Bioconversion of Cheese Whey and Food By-Products by Phaeodactylum tricornutum into Fucoxanthin and n-3 Lc-PUFA through a Biorefinery Approach.

This study investigates the potential of utilizing three food wastes: cheese whey (CW), beet molasses (BM), and corn steep liquor (CSL) as alternative nutrient sources for the cultivation of the diatom Phaeodactylum tricornutum, a promising source of polyunsaturated eicosapentaenoic acid (EPA) and the carotenoid fucoxanthin. The CW media tested did not significantly impact the growth rate of P. tricornutum; however, CW hydrolysate significantly enhances cell growth. BM in cultivation medium enhances biomass production and fucoxanthin yield. The optimization of the new food waste medium was conducted through the application of a response surface methodology (RSM) using hydrolyzed CW, BM, and CSL as factors. The results showed a significant positive impact of these factors (p < 0.005), with an optimized biomass yield of 2.35 g L-1 and a fucoxanthin yield of 3.64 mg L-1 using a medium composed of 33 mL L-1 of CW, 2.3 g L-1 of BM, and 2.24 g L-1 of CSL. The experimental results reported in this study showed that some food by-products from a biorefinery perspective could be utilized for the efficient production of fucoxanthin and other high-added-value products such as eicosapentaenoic acid (EPA).

Effect of Salt Addition and Fermentation Time on Phenolics, Microbial Dynamics, Volatile Organic Compounds, and Sensory Properties of the PDO Table Olives of Gaeta (Italy).

'Oliva di Gaeta' is almost certainly the most important and well-known PDO denomination for table olives in Italy. Their production is based on a specific two-stage trade preparation called the 'Itrana' method. In this work, we investigated how variations in the duration of the initial water fermentation (i.e., 15 and 30 days) and the salt concentration (i.e., 6% and 8% NaCl) influence the chemical features, microbial dynamics, polyphenols, volatile organic compounds, and sensory features of 'Oliva di Gaeta'. The time of the addition of salt did not affect the final concentration in the brine, but a longer initial water fermentation (before salt addition) led to lower pH values. The bacterial count constantly increased until the salt addition (i.e., either 15 or 30 days), while the yeast population peaked on day 30. Generally, the two different salt concentrations did not affect the count of microorganisms at the end of fermentation, with the only exception being a higher lactic acid bacteria count for the treatment with 6% salt added at 30 days. At commercial maturity, the crucial bitter tastant oleuropein was not completely removed from the drupes, and differences in salt concentration and the length of the first-stage water fermentation did not influence its content at the end of olive curing. Richer volatile profiles of olives were detected with higher-salt treatments, while the combination of low salt and early saline treatment provided a more distinct profile. Longer initial water fermentation caused a small increase in some phenolic compounds (e.g., iso-verbascoside, verbascoside, and hydroxytyrosol-glucoside). A panel test indicated that salt application at 30 days resulted in a more "Sour" and "Bitter" taste, irrespective of the salt concentration. The low salt concentration coupled with the late saline treatment resulted in more "Fruity" notes, probably due to the higher production of esters by lactobacilli. The slightly bitter perception of the olives was consistent with the partial removal of oleuropein. Our work revealed the characteristics of the 'Itrana' method and that the variation in salt concentration and its time of application changes parameters ranging from the microbial dynamics to the sensory profile. Specifically, our data indicate that 6% NaCl coupled with a longer initial water fermentation is the most different condition: it is less effective in blocking microbial growth but, at the same time, is more potent in altering the nutritional (e.g., polyphenols) and sensorial qualities (e.g., bitterness and fruitiness) of 'Oliva di Gaeta'.

Formulation of New Media from Dairy and Brewery Wastes for a Sustainable Production of DHA-Rich Oil by Aurantiochytrium mangrovei.

Mozzarella stretching water (MSW) is a dairy effluent generated from mozzarella cheese production that does not have a real use and is destined to disposal, causing environmental problems and representing a high disposal cost for dairy producers. Spent brewery yeast (SBY) is another promising food waste produced after brewery manufacturing that could be recycled in new biotechnological processes. Aurantiochytrium mangrovei is an aquatic protist known as producer of bioactive lipids such as omega 3 long chain polyunsaturated fatty acids (ω3 LC-PUFA), in particular docosahexaenoic acid (DHA). In this work MSW and SBY have been used to formulate new sustainable growth media for A. mangrovei cultivation and production of DHA in an attempt to valorize these effluents. MSW required an enzymatic hydrolysis to enhance the biomass production. The new media obtained from hydrolysed MSW was also optimized using response surface methodologies, obtaining 10.14 g L-1 of biomass in optimized medium, with a DHA content of 1.21 g L-1.

Biophenolic Compounds Influence the In-Mouth Perceived Intensity of Virgin Olive Oil Flavours and Off-Flavours.

In this study, the influence of phenolic compounds on the sensory scores attributed to extra virgin olive oil (EVOO) by panel test was investigated. Two model olive oils (MOOs) with identical concentrations of volatile compounds, differing only in the amount of biophenols (297 vs. 511 mg kg-1), were analysed by two official panels and by SPME-GC/MS. Six other MOOs set up by the two previous models were also tested and analysed. They were formulated separately with the addition of three off-flavours ('rancid', 'winey-vinegary' and 'fusty-muddy'). While high levels of EVOO phenolic compounds did not produce any effect on the headspace concentration of volatile compounds, they did affect the scores of both positive and negative sensory attributes of EVOO, due to the well-known in-mouth interactions between EVOO phenols, saliva and volatile compounds. In particular, a decrease of about 39% in the positive fruity score was found in the presence of a higher concentration of phenols. Regarding EVOO off-flavours, the higher level of phenolic compounds decreased by about 23% the score of 'fusty-muddy' defect and increased the score of 'winey-vinegary' defect about 733%. No important effect of EVOO phenolics on the perceived intensity of the 'rancid' defect was found. These findings could be helpful in explaining some discrepancies of panel test responses observed during extra virgin olive oil shelf life.

Effects of Inclusion of Fresh Forage in the Diet for Lactating Buffaloes on Volatile Organic Compounds of Milk and Mozzarella Cheese.

This paper aimed to assess the effects of feeding fresh forage on volatile organic compounds (VOCs) of buffalo milk and mozzarella cheese. Sixteen lactating buffaloes were equally allotted into two groups fed diets containing (experimental (Exp) group) or not (control (Ctl) group) 20 kg/d of fresh sorghum. Milk from the groups was separately collected and transformed in the traditional 'Mozzarella di Bufala Campana' Protected Denomination of Origin (PDO). Three batches of mozzarella were produced for each diet and they were analyzed, along with the two bulks of milk, for VOC composition, by using solid phase micro-extraction (SPME) coupled with gas-chromatography/mass spectrometry (GC/MS). The use of fresh forage increased the levels of long chain fatty acids along with the contents of aldehydes, and this could be responsible for an increase in green notes of milk. The use of the Ctl diet, containing a higher proportion of silage, increased the ketones, acids, and esters, which are compounds that could raise the cheese and fruity notes of milk. The mozzarella was less affected by the dietary treatment than milk. The use of fresh forage (sorghum) enhanced the green notes of milk and induced a few changes in the VOC profile of the typical PDO Mozzarella di Bufala Campana cheese, that were nonetheless detectable by sensory analysis. The low level found for butanoic acid, 2,3-pentanedione, and propyl acetate in mozzarella cheese obtained with fresh forage diet can lead to perceive less the olfactory notes of cheese, cream, and fruit.

Volatile compounds, physicochemical and sensory characteristics of Colatura di Alici, a traditional Italian fish sauce.

BACKGROUND: The present study reports for the first time the physical, chemical and sensory characterization of a traditional fish sauce from Italy, called 'Colatura di Alici', which is considered to be the direct descendant of the ancient roman sauce 'Garum'. RESULTS: Among the volatile compounds, carboxylic acids, aldehydes, sulphuric compounds and pyrazines were identified. The most abundant compounds identified were 3-methyl-butanoic acid, nonanal and 3-methyl-butanal. The sample A had the highest amount of volatile fatty acids (>250 µg kg-1 ) and sample B had the highest concentration of aldehydes (>80 µg kg-1 ). These volatile compounds arise from the long fermentation activity and oxidation of fatty acids of the fish flesh and are responsible for the strong cheesy and fishy aroma. The protein content ranged from 90 to 130 g kg-1 , and the soluble solids ranged from 30 to 38 °Brix, as a result of the high amount of NaCl used during processing. The sensory analysis showed a great intensity for the cheesy, fishy and umami attributes, and a medium intensity for the roasted and meaty attributes. CONCLUSION: For the first time, the volatile compounds and sensory characteristics of this traditional fish sauce are reported. The samples differ in terms of sensory and aromatic profiles. These differences are probably linked to the poorly standardized methods used in the production process. Overall, the results of the present study could be used by local fish sauce producers to monitor the quality of the product and to improve the production process. © 2020 Society of Chemical Industry.

Volatile Organic Compound and Fatty Acid Profile of Milk from Cows and Buffaloes Fed Mycorrhizal or Nonmycorrhizal Ensiled Forage.

Seed inoculation of forage crops by arbuscular mycorrhizal fungi (AMF) generally results in higher profitability, but also modifies the chemical composition of silage in terms of increased biomass, protein, and dry matter. Raw milk aroma is affected by the type of feed. This work investigated the influence of ensiled forage obtained by seed inoculation with AMF on the volatile fractions and fatty acid composition of milk. Two experiments were carried out: in the first, buffaloes were fed maize silage, and in the second, cows were fed sorghum silage. The volatile fractions of milk were quantified by headspace solid-phase micro-extraction (HS-SPME), combined with gas chromatography-mass spectrometry (GC-MS), and fatty acids by gas chromatography (GC). The ensiled forage obtained with AMF increased saturated fatty acids (SFAs), and decreased monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) in both experiments. The volatile fraction in milk samples obtained from bovines fed mycorrhizal ensiled forage showed an increase of free fatty acids and ketones, responsible for cheesy and fruity odors. Aldehydes, responsible for green and grassy notes, increased only in the milk from buffaloes fed ensiled maize. Our results suggest that inoculation of maize and sorghum seed with AMF, combined with a low rate of fertilizers, leads to ensiled forage that could slightly affect the FA profile and odor quality of milk.

Use of odorant series for extra virgin olive oil aroma characterisation.

BACKGROUND: Extra virgin olive oil (EVOO) volatile composition is mainly used as a means of characterisation and authentication, especially for protected denomination of origin (PDO) products. This work investigated the volatile compounds from 25 EVOOs from four Spanish (Cornicabra, Manzanilla Castellana, Picual and Manzanilla Cacereña) and four Italian (Ortice, Ravece, Nocellara del Belice and Itrana) cultivars in terms of odour activity value (OAV). Forty-seven volatile compounds were analysed by solid phase microextraction gas chromatography/mass spectrometry (SPME-GC/MS). OAVs of volatile compounds with similar descriptors were grouped in order to establish eight odorant series: fruity, grass, apple, tomato, floral, woody-spicy, fatty and mushroom. RESULTS: No differences in sensory descriptors were observed among the EVOOs analysed by official VOO sensory analysis. The method of odorant series applied herein was demonstrated to successfully characterise EVOO odour as expected from a sensory panel but using only instrumental analysis of volatile compounds, and giving additional reliable quantitative information. The results can be presented as a 'barcode', providing a visual and effective graphical representation allowing an easy and rapid description of EVOO sensory attributes using instrumental data. CONCLUSION: The odorant series have the potential to better differentiate the aroma of food products, opening new possibilities allowing a schematic and effective visual representation to be used for EVOO quality control and consumer information, especially in new olive oil consuming countries. © 2018 Society of Chemical Industry.

Extra virgin olive oil aroma release after interaction with human saliva from individuals with different body mass index.

BACKGROUND: The interindividual variability observed in saliva characteristics raises the question of its relationship with variability in fat sensory perception, particularly in aroma compounds. In the present study, which aimed to measure aroma release from different individuals, eleven key aroma compounds of extra virgin olive oil (EVOO) were monitored and quantified in dynamic headspace after an in vitro interaction between EVOO and human saliva. Therefore, 60 individuals were studied from those who were normal weight (NW), overweight (OW) and obese (O). RESULTS: OW and O demonstrate a higher release of C6 compounds compared to NW. By contrast, NW have a higher release of C5 compounds. Pentanal and hexanal also increased after saliva interaction in a refined olive oil that is free from volatiles. Among the saliva samples with a higher release in NW individuals, only pentanal was different. However, the low levels of these lipid oxidation end-products do not appear to be very important with respect to increasing odorous fat sensitivity. CONCLUSION: The results obtained in the present study demonstrate the important role of saliva in the aroma release of EVOO, indicating that humans can perceive it differently in relation to their body mass index. © 2017 Society of Chemical Industry.

Fruit position within the canopy affects kernel lipid composition of hazelnuts.

BACKGROUND: The aim of this research was to study the variability in kernel composition within the canopy of hazelnut trees. RESULTS: Kernel fresh and dry weight increased linearly with fruit height above the ground. Fat content decreased, while protein and ash content increased, from the bottom to the top layers of the canopy. The level of unsaturation of fatty acids decreased from the bottom to the top of the canopy. Thus, the kernels located in the bottom layers of the canopy appear to be more interesting from a nutritional point of view, but their lipids may be more exposed to oxidation. The content of different phytosterols increased progressively from bottom to top canopy layers. CONCLUSION: Most of these effects correlated with the pattern in light distribution inside the canopy. The results of this study indicate that fruit position within the canopy is an important factor in determining hazelnut kernel growth and composition. © 2017 Society of Chemical Industry.

Healthy virgin olive oil: a matter of bitterness.

Virgin olive oil (VOO) is the pillar fat of Mediterranean diet. It is made from olive fruits and obtained by squeezing olives without any solvent extraction. Respect to the seed oils, an unique polar polyphenol-rich fraction gives VOO a bitter and pungent taste. The recent substantiation by European Food Safety Authority (EFSA) of a health claim for VOO polyphenols may represent an efficient stimulus to get the maximum health benefit from one of the most valuable traditional product of Mediterranean countries educating consumers to the relationship between the VOO bitterness and its health effect. Agronomical practices and new processing technology to avoid phenolic oxidation and hydrolysis and to enhance the aromatic components of the VOO have been developed and they can be used to modulate taste and flavor to diversify the products on the market. VOOs having high concentration of phenol compounds are bitter and pungent therefore many people do not consume them, thus loosing the health benefits related to their intake. In this paper, the chemist's and nutritionist's point of view has been considered to address possible strategies to overcome the existing gap between the quality perceived by consumer and that established by expert tasters. Educational campaigns emphasizing the bitter-health link for olive oils should be developed.

Extra virgin olive oil: from composition to "molecular gastronomy".

The aim of this chapter is to provide a brief overview of the recent results of studies on extra virgin olive oil (EVOO) and its interactions with other food ingredients during cooking, to highlight basic molecular aspects of the "magic" of EVOO and its role in Mediterranean gastronomy. The use of raw EVOO added to foods after cooking (or as a salad oil) is the best way to express the original flavour and to maximize the intake of natural antioxidants and compounds related to positive effects on human health (hypotensive, anti-inflammatory, and anti-cancerogenic, among others). EVOO, however, also exhibits its protective properties during/after cooking. Different chemical interactions between biophenolic compounds and other food ingredients (water, milk proteins, carotenoids of tomato, omega-3 polyunsaturated fatty acids in canned-in-oil fish and meat or fish proteins) occur. Even during cooking, EVOO exhibits strong antioxidant properties and influences the overall flavour of cooked foods. The physical (partitioning, emulsion) and chemical (hydrolysis, covalent binding, antioxidant properties) phenomena occurring during cooking of EVOO are discussed with emphasis on the changes in the sensory (bitterness and fruity flavour) and nutritional qualities of some traditional Mediterranean foods. In particular, tomato-oil interactions during cooking, fish canning in EVOO, meat marinated in EVOO before cooking and roasting and frying in EVOO are examined. The interactions between EVOO antioxidants and flavours with milk proteins are also briefly discussed.

From flower to honey bouquet: possible markers for the botanical origin of Robinia honey.

Flowers are complex structures devoted to pollinator attraction, through visual as well as chemical signals. As bees collect nectar on flowers to produce honey, some aspects of floral chemistry are transferred to honey, making chemical markers an important technique to identify the botanical and geographical origins of honey. We applied a new approach that considers the simultaneous analysis of different floral parts (petals, stamens + pistils, calyxes + nectarines, and nectar) and the corresponding unifloral honey. We collected fresh flowers of Robinia pseudoacacia L. (black locust), selected five samples of Robinia honey from different geographical origins, applied SPME-GC/MS for volatile analyses, and defined the chemical contribution added by different floral parts to the honey final bouquet. Our results show that honey blends products from nectar as well as other flower parts. Comparing honey and flower profiles, we detected compounds coming directly from flower parts but not present in the nectar, such as hotrienol and ß-pinene. These may turn out to be of special interest when selecting floral markers for the botanical origin of honey.

Influence of some environmental factors on drupe maturation and olive oil composition.

BACKGROUND: Understanding the relationships between olive cultivars and the cultivation environment as well as optimising cultivation choices can lead to maximum expression of oil production in terms of both quantity and quality. For this purpose, samples of the Ortice olive cultivar grown in two different environments in southern Italy at altitudes of 500 and 50 m above sea level (a.s.l.) were harvested on various dates to monitor drupe maturation and determine the nutritional and chemical characteristics of the oils. RESULTS: Fruits grown at 50 m a.s.l. ripened about 10-15 days earlier than those grown at 500 m a.s.l. The oil obtained at 500 m a.s.l. was characterised by a higher content of total polyphenols and a higher content of oleic and stearic acids. CONCLUSION: The different heat accumulation related to the environment affected drupe development as well as the colouring trend and oil content. Consequently, the growth environment changed the content of Ortice oil fatty acids and polyphenols, while the flavour profile remained fairly stable in both environments, with the cultivar effect prevailing over the environment factor.

Characterization and evaluation of olive germplasm in southern Italy.

BACKGROUND: The southern Italian region of Campania has suitable pedo-climatic conditions and a large varietal heritage able to produce oils with high typicality. The aim of this study was to characterize 20 cultivars belonging to Campania's olive germplasm, evaluating their vegetative and production aspects and their oil quality characteristics. The study was conducted from 2003 to 2009, observing the following aspects in six plants per variety: entry into production, vigour, ripening and drupe oil content. The following analyses were carried out on monovarietal oils, obtained by microextractor: acid composition, polyphenol content and aromatic profile. RESULTS: The agronomic results showed early entry into production for the cultivars Racioppella, Ortolana, Biancolilla and Carpellese. However, entry into production was delayed for Ritonnella, Ortice, Cornia and Rotondella. As regards vegetative behaviour, Asprinia, from the province of Caserta and Pisciottana and Carpellese, from the province of Salerno, proved the most vigorous cultivars. CONCLUSION: Groups of similar cultivars emerged from chemical and sensory analysis of their oils, while other accessions were well characterized and separated from each other, showing a high level of diversity and specificity.

Functional ice cream health benefits and sensory implications.

The increased interest in foods which improve human health and nutrition has led to the need for academia and food industry to invest in the development of innovative and functional ice cream. Since ice cream is the most popular frozen dairy product, it shows good potential to help people improve their diets by reducing the intake of certain nutrients associated with the increased risk of obesity and other related diseases. Moreover, it supplies beneficial and essential components. However, the reduction or removal of common ice cream ingredients or the addition of uncommon ingredients to the standard formulation must not spoil the sensory characteristics and storage stability of ice cream. Hence, to keep the appeal of ice cream high, the food industry is subjected to constant challenges, since functional ice cream must be tasty as well as beneficial. The review focuses on the healthy aspects and sensory implications of ice cream functionalization. In the first part, different replacing strategies such as the substitution of fat and sugar are discussed. In the second part, ice cream enriched with functional ingredients is debated.

Particle size and variety of coffee used as variables in mitigation of furan and 2-methylfuran content in espresso coffee.

In this study, the concentration of furan and 2-methylfuran in espresso coffee (EC) obtained from Arabica and Robusta coffee varieties was determined as a function of specific particle size. The particle size and coffee variety significantly influenced the level of furan and 2-methylfuran. In Arabica variety, furan and 2-methylfuran level increased with increasing particle size. Particularly, from C<200µm to C>425µm fractions, furan increased from 68.27 to 91.48 ng mL-1 while 2-methylfuran from 404.31 to 634.64 ng mL-1. In Robusta variety, the highest concentration of furan and 2-methylfuran occurred in ECs prepared using C300-425µm fraction showing values of 116.39 ng mL-1 and 845.14 ng mL-1, respectively, for furan and 2-methylfuran. On the basis of this experiment, it is possible to establish a mitigation strategy by manipulating the particle size and coffee variety in order to reduce the level of furan and 2-methylfuran in EC up to 11.4% and 18.8%, respectively.

Sustainable production of food grade omega-3 oil using aquatic protists: Reliability and future horizons.

Biotechnological production of omega-3 polyunsaturated fatty acids (PUFAs) has become a commercial alternative to fish oil in the past twenty years. Compared to PUFA production by fatty fishes, that from microorganisms has increased due to its promising sustainability and high product safety and to increasing awareness in the expanding vegan market. Although autotrophic production by microalgae seems to be more sustainable in the long term, to date most of the microbial production of omega-3 is carried out under heterotrophic conditions using conventional fermentation technologies. The present review critically analyzes the main reasons for this discrepancy and reports on the recent advances and the most promising approaches for its future development in the context of sustainability and circular economy.

An Endophytic Fungi-Based Biostimulant Modulates Volatile and Non-Volatile Secondary Metabolites and Yield of Greenhouse Basil (Ocimum basilicum L.) through Variable Mechanisms Dependent on Salinity Stress Level.

Salinity in water and soil is one of the major environmental factors limiting the productivity of agronomic and horticultural crops. In basil (Ocimum basilicum L., Lamiaceae) and other Ocimum species, information on the plant response to mild salinity levels, often induced by the irrigation or fertigation systems, is scarce. In the present work, we tested the effectiveness of a microbial-based biostimulant containing two strains of arbuscular mycorrhiza fungi (AMF) and Trichoderma koningii in sustaining greenhouse basil yield traits, subjected to two mild salinity stresses (25 mM [low] and 50 mM [high] modulated by augmenting the fertigation osmotic potential with NaCl) compared to a non-stressed control. The impact of salinity stress was further appraised in terms of plant physiology, morphological ontogenesis and composition in polyphenols and volatile organic compounds (VOC). As expected, increasing the salinity of the solution strongly depressed the plant yield, nutrient uptake and concentration, reduced photosynthetic activity and leaf water potential, increased the Na and Cl and induced the accumulation of polyphenols. In addition, it decreased the concentration of Eucalyptol and ß-Linalool, two of its main essential oil constituents. Irrespective of the salinity stress level, the multispecies inoculum strongly benefited plant growth, leaf number and area, and the accumulation of Ca, Mg, B, p-coumaric and chicoric acids, while it reduced nitrate and Cl concentrations in the shoots and affected the concentration of some minor VOC constituents. The benefits derived from the inoculum in term of yield and quality harnessed different mechanisms depending on the degree of stress. under low-stress conditions, the inoculum directly stimulated the photosynthetic activity after an increase of the Fe and Mn availability for the plants and induced the accumulation of caffeic and rosmarinic acids. under high stress conditions, the inoculum mostly acted directly on the sequestration of Na and the increase of P availability for the plant, moreover it stimulated the accumulation of polyphenols, especially of ferulic and chicoric acids and quercetin-rutinoside in the shoots. Notably, the inoculum did not affect the VOC composition, thus suggesting that its activity did not interact with the essential oil biosynthesis. These results clearly indicate that beneficial inocula constitute a valuable tool for sustaining yield and improving or sustaining quality under suboptimal water quality conditions imposing low salinity stress on horticultural crops.

Real-life anti-vascular endothelial growth factor treatment for age-related macular degeneration and diabetic macular edema in an Italian tertiary referral hospital.

PURPOSE: To study the real-life outcomes of intravitreal anti-vascular endothelial growth factor treatment in patients affected by age-related macular degeneration and diabetic macular edema in an Italian tertiary referral hospital over a 2-year follow-up. METHODS: Patients with confirmed diagnosis of age-related macular degeneration or diabetic macular edema and 2 years of follow-up were retrospectively analyzed. They underwent a loading dose of three monthly anti-vascular endothelial growth factor injections, with re-treatment performed following a "pro-re-nata" regimen. Best-corrected visual acuity and central retinal thickness were collected and statistically analyzed. RESULTS: In total, 167 diabetic macular edema eyes and 108 age-related macular degeneration eyes were included. Mean age was 63.4 ± 11.8 years for diabetic macular edema and 75.6 ± 8.4 years for age-related macular degeneration. For diabetic macular edema patients, mean number of injections was 5.0 ± 1.7 at 1 year and 2.8 ± 1.8 at 2 years. Mean best-corrected visual acuity improved from 60 Early Treatment for Diabetic Retinopathy Study letters to 66.7 letters at 1 year, and to 70 letters at 2 years (p < 0.001). Mean central retinal thickness decreased from 459 ± 148 µm to 327 ± 163 µm and 261 ± 89 µm, respectively, at the first and the second year (p < 0.001).With respect to age-related macular degeneration patients, mean number of injections was 5.3 ± 2.2 at 1 year and 3.3 ± 1.5 at 2 years. Mean best-corrected visual acuity improved from 63 Early Treatment for Diabetic Retinopathy Study letters to 68 letters at 1 year and to 70 letters at 2 years (p < 0.001). Mean central retinal thickness was 411 ± 146 µm, decreasing to 271 ± 93 µm at 1 year and 260 ± 68 µm at 2 years (p < 0.001). CONCLUSIONS: Our study described the 2-year real-life outcome of anti-vascular endothelial growth factor treatment of diabetic macular edema and age-related macular degeneration.