is a clustering of metabolic abnormalities and cardiovascular risk factors that occur in one individual. Since many individuals suffering of the metabolic syndrome are overweight, dietary treatment should primarily focus on weight reduction and control to handle and, much more importantly, to prevent the genesis of the syndrome. Dietary fiber could play a role in the management of the metabolic syndrome through its ability to control body weight as well as several parameters associated to the syndrome. Beside this, dietary polyphenols have been shown to influence glucose and insulin metabolism. Furthermore, studies in mice and men showed that polyphenols might increase fat oxidation and energy expenditure. The combination of both, dietary fiber and polyphenols, may exert beneficial health effects, which are not known up to now. A polyphenol-rich insoluble dietary fiber preparation from carob pulp (Ceratonia siliqua L; carob fiber) showed significant hypocholesterolemic activity in different animal trails and two human studies indicating that carob fiber may have potent health effects. However, short term effects on parameters associated with the control of the metabolic syndrome are not known. Therefore, a study was conducted to investigate the postprandial effects of carob fiber in healthy humans using different background food matrices. The study was designed as a randomized single-blind cross-over study in 20 subjects, aged 22 to 62 years. During several examinations plasma glucose, triglycerides (TG), total and acylated ghrelin, cholecystokinin (CCK) and peptide YY (PYY) as well as serum insulin and non-esterified fatty acids (NEFA) were repeatedly assessed before and after ingestion of carob fiber in combination with different test meals. The study results showed that carob fiber, consumed within a glucose load, increased plasma glucose and serum insulin compared to control. Plasma PYY increased after consumption of carob-enriched glucose solution. In a second part of the study, postprandial changes were assessed before and after ingestion of an isocaloric standardized liquid meal with or without carob fiber. Carob fiber intake lowered acylated ghrelin, triglycerides, and NEFA compared to control meal. Postprandial energy expenditure was increased and RQ was reduced after liquid meal with carob fiber compared to control meal. Finally, effects of the consumption of carob fiber-enriched foods were investigated at the subsequent day. Enrichment of foods increased plasma glucose and acylated ghrelin responses. Plasma triglycerides and serum NEFA responses were lower after bread ingestion compared to control. These results indicate that the effects of polyphenol-rich carob fiber on different blood parameters depend on the meal composition. Carob fiber showed beneficial effects on blood lipids as well as energy homeostasis, but also a deteriorated glycemic control when administered within a different food matrix. Further studies are needed to clarify observed effects and the mechanisms behind. Additionally, present study shows only short-term effects of carob fiber consumption, which might be less pronounced after long-term consumption due to adaptation mechanisms. Thus, future studies should elucidate the relevance of observed effects in long-term.

Hydrocolloid stabilizers are commonly used in ice cream formulations to provide body and reduce ice crystal growth during storage. We conducted a retail survey of 65 different vanilla ice cream brands and found the majority of manufacturers primarily use 1 or more of 4 different hydrocolloid sources: guar gum, carrageenan, locust and carob bean gum, or cellulose gum or gel. However, many consumers view hydrocolloids as unnatural, and the presence of hydrocolloids on an ingredient declaration may negatively affect purchase intent. Our survey of 705 consumers showed significant differences in purchase intent for vanilla ice cream, based on ingredient declarations containing different hydrocolloid stabilizers. A response surface central composite design was used to optimize ice cream stability using combinations of tapioca flour, carob bean gum, and citrus fiber, 3 consumer-preferred hydrocolloid stabilizers. Instrumental evaluations considered the dependent variables mix viscosity, ice cream hardness and toughness, melt-rate, and ice crystal size. A trained sensory panel also rated iciness, melt-rate, ease of breakdown in the mouth, and vanilla intensity. Each of the dependent variables from the trained panel and instrumental analysis were measured before and after a 3-week accelerated temperature cycling test. A regression analysis of the central composite design data combined instrumental and trained-panel results to compute a response surface based on the regression equation of each attribute. Using the response surface, 3 different optimized mix formulations were determined. The 3 different mixes were optimized using: 1) all dependent variables evaluated, 2) only sensory iciness scores, and 3) ice crystal size only. An untrained consumer panel evaluated samples before and after temperature cycling test, and rated vanilla ice creams prepared from all 3 optimized mixes against a control ice cream, prepared with a natural commercial stabilizer blend. The uncycled products prepared using optimized stabilizer blend were at statistical parity with the control product for overall acceptance, purchase likelihood, preference, sweetness and vanilla intensity, rate of melting in the mouth, texture and hardness. One or more of the optimized products were rated significantly better than the control for creaminess and texture. For products subjected to temperature cycling, 1 or more of the optimized products were rated significantly better than control for all attributes except sweetness and vanilla flavor intensity. This research indicates that more consumer-preferred options for ice cream stabilization are available to the ice cream industry, with performance and sensory results equal to other commercial hydrocolloid stabilizer blends.

The nutritional value for the following species have been determined: P.pallida, P.laevigata var. andicola, P.flexuosa, P.nigra, P.alba of the genus Proposis. To obtain this purpose surveys were done about the use and consumption of the Carob Tree in the High and Central Valleys of the Cochabamba Department. As a result of the surveys it was determined that the consumption of the pods made up for 54% of human nourishment. The species that were consumed the most were P.pallida and P.laevigata var. andicola. Within the uses of these leguminous pods, the pods are directly consumed in their raw form and on a lesser scale in their cooked form. It was also observed that a minority use it as flour which is then ingested principally in the form of pito and api. Afterwards a chemical bromatologic analysis was done and it was determined that there was a high content of carbohydrates and fiber, a relatively good percentage of protein and a low ethereal extract, standing out more than the minerals calcium and iron. It was also determined that the highest content of protein and ethereal extract show P.laevigata var. andicola and P.nigra, the fiber P.laevigata var. andicola, and P.pallida calories. The species with the most concentration of the minerals phosphorus, iron, and calcium were P.pallida and P.nigra. The biological tests for the protein quality performed for the two species that were the most consumed show poor results of Real Digestibility (Dr), Net Use of Real Protein (UNPr), Real Biological Value (VBr), and the Relation of the Efficiency of Protein (PER). These are considerably becoming better through a thermic process (toasted 10min). The leguminous cereal complementation (corn-carob tree), favorably increased all of the biological indexes. The species P.pallida showed superiority compared to the results obtained for P.leavigata var. andicola. In the meantime these species can be considered a good energy source and with the combination of cereals can effectively cover caloric protein deficiencies.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The whey has been gaining ground and importance in the food industry. The milk beverages are one way to use this coproduct, the ones with chocolate flavor has broad consumer acceptance of both sexes and various ages. Carob is being studied as a possible replacement for cocoa by presenting color, odour and taste similar to cocoa, have lower production costs and does not contain caffeine and theobromine, substances known as antinutritional factors of cocoa. Given these facts, it was aimed to the development of milk beverages with the use of whey powder and partial replacement of cocoa for carob. The results showed that the use of 15% of whey powder added nutritional value and obtained the highest preference in the sensorial analysis, the use carob as a substitute for cocoa do not interfere in the physicalchemical analyzes of milk beverages. Were prepared 12 formulations of milk beverages with different concentrations of cocoa and carob powder, and analysis microbiological, sensory and physicochemical were performed. The counts of micro-organisms remained within the required by Brazilian law, the acceptability of sensory analysis revealed that higher levels of cocoa and carob added to milk beverages, increase the acceptance of evaluative attributes (color, odour, flavor and viscosity) the milk drink with higher level of carob powder was preferred over the other milk beverages. There were no statistical differences (p> 0.05) on the parameters of fat, moisture, protein and reducing sugars, the milk beverages had shelf life of 14 days. Considering these results it can be concluded that the partial replacement of cocoa for carob in milk beverages is an attractive alternative, since the samples were well accepted sensorially, had good intentions to purchase and there were no losses in nutritional formulations of milk beverages. / O lactossoro vem ganhando espaço e importância na indústria alimentícia. As bebidas lácteas são uma das formas de aproveitamento desse co-produto, estas, com sabor chocolate têm ampla aceitação pelos consumidores, de ambos os sexos e de várias faixas etárias. A alfarroba está sendo estudada como um possível substituto do cacau, por apresentar cor, odor e sabor semelhantes ao cacau, apresentar menor custo de produção e não conter cafeína e teobromina, substâncias ditas como fatores antinutricionais do cacau. Tendo em vista esses fatos, objetivou-se a elaboração de bebidas lácteas visando o uso de lactossoro em pó e a substituição parcial do cacau por alfarroba. Os resultados mostraram que a utilização de 15% de soro em pó agregou valor nutricional e obteve a maior preferência na análise sensorial; a utilização de alfarroba como substituta do cacau não interferiu nas análises físico-químicas das bebidas lácteas. Elaborou-se 12 formulações de bebidas lácteas com diferentes concentrações de cacau e alfarroba em pó, análises microbiológicas, sensoriais e físicoquímicas foram realizadas. As contagens de micro-organismos mantiveram-se dentro do exigido pela legislação brasileira; a análise sensorial de aceitabilidade revelou que quanto maiores os teores de cacau e alfarroba adicionados às bebidas lácteas, aumentava-se a aceitação dos atributos avaliados (cor, odor, sabor e viscosidade) a bebida láctea com maior teor de alfarroba em pó foi a preferida em relação às demais bebidas lácteas. Não foram observadas diferenças estatísticas (p > 0,05) nos parâmetros de gordura, umidade, proteína e açúcares redutores, as bebidas lácteas apresentaram vida de prateleira de 14 dias. Considerando os resultados obtidos pode-se concluir que a substituição parcial do cacau por alfarroba em bebidas lácteas é uma alternativa atrativa, visto que as amostras foram bem aceitas sensorialmente, obtiveram boa intenção de compra e não ocorreram perdas nutricionais nas formulações de bebidas lácteas.

Cette thèse de doctorat porte sur la valorisation du déchet solide issu de la préparation de la mélasse de caroube libanaise pour la production de bioénergie et de molécules à valeur ajoutée. L’analyse de la composition de ce déchet a montré qu’il contient 45 % (g/g) de sucres, substrats exploitables pour la fermentation alcoolique ou lactique, la production de biohydrogène, ou comme source de carbone pour la croissance d’une algue dans un procédé de phycoremédiation (traitement des eaux par les algues) pour l’élimination de micropolluants pharmaceutiques. Les résultats obtenus ont montré que la fermentation alcoolique en phase liquide à partir d’extraits de déchet de caroube peut conduire à des rendements élevés en bioéthanol sous réserve d’enrichir le milieu de culture par les éléments nécessaires pour la croissance de la levure Saccharomyces cerevisiae (en particulier l’azote), tandis qu’il est possible de produire directement le bioéthanol sur le déchet par fermentation en milieu solide sous réserve de contrôler précisément l’humidité. Nous avons également démontré que la fermentation lactique par immobilisation de Lactobacillus rhamnosus sur des billes d’alginate constitue une alternative possible au bioéthanol pour les sucres extraits du déchet. Cependant, un enrichissement du milieu de culture, ainsi que l’utilisation d’une invertase en prétraitement sont nécessaires afin de maximiser le rendement et la productivité de l’acide lactique. L’immobilisation des microorganismes a permis de réutiliser les mêmes billes d’alginate au cours de cinq cycles successifs de production. Une autre alternative potentielle aux voies précédentes est la fermentation sombre pour la production de biohydrogène directement à partir du déchet. Si des rendements intéressants ont été atteints, il faut noter que comme précédemment dans le cas de la fermentation solide du déchet, une forte dépendance aux conditions initiales de broyage a été mise en évidence ; de plus, à la carence en azote qui obligeait à supplémenter les milieux en fermentation en phase liquide s’ajoutent des besoins en fer. Enfin, l’utilisation du déchet comme substrat carboné dans un procédé de phycoremédiation avec l’algue Ankistrodesmus braunii a montré que l’élimination de 90% du diclofénac initial pouvait être atteinte en conditions de mixotrophie, même si seulement un tiers du diclofénac éliminé est effectivement métabolisé par l’algue. / This PhD thesis deals with the valorization of the solid waste generated by the Lebanese carob molasse process with the aim to produce bioenergy or high added-value platform molecules. The chemical analysis of this waste has highlighted that it contains 45% (w/w) sugars, which can be used as a substrate for alcoholic or lactic fermentation, the production of biohydrogen by dark fermentation, or as a carbon source for the growth of microalgae in a phycoremediation process (water treatment process based on algae/microalgae) for the removal of pharmaceutical micropollutants. Experimental results have shown that high ethanol yield and productivity could be achieved through alcoholic fermentation in the liquid phase using extracts from carob waste as the substrate, provided the culture medium was enriched by complementary nutrients (especially a nitro-gen source) to enhance the growth of the yeast Saccharomyces cerevisiae; in parallel, the direct production of ethanol from the solid waste based on solid-state fermentation was also proved to be efficient, provided humidity was accurately controlled. In addition, lactic fermentation per-formed with immobilized Lactobacillus rhamnosus on alginate beads was shown to constitute a potential alternative to bioethanol for the extracts from carob waste. An enriched culture medium was, however, necessary, together with the use of an invertase enzyme as a pretreatment so as to maximize the yield and the productivity of lactic acid. Using microorganism immobilization lead to the opportunity to recycle at least five times alginate beads in successive culture cycles. Another potential alternative to the above-mentioned valorization pathways was proved to lie in the dark fermentation process for biohydrogen production, directly using the solid waste. While attractive yields were reached, a strong dependence on the particle size from grinding pretreatment was emphasized, as in solid-state fermentation; moreover, iron supplementation in the culture medium was compulsory, in addition to nitrogen supplementation already described when fermentation was carried out in the liquid phase. Finally, when the carob waste was used as a substrate for the growth of the algae Ankistrodesmus braunii in a phycoremediation process, a removal yield of the initial diclofenac content about 90% could be reached under mixotrophic conditions, even though only one third of the diclofenac removed was effectively metabolized.