Effects of Defoliation on Phenolic Concentrations, Antioxidant and Antibacterial Activity of Grape Skin Extracts of the Varieties Blaufränkisch and Merlot (Vitis vinifera L.)

Bibliographic Details
Title:	Effects of Defoliation on Phenolic Concentrations, Antioxidant and Antibacterial Activity of Grape Skin Extracts of the Varieties Blaufränkisch and Merlot (Vitis vinifera L.)
Authors:	Valentina Pavić, Toni Kujundžić, Marina Kopić, Vladimir Jukić, Ulrike Braun, Florian Schwander, Mato Drenjančević
Source:	Molecules, Vol 24, Iss 13, p 2444 (2019)
Publisher Information:	MDPI AG, 2019.
Publication Year:	2019
Collection:	LCC:Organic chemistry
Subject Terms:	defoliation, Blaufränkisch, Merlot, phenols, antioxidant activity, antibacterial activity, Organic chemistry, QD241-441
More Details:	Vitis vinifera L. has been highlighted by its many positive effects on human health, including antibacterial, anti-inflammatory and antioxidant activity due to its rich phytochemical content. Removing six basal leaves has great potential to influence the production of phytochemicals. The purpose of this study was to determine the impact of different terms of defoliation on the antioxidant and antibacterial activity of grape skin extracts of the Blaufränkisch and Merlot varieties. The total phenolic concentration, total and individual anthocyanin concentration, antioxidant activity and antibacterial activity on gram-positive and gram-negative human pathogens have been determined. The study was conducted on three treatments (control treatment without defoliation, defoliation immediately after bloom and defoliation before the start of the véraison phase). The results showed statistically significant enhancement of the total phenolic concentration as well as the antioxidant and antibacterial activity in both studied cultivars. Defoliation just after blooming was the preferable defoliation term in the Merlot variety for achieving the highest total anthocyanin concentration, antioxidant activity and significant increase of antibacterial activity against all four investigated bacteria. Defoliation before the start of the véraison phase was the preferable defoliation term for achieving the highest total anthocyanin concentration in the Blaufränkisch variety. In general, treatment of defoliation immediately after bloom was more beneficial compared with the defoliation before the start of the véraison phase.
Document Type:	article
File Description:	electronic resource
Language:	English
ISSN:	1420-3049
Relation:	https://www.mdpi.com/1420-3049/24/13/2444; https://doaj.org/toc/1420-3049
DOI:	10.3390/molecules24132444
Access URL:	https://doaj.org/article/f38842e967dd4472b0fb67151263d5bd
Accession Number:	edsdoj.f38842e967dd4472b0fb67151263d5bd
Database:	Directory of Open Access Journals
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FullText	Links: – Type: pdflink Url: https://content.ebscohost.com/cds/retrieve?content=AQICAHjPtM4BHU3ZchRwgzYmadcigk49r9CVlbU7V5F6lgH7WwHDyCQB7AYxxW53BRpdTv1mAAAA4jCB3wYJKoZIhvcNAQcGoIHRMIHOAgEAMIHIBgkqhkiG9w0BBwEwHgYJYIZIAWUDBAEuMBEEDMdRZFuw3PU-P962RwIBEICBmmV5FfWJUf60w_I5Sg6lfY1DmkyehGG2WY4L4ThflQaWXXluJnSj_SVu2GT9YHzw0qT_C4VdirlvfnXaAwjt5u5Jqft3PUKjrytaFoR1jgfuUSsN1dboNzEHNdm7z7FxPQfcs4fYXgaGU63HfI9FyaXkpBn8_bN0aJjQuV71RqaogXTwiM-4O_axbjMuUrfpp5hJ_WfSWikIjvU= Text: Availability: 1 Value: <anid>AN0137474727;nnv01jul.19;2019Jul16.05:03;v2.2.500</anid> <title id="AN0137474727-1">Effects of Defoliation on Phenolic Concentrations, Antioxidant and Antibacterial Activity of Grape Skin Extracts of the Varieties Blaufränkisch and Merlot (Vitis vinifera L.) </title> <p>Vitis vinifera L. has been highlighted by its many positive effects on human health, including antibacterial, anti-inflammatory and antioxidant activity due to its rich phytochemical content. Removing six basal leaves has great potential to influence the production of phytochemicals. The purpose of this study was to determine the impact of different terms of defoliation on the antioxidant and antibacterial activity of grape skin extracts of the Blaufränkisch and Merlot varieties. The total phenolic concentration, total and individual anthocyanin concentration, antioxidant activity and antibacterial activity on gram-positive and gram-negative human pathogens have been determined. The study was conducted on three treatments (control treatment without defoliation, defoliation immediately after bloom and defoliation before the start of the véraison phase). The results showed statistically significant enhancement of the total phenolic concentration as well as the antioxidant and antibacterial activity in both studied cultivars. Defoliation just after blooming was the preferable defoliation term in the Merlot variety for achieving the highest total anthocyanin concentration, antioxidant activity and significant increase of antibacterial activity against all four investigated bacteria. Defoliation before the start of the véraison phase was the preferable defoliation term for achieving the highest total anthocyanin concentration in the Blaufränkisch variety. In general, treatment of defoliation immediately after bloom was more beneficial compared with the defoliation before the start of the véraison phase.</p> <p>Keywords: defoliation; Blaufränkisch; Merlot; phenols; antioxidant activity; antibacterial activity</p> <hd id="AN0137474727-2">1. Introduction</hd> <p>Polyphenols are phytochemicals found extensively in fruits, vegetables and beverages. As secondary plant metabolites, they are commonly involved in defense against ultraviolet radiation or pathogen attack [[<reflink idref="bib1" id="ref1">1</reflink>]]. Epidemiological studies and combined meta-analyses had suggested that extended consummation of diets rich in plant polyphenols give certain protection against the occurrence of cancers, cardiovascular diseases, diabetes, insulin resistance, obesity, neurodegenerative diseases and osteoporosis [[<reflink idref="bib2" id="ref2">2</reflink>]]. The overriding class of biologically active compounds in grapes are polyphenols involving flavonoids, stilbenes and proanthocyanidins. Among the polyphenols found in grapes, flavonoids are the most plentiful biologically-active phytonutrients, possessing cardioprotective, neuroprotective, antimicrobial and antiaging properties [[<reflink idref="bib4" id="ref3">4</reflink>]]. Brightly colored grapes are especially rich in anthocyanins and they have been known for the inhibition of lipid peroxidation and cyclo-oxygenase (COX)-1 and -2 which are inflammatory mediators [[<reflink idref="bib8" id="ref4">8</reflink>]]. The biological activities of flavonoids have been specified to severely hinge on some elements such as the glycosylation degree, sugar residues types and following acyl esterification. Therefore, it is possible to choose different grape cultivars with a distinct flavonoid arrangement having different effects in disease protection [[<reflink idref="bib9" id="ref5">9</reflink>]]. As polyphenols are known for great antioxidant activity, their consumption may ensure some protection against neurological diseases [[<reflink idref="bib10" id="ref6">10</reflink>]]. It was noticed that people who consume three to four glasses of wine per day had an 80% decreased incidence of dementia and Alzheimer's disease compared to those who consumed fewer or none at all [[<reflink idref="bib11" id="ref7">11</reflink>]]. Simple phenolics are also found in grapes and are mostly hydroxycinnamic acid derivatives (<emph>p</emph>-coumaric, caffeic, sinapic and ferulic acids) and hydroxybenzoic acid derivatives (gallic, gentisic, protocatechuic and <emph>p</emph>-hydroxybenzoic acids) [[<reflink idref="bib12" id="ref8">12</reflink>]]. The antibacterial activities of grape, wine and grape-derived products have been broadly considered [[<reflink idref="bib4" id="ref9">4</reflink>], [<reflink idref="bib13" id="ref10">13</reflink>]]. It has been indicated that grape seed extract of "Isabella" grapes var. indicate greater antibacterial activity against gram-positive bacteria in relation to the gram-negative bacteria [[<reflink idref="bib14" id="ref11">14</reflink>]]. Strong pH-dependent inhibitory effects against the growth of <emph>Listeria monocytogenes</emph> have been found for grape juice and skin extracts from black table grapes "Alphonse Lavallée" var., while the seed extract showed a pH-independent inhibitory effect [[<reflink idref="bib15" id="ref12">15</reflink>]]. Muscadine (<emph>Vitis rotundifolia)</emph> grape skin extracts have also been found to be very effective against <emph>H. pylori</emph> in contrast to muscadine grape seed extracts [[<reflink idref="bib16" id="ref13">16</reflink>]].</p> <p>According to recent studies, defoliation has great potential to control microorganisms [[<reflink idref="bib17" id="ref14">17</reflink>]] and influence the production of biocomponents [[<reflink idref="bib18" id="ref15">18</reflink>]]. Defoliation is a powerful and regularly used action to control berry illumination and therefore flavonoid biosynthesis in berries. The obtained effect seems to depend on the defoliation timing and on the genotype [[<reflink idref="bib19" id="ref16">19</reflink>]]. Thus, defoliation can be carried out at different points in time between pre-bloom and <emph>véraison</emph>, but with different outcomes. Early defoliation has shown great potential for efficient control of microorganism contamination, although pre-bloom defoliation leads to reduced grain size and decreased yield of grapes [[<reflink idref="bib17" id="ref17">17</reflink>]]. Grape production and quality are reactive to heat waves, mainly at particular growth stages, such as flowering and ripening [[<reflink idref="bib22" id="ref18">22</reflink>]]. In areas with high rainfall during the winter and high temperatures during the summer season, the defoliation treatment may inhibit ripening and anthocyanin metabolism due to the high temperature of the exposed fruits. Verzera et al. [[<reflink idref="bib23" id="ref19">23</reflink>]] found that the defoliated samples from the later harvest showed the lowest amounts of anthocyanins, which could be due to excessive sunlight exposure of the berries, which causes sunburn damage and does not increase anthocyanin accumulation. Buesa et al. [[<reflink idref="bib24" id="ref20">24</reflink>]] found that post- <emph>véraison</emph> water stress can advance a higher concentration of phenolic substances in berry skins, and in addition, the different response to water availability of grape ripening was found in the Bobal and Tempranillo berries. Severe water stress detrimentally affected berry sugar accumulation in Tempranillo, while the opposite effect was found for Bobal. So, they concluded that the effect of vine water stress depends on the cultivar and on the severity of water stress. Vine water status was the main driver of grape ripening and these responses were genotype-dependent, while ambient temperature appeared to play a minor role in berry ripening. Lanari et al. [[<reflink idref="bib25" id="ref21">25</reflink>]] also found that late leaf removal at <emph>véraison</emph> negatively affected the concentration of anthocyanin and phenolic substances in Montepulciano grapevines, but not in Sangiovese. According to Zhang et al. [[<reflink idref="bib26" id="ref22">26</reflink>]], there is a strait relationship between anthocyanin biosynthesis and berry development. It starts at the <emph>véraison</emph> phase when the biosynthesis of anthocyanins is determined and reaches top level at berry maturity, while at the latest development stages their concentration may be noticeably decreased [[<reflink idref="bib27" id="ref23">27</reflink>]]. However, each grape species and variety has a distinct anthocyanin pattern [[<reflink idref="bib28" id="ref24">28</reflink>]]. The accumulation of tannins and hydroxycinylic acids starts already in the formation of berries and grows until the appearance of <emph>véraison</emph>. For grapes, the highest proportion of phenolic compounds is found in the skin, which is rich in resveratrol (3,5,4′-trihydroxy-stilbene), a polyphenol substance which shows a wide range of biological activities, such as antioxidant and antibacterial activity, and the capability to scavenge free radicals, reduce the risk of cardiovascular disease and also prevent cancer proliferation [[<reflink idref="bib29" id="ref25">29</reflink>]]. Therefore, the main goal of this study was to determine how different terms of ecologically acceptable defoliation of Blaufränkisch and Merlot grape (<emph>Vitis vinifera</emph> L.) leaves affects the total phenolic concentration, total and individual anthocyanin compounds, antioxidant activity and antibacterial activity on gram-positive and gram-negative human pathogens. The study was conducted during 2015 and 2016 on three treatments (control without defoliation, defoliation immediately after bloom and defoliation before the start of the <emph>véraison</emph> phase) in the eastern continental region of Croatia.</p> <hd id="AN0137474727-3">2. Results</hd> <p></p> <hd id="AN0137474727-4">2.1. Effects of Defoliation Treatments on the Concentration of Phenolic Compounds</hd> <p>In the case of defoliation immediately after blooming (T2), the concentration of phenolic compounds was significantly higher (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001) with respect to the control T1 for both grape varieties (Table 1). In the case of the Blaufränkisch variety, the concentration of phenolic compounds was significantly higher in the T2 defoliation treatment compared to the T1 and T3 treatments (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001). Comparison of T2 defoliation treatments of the two investigated varieties showed statistically significant higher concentrations of total phenolic concentration in the Blaufränkisch variety, compared to the Merlot variety (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001). However, the T1 treatments of the two investigated varieties do not differ significantly in the total phenolic concentration. In the case of defoliation before the start of the <emph>véraison</emph> phase (T3) the total phenolic concentration was significantly higher compared to T1 (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001), but less pronounced in relation to T2 (Table 1). This indicates that defoliation immediately after the blooming period is the preferable term for the Blaufränkisch variety for achieving the highest total phenolic concentration. On the other hand, defoliation before the start of the <emph>véraison</emph> phase in the Merlot variety also showed statistically significantly higher concentrations of total phenolic concentration in relation to T1 (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001), but no significant differences between the two defoliation terms were observed. Likewise, the defoliation before the start of the <emph>véraison</emph> phase showed statistically significant higher total phenolic concentrations in the Blaufränkisch variety, compared to the Merlot variety (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001). On the other hand, in the Merlot variety there are significant differences between the two different research years. A generally lower total of phenol concentrations can be noticed during the season 2016.</p> <hd id="AN0137474727-5">2.2. Effects of Defoliation Treatments on the Concentration of Total Anthocyanins</hd> <p>The total concentration of anthocyanins (Table 1), in the case of the Blaufränkisch variety showed that defoliation immediately after blooming in 2015 did not differ significantly in relation to T1. Whereas, in 2016 the T2 treatment resulted in statistically significant lower concentrations of total anthocyanins compared to T1 (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001). On the other hand, in the Merlot variety defoliation immediately after blooming showed statistically significantly higher concentrations of total anthocyanins compared to T1 and T3 (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001). The total concentration of anthocyanins was significantly increased in the case of the Blaufränkisch variety for defoliation before the start of the <emph>véraison</emph> phase compared to the T1 and T2 treatments (U U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001), indicating that the preferable defoliation term for the Blaufränkisch variety for achieving the highest total anthocyanins concentration is defoliation before the start of the <emph>véraison</emph> phase. On the other hand, in the Merlot variety there are no significant differences between T3 and T1, indicating that the preferable defoliation term for the Merlot variety for achieving the highest total anthocyanins concentration is defoliation immediately after blooming.</p> <hd id="AN0137474727-6">2.3. Effects of Defoliation Treatments on the Antioxidant Activity</hd> <p>The differences between the two defoliation times did not prove to be significant in the Blaufränkisch variety for antioxidant activity, but the T3 in the Merlot variety resulted in significantly lower antioxidant activity compared to the T2 (U = 0; N1 = N2 = 9; <emph>p</emph> = 0.006) (Table 1). Thus, both defoliation terms showed statistically significant higher antioxidant activity compared to the T1 (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001). Defoliation before the start of the <emph>véraison</emph> phase showed a statistically significant higher antioxidant activity of the Blaufränkisch variety, compared to the Merlot variety (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001). Statistically significant differences were also observed in comparison of the groups without defoliation of the two examined varieties. The extracts of the Blaufränkisch variety showed significantly higher antioxidant activity compared to the Merlot varieties T1 extracts (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001).</p> <p>It was found that the extracts obtained after defoliation treatments, irrespective of the defoliation time, had significantly higher antioxidant activity than without defoliation. The differences between the two defoliation times did not prove to be significant in the Blaufränkisch variety, but T2 resulted in significantly higher antioxidant activity compared to the T1 and T3 in the Merlot variety (U = 0; N1 = N2 = 9; <emph>p</emph> = 0.006). It could be concluded that the defoliation immediately after blooming is the preferable defoliation term for gaining better antioxidant activity of the Merlot variety extracts. Comparison of the two cultivars investigated did not show statistically significant differences in the antioxidant activity of the Blaufränkisch variety extracts relative to the Merlot variety in the case of defoliation immediately after blooming.</p> <hd id="AN0137474727-7">2.4. Effects Of Defoliation Treatments on Individual Anthocyanins</hd> <p>Quantitative analysis of individual anthocyanins based on high-performance liquid chromatography in the Blaufränkisch variety extracts in 2016 is presented in Table 2. Blaufränkisch extracts from 2015 were not analysed. Among the identified anthocyanins, eight are significantly decreased by defoliation immediately after blooming in 2016: delphinidin-3-glucoside, petunidin-3-glucoside, malvidin-3-glucoside, peonidin-3-(acetyl)glucoside, malvidin-3-(acetyl)glucoside, cyanidin-3-<emph>O</emph>-(6″-<emph>p</emph>-(coumaroyl)-glucoside, peonidin-3-<emph>O</emph>-(6-<emph>p</emph>-(coumaroyl)-glucoside and malvidin-3-<emph>O</emph>-(6-<emph>p</emph>-(coumaroyl)-glucoside (trans isomer). Defoliation before the start of the <emph>véraison</emph> phase did not prove to be significant in the Blaufränkisch variety extracts in 2016 (Table 2).</p> <p>Defoliation immediately after blooming in the case of Merlot variety did not result in significantly different concentrations in 2015, but in 2016 resulted in significantly increased concentrations of individual anthocyanins (Table 3). Every identified anthocyanin significantly increased in concentration in 2016: delphinidin-3-glucoside, cyanidin-3-glucoside, petunidin-3-glucoside, peonidin-3-glucoside, malvidin-3-glucoside, delphinidin-3-(acetyl)-glucoside, cyanidin-3-(acetyl)-glucoside, petunidin-3-(acetyl)-glucoside, delphinidin-3-<emph>O</emph>-(6-<emph>p</emph>-(coumaroyl)-glucoside, peonidin-3-(acetyl) glucoside, malvidin-3-(acetyl) glucoside, cyanidin-3-<emph>O</emph>-(6″-<emph>p</emph>-(coumaroyl)-glucoside, petunidin-3-<emph>O</emph>-(6-<emph>p</emph>-(coumaroyl)-glucoside, malvidin-3-<emph>O</emph>-(6-<emph>p</emph>-(coumaroyl)-glucoside (cis isomer), peonidin-3-<emph>O</emph>-(6-<emph>p</emph>-(coumaroyl)-glucoside and malvidin-3-<emph>O</emph>-(6-<emph>p</emph>-(coumaroyl)-glucoside (trans isomer).</p> <p>Defoliation before the start of the <emph>véraison</emph> phase in the case of the Merlot variety resulted in significantly decreased concentrations of individual antocyanins in 2015 compared to T2 and T1, but in 2016 did not prove to be significant (Table 3), as with the Blaufränkisch variety extracts in 2016. Among the identified anthocyanins, nine of them significantly decreased concentration in 2015: delphinidin-3-glucoside, petunidin-3-glucoside, malvidin-3-glucoside, delphinidin-3-<emph>O</emph>-(6-<emph>p</emph>-(coumaroyl)-glucoside, peonidin-3-(acetyl) glucoside, malvidin-3-(acetyl) glucoside, petunidin-3-<emph>O</emph>-(6<emph>-p</emph>-(coumaroyl)-glucoside, malvidin-3-<emph>O</emph>-(6-<emph>p</emph>-(coumaroyl)-glucoside (<emph>cis</emph> isomer) and malvidin-3-<emph>O</emph>-(6-<emph>p</emph>-(coumaroyl)-glucoside (<emph>trans</emph> isomer).</p> <hd id="AN0137474727-8">2.5. Effects of Defoliation Treatments on Antibacterial Activity</hd> <p>The best effect in gaining stronger antibacterial activity of Blaufränkisch and Merlot grape skin extracts was obtained by defoliation immediately after blooming in the Blaufränkisch variety, as it resulted in a significant decrease of MIC values in all four investigated bacteria (Table 4). In general, the MIC values where higher in 2016, but it can be seen that the MIC values were almost two times higher in 2016 for <emph>P. aeruginosa</emph> and <emph>S. aureus</emph> in the Blaufränkisch variety, and for <emph>S. aureus</emph> in the Merlot variety. In the case of the Blaufränkisch variety, defoliation immediately after blooming showed statistically significantly stronger antibacterial activity compared to the defoliation before the start of <emph>véraison</emph>, and also compared to the group without defoliation. On the other hand, in the Merlot variety defoliation immediately after blooming showed statistically significant stronger antibacterial activity compared to the group without defoliation (U = 0; N1 = N2 = 9; <emph>p</emph> &lt; 0.001), but not compared to the defoliation before the start of <emph>véraison</emph>, except for <emph>P. aeruginosa</emph> and <emph>S. aureus</emph>. Defoliation before the start of <emph>véraison</emph> resulted in a significant decrease of MIC values in all four investigated bacteria. On the other hand, in the Merlot variety defoliation before the start of <emph>véraison</emph> did not prove to be significant.</p> <hd id="AN0137474727-9">3. Discussion</hd> <p>Our previous work [[<reflink idref="bib32" id="ref26">32</reflink>]] showed that early leaf removal treatments resulted in statistically significant increased total phenolic concentration, total anthocyanin compounds, and antioxidant activity measured in extracts of grape skin. Therefore we performed ecologically acceptable defoliation of Blaufränkisch and Merlot grape (<emph>Vitis vinifera</emph> L.) leaves to investigate the effect of two different times of defoliation: immediately after bloom and before the start of the <emph>véraison</emph> phase. The timing of partial defoliation is proven to affect the grape vegetative response. The research of Hunter et al. [[<reflink idref="bib22" id="ref27">22</reflink>]] confirmed that the earlier the defoliation was applied, the more the lateral shoot length and the number of lateral shoots increased, resulting in a higher total shoot length. While if defoliation was performed at <emph>véraison</emph> it had no effect on lateral growth. In our research, defoliation immediately after blooming increased the total phenolic concentration for both grape varieties, achieved the highest total anthocyanins concentration and antioxidative activity in the case of the Merlot variety, and achieved the best effect in gaining stronger antibacterial activity of the Blaufränkisch and Merlot grape skin extracts. The relationship between berry skin composition and defoliation seems to be cultivar dependent. The increased total phenolic concentration after the defoliation treatment performed prior to blooming was noted by Poni et al. [[<reflink idref="bib33" id="ref28">33</reflink>]]. According to research carried out by Risco et al. [[<reflink idref="bib34" id="ref29">34</reflink>]] early defoliation of the <emph>Vitis vinifera</emph> L., Nero d'Avola variety resulted in an increase of the total content of anthocyanin, flavonoids and polyphenols [[<reflink idref="bib23" id="ref30">23</reflink>]]. Thus, results also showed that overall vine performance was not stimulated by the defoliation treatments. Mijowska et al. [[<reflink idref="bib35" id="ref31">35</reflink>]] found that early defoliation, particularly pre-flowering, resulted in a significant increase in total polyphenol content. Results from the research of Jerman et al. [[<reflink idref="bib36" id="ref32">36</reflink>]] on grape varieties of Pinot Noir indicate that early defoliation increases antioxidant activity. The research carried out by Radovanović et al. (2015) showed that the early defoliation of the Cabernet Sauvignon variety resulted in a significant increase in total phenol concentrations of up to 88.75%, as well as an increase in antioxidant activity up to 12.70% compared to the control group. The total concentration of anthocyanins in the case of the Blaufränkisch variety showed different trends between two different research years. Defoliation immediately after blooming in 2015 did not differ significantly in relation to the control group, but in 2016 the treatment resulted in statistically significant lower concentrations of total anthocyanins. Also, a generally lower total phenol concentration can be noticed during 2016 in the Merlot variety. This is in accordance with previous research, that the weather conditions have a considerable impact on the amounts of antioxidant components in grape skin extracts [[<reflink idref="bib32" id="ref33">32</reflink>]]. As it can be seen from the Table 5 and Figure 1 because the cumulative rainfall was much higher, and min-max temperatures were lower for 2016, it could be concluded that precipitation is an important climatic factor in terms of the formation of phenolic compounds in the skin of grapes [[<reflink idref="bib37" id="ref34">37</reflink>]]. The increase in the number of days with warm temperatures is particularly relevant for vineyards also. According to Ferrer et al. [[<reflink idref="bib38" id="ref35">38</reflink>]] and Barbagallo et al. [[<reflink idref="bib39" id="ref36">39</reflink>]], the concentration of individual anthocyanins has shown the utmost alteration between the years. Our results of decreased concentration of anthocyanins with excess rainfall during the vegetation period in 2016 are in accordance with results of Ferrer et al. [[<reflink idref="bib38" id="ref37">38</reflink>]] and Roby et al. [[<reflink idref="bib40" id="ref38">40</reflink>]] where water deficits increased the amount of skin anthocyanins. Berry temperature is important, as it is affected not only by sunlight exposure but also by the availability of water to maintain transpiration. Sunlight exposure, combined with simultaneous high temperatures, leads to a decrease in phenolic compounds [[<reflink idref="bib6" id="ref39">6</reflink>]]. Jerman et al. [[<reflink idref="bib36" id="ref40">36</reflink>]] followed an air temperature/precipitation behaviour in addition to a grape surface temperature monitoring during an experimental trial of early leaf removal of Pinot Noir' grapes from Vipava Valley. The temperatures of the grapes were consistently higher (4 °C on average) in comparison to atmospheric ones, but apparently not affected the synthesis of phenols, since the increases of total anthocyanins and polyphenols till the mid of August were observed. Interestingly, their concentrations have decreased afterwards, most likely due to a precipitation-related grape phenols dilution effect. All these environmental factors can have an effect on the development of vines and their particular phenological stages, with consequences on berry quality, but the precise response is dependent on the grapevine genotype [[<reflink idref="bib19" id="ref41">19</reflink>]]. Therefore, it appears that the response to leaf removal may vary depending on the cultivar. In our research, defoliation before the start of <emph>véraison</emph> resulted in a significant increase in total anthocyanins concentration and antioxidative activity in the case of the Blaufränkisch variety, in contrast to the Merlot variety. This also resulted in a significant decrease of MIC values in all four investigated bacteria, but less pronounced compared to the defoliation immediately after blooming. However, the best antibacterial activity in general (Table 4) was found with the Merlot variety after defoliation immediately after blooming in 2015, despite the lower determined values of TAC and TPC compared to Blaufränkisch. This may be due to a greater number of individual anthocyanin components in Merlot or complex mixtures of plant metabolites, such as alkaloids, saponins, tannins, amino acids, triterpenoids, phlobatannins, steroids and catechol found in grape skin [[<reflink idref="bib41" id="ref42">41</reflink>]]. Secondary metabolite distribution may also be modified during plant growth, or linked to the changes of climatic conditions, so in future studies it would be of great interest to perform phytochemical screening. The defoliation performed by Intrigliolo et al. [[<reflink idref="bib42" id="ref43">42</reflink>]], at the full maturity of grapes on the Mandó cultivar from Southeast Spain resulted in a reduction in grain size, but also increased concentrations of phenol, anthocyanin and tannins in grape berries. In red grape varieties anthocyanin accumulation starts from <emph>véraison</emph> and achieves its limits in the finite phases of fruit maturation when the synthesis runs down [[<reflink idref="bib43" id="ref44">43</reflink>]], but phenolic synthesis and accumulation in grape berry is also determined by genetic factors and the interaction between genotype and environment [[<reflink idref="bib44" id="ref45">44</reflink>]]. It can be concluded that the defoliation treatment results in an increased phenolic concentration in the grape skin, but whether different terms of leaf removal will result in different increases in phenolic concentration is highly specific for each individual grape variety. Given that no previous research has been carried out thus far to describe the effect of defoliation treatment on the antibacterial activity of grape skin extracts, this work is unique. Previous studies have investigated the antibacterial activity of grape skin extracts but without considering the effect of defoliation treatment, especially in different terms. Nirmala and Narendhirakannan [[<reflink idref="bib41" id="ref46">41</reflink>]] found that <emph>Vitis vinifera</emph> L. Muscat from the Coimbatore district of Tamil Nadu, India are rich in antioxidants and possess antimicrobial activity. Katalinić et al. [[<reflink idref="bib46" id="ref47">46</reflink>]] conducted a study in which they studied the antibacterial effect of grape skin extracts of 14 <emph>V. vinifera</emph> varieties grown in Dalmatia on gram-positive and gram-negative bacteria. Lower MIC values were confirmed for phenolic mixtures from white grape cultivars for all tested species which was contrary to expectations. The investigated grape skin extracts showed good antibacterial as well as antioxidant activity, which are of paramount importance when it comes to human health. The positive effects of grapes in the form of prevention of the occurrence of certain diseases, or at least their progression, are very interesting and relevant for research. It would be desirable to compare different extraction methods with solvents of different polarity or supercritical fluid extraction, combined with the effect of various new food technologies such as ultrasound, high hydrostatic pressure and pulsating electric field treatments.</p> <hd id="AN0137474727-10">4. Materials and Methods</hd> <p></p> <hd id="AN0137474727-11">4.1. Plant Material and Experimental Design</hd> <p>Two <emph>V. vinifera</emph> varieties, Blaufränkisch (VIVC-Variety-No.: 1459) and Merlot Noir (VIVC-Variety-No.: 7657), were studied during 2015 and 2016. A vineyard, planted in 2013, is situated at the transition from the luvisol to the stagnic luvisol, located in Mandićevac (lat. 45. 368 417, lon. 18. 245 611, elevation 208 m), in the eastern continental region of Croatia, the subregion of Slavonija, among the vineyards of Đakovo, with south exposure and a general fall W→E of 9.8%. The chemical properties of this soil indicate an acid reaction. The vine training system was single Guyot, with 12 buds per plant. The vines were planted with 2.2 m spacing between rows and 0.8 m within rows, for a total of 5681 vines/ha. The experiment was designed by random block formation consisting of three replicates. The study was conducted on three treatments: control (T1), and two different treatments of six basal leaf removal were performed: defoliation immediately after bloom: BBCH 69 phase (T2) and defoliation before the start of <emph>véraison</emph> was performed during BBCH 79 phase (T3) according to the extended BBCH scale [[<reflink idref="bib47" id="ref48">47</reflink>]]. The treatments included ten plants per every separate treatment in three repetitions, which was 90 vines in total. The experimental field did not supply an irrigation system. Rainfall and mean temperatures for 2015 and 2016 during the March–October time period were obtained from the Meteorological and Hydrological Service of Croatia and presented in Table 5. Cumulative rainfall during the April–September period in 2016 was higher than in 2015, while the mean temperatures were equivalent. The trend of mean temperatures from the T3 treatment till harvest date can be seen in the Figure 1a,b, for 2015 and 2016, respectively, while mean min-max temperatures for both years from the <emph>véraison</emph> phase till harvest date can be seen in the Figure 1c.</p> <hd id="AN0137474727-12">4.2. Grape Skin Extraction</hd> <p>Grape skin extraction was performed according to the method described by Rustioni et al. [[<reflink idref="bib48" id="ref49">48</reflink>]]. Each sample represents more randomly selected clusters from which ten berries were separated. The grape skin was manually detached from the pulp and samples were gently dried with a napkin to avoid loss of anthocyanins. Thereafter, the mixture was soaked for 20 h with 20 mL of a mixture of ethanol: water: hydrochloric acid (70:29:1, <emph>w/w/w</emph>). Each sample represents more randomly selected clusters from which 10 berries were separated in each repetition. The total phenolic concentration from the extract was determined after filtration. The extraction procedure was carried out with three biological replicates.</p> <hd id="AN0137474727-13">4.3. Determination of Total Phenolic Concentration (TPC)</hd> <p>The total phenolic concentration of grape skin extracts was determined by a spectrophotometric method which used the Folin–Ciocalteu reagent. The standard calibration (0.018–0.50 mg mL<sups>−1</sups>) curve was plotted using gallic acid [[<reflink idref="bib49" id="ref50">49</reflink>]]. The results were derived from triplicate analyses, normalized against negative control of solvent and expressed as milligrams of gallic acid equivalents (GAE) per gram of grape skin and reported as mean ± standard deviation (SD).</p> <hd id="AN0137474727-14">4.4. Determination of Total Anthocyanins (TAC)</hd> <p>The total anthocyanins were determined by a spectrophotometric method described by Nagel and Wulf [[<reflink idref="bib50" id="ref51">50</reflink>]]. Absorbance of each dilution was measured at the 540 nm, against a blank with distilled water. The total anthocyanin concentration results were derived from triplicate analyses, normalized against negative control of solvent and expressed in mg of malvidin-3-<emph>O</emph>-glucoside equivalents (MAE) per g of grape skin, using the molar extinction coefficient (ε) of malvidin-3-<emph>O</emph>-glucoside of 28,000 L mol<sups>−1</sups> cm<sups>−1</sups> and molar weight (MW) (493.2 g mol<sups>−1</sups>) and reported as mean ± standard deviation (SD).</p> <hd id="AN0137474727-15">4.5. 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) Radical Scavenging Activity</hd> <p>The total antioxidant activities of grape skin extracts were determined using the DPPH radical scavenging assay described earlier [[<reflink idref="bib51" id="ref52">51</reflink>]]; 750 μL of the diluted grape skin extracts (average final concentration 100 mg mL<sups>−1</sups> for Merlot variety and 150 mg mL<sups>−1</sups> for Blaufränkisch variety) was mixed with the same amount of 0.2 mM DPPH radical solution, so the final DPPH radical concentration was 0.1 mM. The mixture was well stirred and incubated at room temperature for 30 min. Ascorbic acid (AA) was used as a reference compound in the concentration range 2–200 μg mL<sups>−1</sups>. All experiments were performed in triplicate. The absorbance decrease at 517 nm was measured, and DPPH scavenging activity was established using Equation (<reflink idref="bib1" id="ref53">1</reflink>):</p> <p>DPPH activity = (A<subs>b</subs> + A<subs>s</subs>) − A<subs>m</subs>)/A<subs>b</subs> × 100 (<reflink idref="bib1" id="ref54">1</reflink>)</p> <p>where A<subs>b</subs> is the absorbance of 0.1 mM DPPH radical solution at λ = 517 nm, A<subs>s</subs> is the absorbance of 0.1 mM extraction solution at λ = 517 nm, and A<subs>m</subs> is the absorbance of 0.1 mM solution mixture of tested extracts and DPPH radical at 517 nm.</p> <hd id="AN0137474727-16">4.6. High-Performance Liquid Chromatography (HPLC): Separation and Detection of Anthocyanins</hd> <p>The anthocyanins were analysed according to OIV-MA-AS315-11 with some modifications [[<reflink idref="bib52" id="ref55">52</reflink>]]. Anthocyanins were analysed by direct injection of the samples, previously centrifuged and filtered through a 0.45 μm pore size membrane filter, in an Agilent 1100/1200 series HPLC system equipped with an Agilent photodiode array detector (Agilent Technologies, Wilmington, NC, USA). The separation was performed on a reversed phase column: LiChrospher 100 RP 18 (5 µm) in LiChroCart 250–4 with guard column LiChroCart 4 mm RP 18 all obtained from Merck (Darmstadt, Germany). Separation was performed with solvents: (A) water/formic acid/acetonitrile (87:10:3, <emph>v/v/v</emph>) and (B) water/formic acid/acetonitrile (40:10:50, <emph>v/v/v</emph>). The following gradient of eluents was used: 6%–30% (B) 0–15 min, 30%–50% (B) 15–30 min, 50%–60% (B) 30–35 min, 60%–6% (b) 35–41 min. The flow rate was 0.4 mL min<sups>−1</sups>, detection wavelength 520 nm and the analysis were performed at 20 °C. Individual anthocyanins were identified by comparing their retention times with external standards and literature [[<reflink idref="bib53" id="ref56">53</reflink>]]. Diglycosides were detected in very low and not relevant amounts and they were excluded accordingly. Available standard substances of cyanidin-3,5- diglucoside, cyanidin-3-glucoside, delphinidin-3-glucoside, malvidin-3,5-diglucoside, petunidin- 3-glucoside, peonidin-3-glucoside, malvidin-3-glucoside, all obtained from Sigma-Aldrich (St. Louis, MO, USA). Anthocyanins were quantified by using a seven-point external calibration curve (R<sups>2</sups> = 0.9997) obtained by injecting standard solutions of malvidin-3-monoglucoside chloride. Results were expressed as malvidine-3-glucoside equivalents MAE per gram of grape skin. All analyses were done in triplicate and results expressed as mean ± standard deviation (SD).</p> <hd id="AN0137474727-17">4.7. Antibacterial Susceptibility Testing</hd> <p></p> <hd id="AN0137474727-18">4.7.1. Microorganisms and Growth Conditions</hd> <p> <emph>Bacillus subtilis</emph> and <emph>Staphylococcus aureus</emph> as two gram-positive, and <emph>Escherichia coli</emph> and <emph>Pseudomonas aeruginosa</emph> as gram-negative bacterial strains, were used to determine the antibacterial activities of the grape skin extracts. These bacteria were isolates from various clinical specimens obtained from the Microbiology Service of the Public Health Institute of Osijek–Baranja County (Osijek, Croatia). Working cultures were prepared from subcultures and grown overnight in Muller Hinton Broth (MHB) (Fluka Analytical, Buchs, Switzerland) under optimal conditions (37 °C with 50% humidity). The antibacterial standard Amikacin sulfate (Biorisan, Amikacin, Vocate, Glyfada, Greece) was dissolved in sterile distilled water.</p> <hd id="AN0137474727-19">4.7.2. Minimum Inhibitory Concentrations</hd> <p>MIC values were determined by a modified microdilution method [[<reflink idref="bib57" id="ref57">57</reflink>]] as described in our previous work [[<reflink idref="bib58" id="ref58">58</reflink>]]. Briefly, a total of 100 μL of midlogarithmic-phase bacterial cultures (5 × 10<sups>5</sups> CFU mL<sups>−1</sups>) in Mueller Hinton Broth were added to 100 μL of two-fold serially diluted extracts (250–0.122 mg mL<sups>−1</sups>). Wells containing bacterial inoculum without extracts (growth control) and wells containing only broth and solvent (background control) were included in each plate. The antibacterial standard amikacin sulfate was co-assayed under the same conditions. The MIC value was defined as the lowest concentrations of extract at which there was no color change or visual turbidity due to microbial growth, derived from triplicate analyses, normalized against negative control and expressed as micrograms per milliliter.</p> <hd id="AN0137474727-20">4.8. Statistical Data Processing</hd> <p>All statistical analysis were achieved by the statistical program STATISTICA 12.0 (Statsoft, Inc., Tulsa, OK, USA). The normality of the distribution of numeric variables was tested by the Shapiro–Wilk test. Given that the data does not follow normal distribution, the nonparametric Mann–Whitney U-test was used. All tests were performed at a level of significance of α = 0.05.</p> <hd id="AN0137474727-21">5. Conclusions</hd> <p>Both defoliation treatments significantly increased the total phenolic concentration as well as the antioxidant and antibacterial activity of the grape skin extracts in both examined <emph>Vitis vinifera</emph> L. varieties Blaufränkisch and Merlot. Defoliation just after blooming is the preferable defoliation term in the Merlot variety for achieving the highest total anthocyanin concentration, gaining the highest antioxidant activity and a significant increase of antibacterial activity against all four investigated bacteria. Defoliation before the start of the <emph>véraison</emph> phase is the preferable defoliation term for achieving the highest total anthocyanin concentration in the Blaufränkisch variety. In general, the treatment of defoliation immediately after bloom was more successful under the given experimental conditions compared with the defoliation before the start of the <emph>véraison</emph> phase.</p> <hd id="AN0137474727-22">Figure and Tables</hd> <p>Graph: Figure 1 (a) Mean temperature (°C, left axis) and rainfall (mm, right axis) trend from T3 treatment till harvest date in 2015; (b) Mean temperature and rainfall trend from T3 treatment till harvest in 2016; (c) Mean with min-max mean temperatures from véraison till harvest date.</p> <p>Table 1 Total phenolic concentration (TPC) in grape skin extracts expressed as mg of GAE per g of grape skin, Total anthocyanins concentration (TAC) in grape skin extracts expressed as mg of MAE per g of grape skin and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity expressed as % DPPH radical scavenging activity of Blaufränkisch and Merlot variety.</p> <p> <ephtml> &lt;table&gt;&lt;thead&gt;&lt;tr&gt;&lt;th align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;&lt;italic&gt;Vitis vinifera&lt;/italic&gt; L. variety&lt;/th&gt;&lt;th align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Year &lt;/th&gt;&lt;th align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Treatment&lt;/th&gt;&lt;th align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;TPC (mgGAE g&lt;sub&gt;skin&lt;/sub&gt;&lt;sup&gt;&amp;#8722;1&lt;/sup&gt;)&lt;/th&gt;&lt;th align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;TAC (mgMAE g&lt;sub&gt;skin&lt;/sub&gt;&lt;sup&gt;&amp;#8722;1&lt;/sup&gt;)&lt;/th&gt;&lt;th align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;DPPH Radical Scavenging Activity (%)&lt;/th&gt;&lt;/tr&gt;&lt;/thead&gt;&lt;tbody&gt;&lt;tr&gt;&lt;td rowspan="6" align="center" valign="middle" style="border-bottom:solid thin"&gt;Blaufr&amp;#228;nkisch &lt;/td&gt;&lt;td rowspan="3" align="center" valign="middle"&gt;2015&lt;/td&gt;&lt;td align="center" valign="middle"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.79 &amp;#177; 0.59&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;2.06 &amp;#177; 0.16&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;54.03 &amp;#177; 0.53&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;18.2 &amp;#177; 0.06&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;1.93 &amp;#177; 0.11&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;56.52 &amp;#177; 0.91&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;T3&lt;/td&gt;&lt;td align="center" valign="middle"&gt;17.26 &amp;#177; 0.53&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;2.37 &amp;#177; 0.09&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;55.86 &amp;#177; 0.31&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td rowspan="3" align="center" valign="middle" style="border-bottom:solid thin"&gt;2016&lt;/td&gt;&lt;td align="center" valign="middle"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.86 &amp;#177; 0.26&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;1.98 &amp;#177; 0.09&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;56.34 &amp;#177; 0.84&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;17.12 &amp;#177; 1.02&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;1.72 &amp;#177; 0.03&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;57.69 &amp;#177; 0.85&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T3&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;15.61 &amp;#177; 0.18&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;2.35 &amp;#177; 0.11&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;57.06 &amp;#177; 0.08&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td rowspan="6" align="center" valign="middle" style="border-bottom:solid thin"&gt;Merlot &lt;/td&gt;&lt;td rowspan="3" align="center" valign="middle"&gt;2015&lt;/td&gt;&lt;td align="center" valign="middle"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.85 &amp;#177; 0.22&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;1.60 &amp;#177; 0.06&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;51.97 &amp;#177; 0.34&lt;sub&gt; b &lt;/sub&gt; B &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;14.97 &amp;#177; 0.31&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;1.84 &amp;#177; 0.04&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;57.18 &amp;#177; 0.49&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;T3&lt;/td&gt;&lt;td align="center" valign="middle"&gt;14.96 &amp;#177; 0.63&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;1.62 &amp;#177; 0.05&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;54.04 &amp;#177; 0.97&lt;sub&gt; b &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td rowspan="3" align="center" valign="middle" style="border-bottom:solid thin"&gt;2016&lt;/td&gt;&lt;td align="center" valign="middle"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.73 &amp;#177; 0.33&lt;sub&gt; b &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;1.02 &amp;#177; 0.07&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;52.86 &amp;#177; 1.55&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;9.29 &amp;#177; 0.27&lt;sub&gt; b &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;1.32 &amp;#177; 0.04&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;57.50 &amp;#177; 0.63&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T3&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;8.36 &amp;#177; 0.13&lt;sub&gt; b &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;1.09 &amp;#177; 0.12&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;56.59 &amp;#177; 0.30&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;/tbody&gt;&lt;/table&gt; </ephtml> Different green subscript lower case letters in each row indicate statistically significant differences (<emph>p</emph>&lt; 0.001) between years for the same variety and same treatment. Different blue upper case letters in each row indicate statistically significant differences (<emph>p</emph> &lt; 0.001) between two varieties for the same year and same treatment. Different magenta superscript lower case letters in each row indicate statistically significant differences (<emph>p</emph> &lt; 0.001) between treatments for the same variety and same year.</p> <p>Table 2 Individual anthocyanins concentration in grape skin extracts expressed as mg of MAE per g of grape skin for Blaufränkisch variety.</p> <p> <ephtml> &lt;table&gt;&lt;thead&gt;&lt;tr&gt;&lt;th align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Skin anthocyanins (mg&lt;sub&gt;MAE&lt;/sub&gt; g &lt;sub&gt;skin&lt;/sub&gt;&lt;sup&gt;&amp;#8722;1&lt;/sup&gt;)&lt;/th&gt;&lt;th colspan="6" align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Blaufr&amp;#228;nkisch&lt;/th&gt;&lt;/tr&gt;&lt;/thead&gt;&lt;tbody&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;Year&lt;/td&gt;&lt;td colspan="3" align="center" valign="middle" style="border-bottom:solid thin"&gt;2015&lt;/td&gt;&lt;td colspan="3" align="center" valign="middle" style="border-bottom:solid thin"&gt;2016&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;Treatment&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T3&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T3&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Delphinidin-3-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0665 &amp;#177; 0.0102 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0498 &amp;#177; 0.0058 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.059 &amp;#177; 0.0095 &lt;sup&gt;a,b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Cyanidin-3-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0312 &amp;#177; 0.0107 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0337 &amp;#177; 0.0095 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0403 &amp;#177; 0.0101 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Petunidin-3-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0867 &amp;#177; 0.0069 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0605 &amp;#177; 0.0048 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0762 &amp;#177; 0.0071 &lt;sup&gt;c&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Peonidin-3-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.3372 &amp;#177; 0.0585 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.3098 &amp;#177; 0.0656 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.3961 &amp;#177; 0.0587 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Malvidin-3-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.9323 &amp;#177; 0.0547 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.5611 &amp;#177; 0.0459 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.8167 &amp;#177; 0.1464 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Petunidin-3-(acetyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0005 &amp;#177; 0.0004 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0005 &amp;#177; 0.000 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0009 &amp;#177; 0.0004 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Delphinidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6-&lt;italic&gt;p&lt;/italic&gt;-(coumaroyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0028 &amp;#177; 0.0013 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0022 &amp;#177; 0.0001 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0031 &amp;#177; 0.0005 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Peonidin-3-(acetyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0021 &amp;#177; 0.0002 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0016 &amp;#177; 0.0003 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0023 &amp;#177; 0.0005 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Malvidin-3-(acetyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0120 &amp;#177; 0.0013 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0059 &amp;#177; 0.0012 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0110 &amp;#177; 0.0036 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Cyanidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6&amp;#8243;-&lt;italic&gt;p&lt;/italic&gt;-(coumaroyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0035 &amp;#177; 0.0003 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0025 &amp;#177; 0.0004 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0039 &amp;#177; 0.0006 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Petunidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6&lt;italic&gt;-p&lt;/italic&gt;-(coumaroyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0041 &amp;#177; 0.0023 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0034 &amp;#177; 0.0001 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0047 &amp;#177; 0.0006 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Malvidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6-&lt;italic&gt;p&lt;/italic&gt;-(coumaroyl)-glucoside (&lt;italic&gt;cis&lt;/italic&gt; isomer)&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0029 &amp;#177; 0.0018 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0016 &amp;#177; 0.0001 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0027 &amp;#177; 0.0008 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Peonidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6-&lt;italic&gt;p&lt;/italic&gt;-(coumaroyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0252 &amp;#177; 0.0005 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0165 &amp;#177; 0.001 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0221 &amp;#177; 0.0039 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Malvidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6&lt;italic&gt;-p&lt;/italic&gt;-(coumaroyl)-glucoside (&lt;italic&gt;trans&lt;/italic&gt; isomer)&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.073 &amp;#177; 0.0108 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0377 &amp;#177; 0.0104 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom"&gt;0.0638 &amp;#177; 0.0229 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;&amp;#8721; c Anthocyanins (with unknown)&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;n.d.&lt;/td&gt;&lt;td align="center" valign="bottom" style="border-bottom:solid thin"&gt;1.5908 &amp;#177; 0.0237 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom" style="border-bottom:solid thin"&gt;1.0944 &amp;#177; 0.0357 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="bottom" style="border-bottom:solid thin"&gt;1.5144 &amp;#177; 0.109 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;/tbody&gt;&lt;/table&gt; </ephtml> Different superscript lower case letters in each row indicate statistically significant differences (<emph>p</emph> &lt; 0.001) between treatments.</p> <p>Table 3 Individual anthocyanins concentration in grape skin extracts expressed as mg of MAE per g of grape skin for Merlot variety.</p> <p> <ephtml> &lt;table&gt;&lt;thead&gt;&lt;tr&gt;&lt;th align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Skin anthocyanins (mg&lt;sub&gt;MAE&lt;/sub&gt; g &lt;sub&gt;skin&lt;/sub&gt;&lt;sup&gt;&amp;#8722;1&lt;/sup&gt;)&lt;/th&gt;&lt;th colspan="6" align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Merlot&lt;/th&gt;&lt;/tr&gt;&lt;/thead&gt;&lt;tbody&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;Year&lt;/td&gt;&lt;td colspan="3" align="center" valign="middle" style="border-bottom:solid thin"&gt;2015&lt;/td&gt;&lt;td colspan="3" align="center" valign="middle" style="border-bottom:solid thin"&gt;2016&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;Treatment&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T3&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;T3&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Delphinidin-3-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.1384 &amp;#177; 0.0186 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.1382 &amp;#177; 0.0022 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.1247 &amp;#177; 0.0004 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0424 &amp;#177; 0.0023 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0633 &amp;#177; 0.0014 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0397 &amp;#177; 0.0114 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Cyanidin-3-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0207 &amp;#177; 0.0032 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0202 &amp;#177; 0.0048 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0201 &amp;#177; 0.0043 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0115 &amp;#177; 0.0014 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0184 &amp;#177; 0.0035 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0140 &amp;#177; 0.0026 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Petunidin-3-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.1347 &amp;#177; 0.0134 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.1353 &amp;#177; 0.0032 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.1168 &amp;#177; 0.0067 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.043 &amp;#177; 0.0016 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0674 &amp;#177; 0.0093 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0422 &amp;#177; 0.008 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Peonidin-3-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0687 &amp;#177; 0.0138 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.072 &amp;#177; 0.0143 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0611 &amp;#177; 0.0031 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0457 &amp;#177; 0.008 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0753 &amp;#177; 0.0087 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0534 &amp;#177; 0.0066 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Malvidin-3-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.5212 &amp;#177; 0.0436 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.5293 &amp;#177; 0.0143 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.4169 &amp;#177; 0.0507 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.2048 &amp;#177; 0.0113 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.3318 &amp;#177; 0.0482 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.1948 &amp;#177; 0.0197 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Delphinidin-3-(acetyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0201 &amp;#177; 0.0029 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.021 &amp;#177; 0.0023 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0186 &amp;#177; 0.0026 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0051 &amp;#177; 0.0013 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0079 &amp;#177; 0.0014 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0048 &amp;#177; 0.0013 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Cyanidin-3-(acetyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.003 &amp;#177; 0.0004 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0036 &amp;#177; 0.0009 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0032 &amp;#177; 0.001 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0008 &amp;#177; 0.0001 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0015 &amp;#177; 0.0004 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0009 &amp;#177; 0.0005 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Petunidin-3-(acetyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0207 &amp;#177; 0.0026 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0207 &amp;#177; 0.0018&lt;sup&gt;a,b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0178 &amp;#177; 0.003 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0051 &amp;#177; 0.0019 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0086 &amp;#177; 0.0008 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0052 &amp;#177; 0.0008 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Delphinidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6-&lt;italic&gt;p&lt;/italic&gt;-(coumaroyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0305 &amp;#177; 0.0024 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0301 &amp;#177; 0.0015 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0251 &amp;#177; 0.004 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0067 &amp;#177; 0.0002 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0104 &amp;#177; 0.001 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0062 &amp;#177; 0.0008 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Peonidin-3-(acetyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0085 &amp;#177; 0.0022 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0084 &amp;#177; 0.0031 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0048 &amp;#177; 0.0031 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0047 &amp;#177; 0.0006 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0065 &amp;#177; 0.0025 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0041 &amp;#177; 0.0016 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Malvidin-3-(acetyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.1077 &amp;#177; 0.0123 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.093 &amp;#177; 0.0322 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0513 &amp;#177; 0.033 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0338 &amp;#177; 0.0028 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0558 &amp;#177; 0.0072 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0298 &amp;#177; 0.0035 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Cyanidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6&amp;#8243;-&lt;italic&gt;p&lt;/italic&gt;-(coumaroyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0095 &amp;#177; 0.0013 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.015 &amp;#177; 0.0053 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0127 &amp;#177; 0.0073 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0052 &amp;#177; 0.0008 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0081 &amp;#177; 0.0009 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0052 &amp;#177; 0.0006 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Petunidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6&lt;italic&gt;-p&lt;/italic&gt;-(coumaroyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0298 &amp;#177; 0.0022 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.029 &amp;#177; 0.0012 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0184 &amp;#177; 0.0089 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0068 &amp;#177; 0.0005 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0107 &amp;#177; 0.0012 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.006 &amp;#177; 0.0006 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Malvidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6-p-(coumaroyl)-glucoside (&lt;italic&gt;cis&lt;/italic&gt; isomer)&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0037 &amp;#177; 0.0004 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0041 &amp;#177; 0.0002 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0022 &amp;#177; 0.0013 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0016 &amp;#177; 0.0003 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0025 &amp;#177; 0.0003 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0013 &amp;#177; 0.0004 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Peonidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6-p-(coumaroyl)-glucoside&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0203 &amp;#177; 0.0019 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0226 &amp;#177; 0.0032 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0479 &amp;#177; 0.061 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0116 &amp;#177; 0.0015 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0181 &amp;#177; 0.0004 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0098 &amp;#177; 0.0011 &lt;sup&gt;c&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;Malvidin-3-&lt;italic&gt;O&lt;/italic&gt;-(6&lt;italic&gt;-p&lt;/italic&gt;-(coumaroyl)-glucoside (&lt;italic&gt;trans&lt;/italic&gt; isomer)&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.1753 &amp;#177; 0.0157 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.166 &amp;#177; 0.0069 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.097 &amp;#177; 0.057 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0455 &amp;#177; 0.0013 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0746 &amp;#177; 0.0047 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;0.0365 &amp;#177; 0.0062 &lt;sup&gt;c&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;&amp;#8721; c Anthocyanins (with unknown)&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;1.3222 &amp;#177; 0.1247 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;1.318 &amp;#177; 0.0299 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;1.0471 &amp;#177; 0.0597 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;0.4783 &amp;#177; 0.0241 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;0.7667 &amp;#177; 0.0806 &lt;sup&gt;a&lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;0.4572 &amp;#177; 0.0475 &lt;sup&gt;b&lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;/tbody&gt;&lt;/table&gt; </ephtml> Different superscript lower case letters in each row indicate statistically significant differences (<emph>p</emph> &lt; 0.001) between treatments.</p> <p>Table 4 Minimum inhibitory concentrations (MIC) of grape skin extracts against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus (mg mL−1).</p> <p> <ephtml> &lt;table&gt;&lt;thead&gt;&lt;tr&gt;&lt;th rowspan="2" align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;&lt;italic&gt;Vitis vinifera&lt;/italic&gt; L. variety&lt;/th&gt;&lt;th rowspan="2" align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Year&lt;/th&gt;&lt;th rowspan="2" align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Treatment&lt;/th&gt;&lt;th colspan="4" align="center" style="border-top:solid thin"&gt;MIC (mg mL&lt;sup&gt;&amp;#8722;1&lt;/sup&gt;)&lt;/th&gt;&lt;/tr&gt;&lt;tr&gt;&lt;th align="center" style="border-bottom:solid thin"&gt;&lt;italic&gt;E. coli&lt;/italic&gt;&lt;/th&gt;&lt;th align="center" style="border-bottom:solid thin"&gt;&lt;italic&gt;P. aeruginosa&lt;/italic&gt;&lt;/th&gt;&lt;th align="center" style="border-bottom:solid thin"&gt;&lt;italic&gt;B. subtilis&lt;/italic&gt;&lt;/th&gt;&lt;th align="center" style="border-bottom:solid thin"&gt;&lt;italic&gt;S. aureus&lt;/italic&gt;&lt;/th&gt;&lt;/tr&gt;&lt;/thead&gt;&lt;tbody&gt;&lt;tr&gt;&lt;td rowspan="6" align="center" valign="middle" style="border-bottom:solid thin"&gt;Blaufr&amp;#228;nkisch &lt;/td&gt;&lt;td rowspan="3" align="center" valign="middle"&gt;2015&lt;/td&gt;&lt;td align="center" valign="top"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.79 &amp;#177; 0.52&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;6.23 &amp;#177; 0.49&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.46 &amp;#177; 0.99&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.46 &amp;#177; 0.79&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="top"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;9.15 &amp;#177; 0.16&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;4.71 &amp;#177; 0.24&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;9.37 &amp;#177; 0.45&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;9.61 &amp;#177; 0.79&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="top"&gt;T3&lt;/td&gt;&lt;td align="center" valign="middle"&gt;9.96 &amp;#177; 0.34&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;5.05 &amp;#177; 0.11&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;10.06 &amp;#177; 0.24&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;9.96 &amp;#177; 0.34&lt;sub&gt; b &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td rowspan="3" align="center" valign="middle" style="border-bottom:solid thin"&gt;2016&lt;/td&gt;&lt;td align="center" valign="top"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.79 &amp;#177; 0.53&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.79 &amp;#177; 0.54 &lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.46 &amp;#177; 0.10&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;25.93 &amp;#177; 0.65&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="top"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;9.15 &amp;#177; 0.17&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;9.15 &amp;#177; 0.18&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;9.37 &amp;#177; 0.46&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;18.22 &amp;#177; 0.29&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; c &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="top" style="border-bottom:solid thin"&gt;T3&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;9.96 &amp;#177; 0.35&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;9.96 &amp;#177; 0.36&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;10.06 &amp;#177; 0.25&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;19.93 &amp;#177; 0.69&lt;sub&gt; a &lt;/sub&gt; A &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td rowspan="6" align="center" valign="middle" style="border-bottom:solid thin"&gt;Merlot &lt;/td&gt;&lt;td rowspan="3" align="center" valign="middle"&gt;2015&lt;/td&gt;&lt;td align="center" valign="top"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.72 &amp;#177; 0.66&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;4.03 &amp;#177; 0.36&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.73 &amp;#177; 0.67&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.61 &amp;#177; 0.58&lt;sub&gt; b &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="top"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;6.88 &amp;#177; 0.68&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;3.27 &amp;#177; 0.39&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;6.84 &amp;#177; 0.51&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.14 &amp;#177; 0.12&lt;sub&gt; b &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="top"&gt;T3&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.31 &amp;#177; 0.63&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a,b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;3.82 &amp;#177; 0.49&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.31 &amp;#177; 0.63&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a,b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.28 &amp;#177; 0.25&lt;sub&gt; b &lt;/sub&gt; B &lt;sup&gt; a,b &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td rowspan="3" align="center" valign="middle" style="border-bottom:solid thin"&gt;2016&lt;/td&gt;&lt;td align="center" valign="top"&gt;T1&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.74 &amp;#177; 0.67&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;4.13 &amp;#177; 0.37&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.83&amp;#177; 0.68&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;15.78 &amp;#177; 1.29&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="top"&gt;T2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;6.98 &amp;#177; 0.69&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;3.47 &amp;#177; 0.40&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;6.94 &amp;#177; 0.52&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle"&gt;13.75 &amp;#177; 1.26&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; b &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="top" style="border-bottom:solid thin"&gt;T3&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;7.41 &amp;#177; 0.64&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a,b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;3.92 &amp;#177; 0.50&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;7.41 &amp;#177; 0.64&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a,b &lt;/sup&gt;&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;15.18 &amp;#177; 0.37&lt;sub&gt; a &lt;/sub&gt; B &lt;sup&gt; a &lt;/sup&gt;&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td colspan="3" align="center" valign="middle" style="border-bottom:solid thin"&gt;Amikacine sulfate&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;0.000016&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;0.00003&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;0.000016&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;0.000008&lt;/td&gt;&lt;/tr&gt;&lt;/tbody&gt;&lt;/table&gt; </ephtml> Different green subscript lower case letters in each row indicate statistically significant differences (<emph>p</emph> &lt; 0.001) between years for the same variety and same treatment. Different blue upper case letters in each row indicate statistically significant differences (<emph>p</emph> &lt; 0.001) between varieties for the same year and same treatment. Different magenta superscript lower case letters in each row indicate statistically significant differences (<emph>p</emph> &lt; 0.001) between treatments for the same variety and same year.</p> <p>Table 5 Rainfall and mean temperatures in Đakovo from March to October in 2015 and 2016.</p> <p> <ephtml> &lt;table&gt;&lt;thead&gt;&lt;tr&gt;&lt;th rowspan="2" align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Month&lt;/th&gt;&lt;th colspan="2" align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Mean Monthly Temperature, &amp;#176;C&lt;/th&gt;&lt;th colspan="2" align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Mean Min-Max Temperature, &amp;#176;C&lt;/th&gt;&lt;th colspan="2" align="center" style="border-top:solid thin;border-bottom:solid thin"&gt;Rainfall, mm&lt;/th&gt;&lt;/tr&gt;&lt;tr&gt;&lt;th align="center" style="border-bottom:solid thin"&gt;2015&lt;/th&gt;&lt;th align="center" style="border-bottom:solid thin"&gt;2016&lt;/th&gt;&lt;th align="center" style="border-bottom:solid thin"&gt;2015&lt;/th&gt;&lt;th align="center" style="border-bottom:solid thin"&gt;2016&lt;/th&gt;&lt;th align="center" style="border-bottom:solid thin"&gt;2015&lt;/th&gt;&lt;th align="center" style="border-bottom:solid thin"&gt;2016&lt;/th&gt;&lt;/tr&gt;&lt;/thead&gt;&lt;tbody&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;March&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.8&lt;/td&gt;&lt;td align="center" valign="middle"&gt;7.9&lt;/td&gt;&lt;td align="center" valign="middle"&gt;2.7&amp;#8211;15.0&lt;/td&gt;&lt;td align="center" valign="middle"&gt;3.3&amp;#8211;17.6&lt;/td&gt;&lt;td align="center" valign="middle"&gt;48.5&lt;/td&gt;&lt;td align="center" valign="middle"&gt;82.7&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;April&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.7&lt;/td&gt;&lt;td align="center" valign="middle"&gt;13.7&lt;/td&gt;&lt;td align="center" valign="middle"&gt;5.9&amp;#8211;19.7&lt;/td&gt;&lt;td align="center" valign="middle"&gt;5.8&amp;#8211;19.2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;18.2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;46.5&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;May&lt;/td&gt;&lt;td align="center" valign="middle"&gt;18.2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;16.6&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.0&amp;#8211;25.4&lt;/td&gt;&lt;td align="center" valign="middle"&gt;9.4&amp;#8211;23.8&lt;/td&gt;&lt;td align="center" valign="middle"&gt;130.8&lt;/td&gt;&lt;td align="center" valign="middle"&gt;72.1&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;June&lt;/td&gt;&lt;td align="center" valign="middle"&gt;21.3&lt;/td&gt;&lt;td align="center" valign="middle"&gt;21.4&lt;/td&gt;&lt;td align="center" valign="middle"&gt;13.5&amp;#8211;27.0&lt;/td&gt;&lt;td align="center" valign="middle"&gt;17.1&amp;#8211;28.4&lt;/td&gt;&lt;td align="center" valign="middle"&gt;16.8&lt;/td&gt;&lt;td align="center" valign="middle"&gt;92.7&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;July&lt;/td&gt;&lt;td align="center" valign="middle"&gt;25.0&lt;/td&gt;&lt;td align="center" valign="middle"&gt;23.3&lt;/td&gt;&lt;td align="center" valign="middle"&gt;17.6&amp;#8211;30.0&lt;/td&gt;&lt;td align="center" valign="middle"&gt;14.0&amp;#8211;28.4&lt;/td&gt;&lt;td align="center" valign="middle"&gt;12.4&lt;/td&gt;&lt;td align="center" valign="middle"&gt;125.2&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;August&lt;/td&gt;&lt;td align="center" valign="middle"&gt;24.2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;21.0&lt;/td&gt;&lt;td align="center" valign="middle"&gt;17.6&amp;#8211;28.9&lt;/td&gt;&lt;td align="center" valign="middle"&gt;16.9&amp;#8211;26.8&lt;/td&gt;&lt;td align="center" valign="middle"&gt;64.9&lt;/td&gt;&lt;td align="center" valign="middle"&gt;51.0&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle"&gt;September&lt;/td&gt;&lt;td align="center" valign="middle"&gt;18.2&lt;/td&gt;&lt;td align="center" valign="middle"&gt;18.4&lt;/td&gt;&lt;td align="center" valign="middle"&gt;11.1&amp;#8211;28.0&lt;/td&gt;&lt;td align="center" valign="middle"&gt;11.8&amp;#8211;22.8&lt;/td&gt;&lt;td align="center" valign="middle"&gt;63.1&lt;/td&gt;&lt;td align="center" valign="middle"&gt;61.5&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;October&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;11.3&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;10.4&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;11.1&amp;#8211;17.7&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;5.6&amp;#8211;17.2&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;114.6&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;66.0&lt;/td&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;Mean temp., (&amp;#176;C)&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;17.33&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;16.58&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin" /&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin" /&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin" /&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin" /&gt;&lt;/tr&gt;&lt;tr&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;Cumulative rainfall, mm&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin" /&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin" /&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin" /&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin" /&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;469.3&lt;/td&gt;&lt;td align="center" valign="middle" style="border-bottom:solid thin"&gt;597.7&lt;/td&gt;&lt;/tr&gt;&lt;/tbody&gt;&lt;/table&gt; </ephtml> </p> <hd id="AN0137474727-23">Author Contributions</hd> <p>Conceptualization, V.P.; Methodology, V.P., M.D., T.K., U.B., F.S. and M.K.; Software, V.P., Validation, V.P.; Formal analysis, V.P., U.B., V.J.; Investigation, V.P., M.D. and T.K.; Resources, M.D., F.S.; Data curation, V.P.,U.B.; Writing—original draft preparation, V.P.; Writing—review and editing, V.P., F.S.; Visualization, V.P.; Supervision, V.P.; Project administration, M.D., V.J.; Funding acquisition, M.D.</p> <hd id="AN0137474727-24">Funding</hd> <p>This research received no external funding.</p> <hd id="AN0137474727-25">Conflicts of Interest</hd> <p>The authors declare no conflict of interest.</p> <ref id="AN0137474727-26"> <title> Footnotes </title> <blist> <bibl id="bib1" idref="ref1" type="bt">1</bibl> <bibtext> Sample Availability: Samples of the grape skin extracts are available from the authors, except for Blaufränkisch variety from 2015.</bibtext> </blist> </ref> <ref id="AN0137474727-27"> <title> References </title> <blist> <bibtext> Beckman C.H. Phenolic-storing cells: Keys to programmed cell death and periderm formation in wilt disease resistance and in general defence responses in plants?. Physiol. Mol. Plant Pathol. 2000; 57: 101-110. 10.1006/pmpp.2000.0287</bibtext> </blist> <blist> <bibl id="bib2" idref="ref2" type="bt">2</bibl> <bibtext> Graf B.A., Milbury P.E., Blumberg J.B. Flavonols, flavones, flavanones, and human health: Epidemiological evidence. J. Med. Food. 2005; 8: 281-290. 10.1089/jmf.2005.8.281. 16176136</bibtext> </blist> <blist> <bibl id="bib3" type="bt">3</bibl> <bibtext> Arts I.C.W., Hollman P.C.H. Polyphenols and disease risk in epidemiologic studies. Am. J. Clin. Nutr. 2005; 81: 317S-325S. 10.1093/ajcn/81.1.317S. 15640497</bibtext> </blist> <blist> <bibl id="bib4" idref="ref3" type="bt">4</bibl> <bibtext> Xia E.-Q., Deng G.-F., Guo Y.-J., Li H.-B. Biological activities of polyphenols from grapes. Int. J. Mol. Sci. 2010; 11: 622-646. 10.3390/ijms11020622. 20386657</bibtext> </blist> <blist> <bibl id="bib5" type="bt">5</bibl> <bibtext> Raj N.K., Sripal R.M., Chaluvadi M.R., Krishna D.R. Bioflavonoids classification, pharmacological, biochemical effects and therapeutic potential. Indian J. Pharmacol. 2001; 33: 2</bibtext> </blist> <blist> <bibl id="bib6" idref="ref39" type="bt">6</bibl> <bibtext> Heim K.E., Tagliaferro A.R., Bobilya D.J. Flavonoid antioxidants: Chemistry, metabolism and structure-activity relationships. J. Nutr. Biochem. 2002; 13: 572-584. 10.1016/S0955-2863(02)00208-5</bibtext> </blist> <blist> <bibl id="bib7" type="bt">7</bibl> <bibtext> Han X., Shen T., Lou H. Dietary Polyphenols and Their Biological Significance. Int. J. Mol. Sci. 2007; 8: 950-988. 10.3390/i8090950</bibtext> </blist> <blist> <bibl id="bib8" idref="ref4" type="bt">8</bibl> <bibtext> Seeram N.P., Cichewicz R.H., Chandra A., Nair M.G. Cyclooxygenase Inhibitory and Antioxidant Compounds from Crabapple Fruits. J. Agric. Food Chem. 2003; 51: 1948-1951. 10.1021/jf025993u. 12643656</bibtext> </blist> <blist> <bibl id="bib9" idref="ref5" type="bt">9</bibl> <bibtext> Cho M.J., Howard L.R., Prior R.L., Clark J.R. Flavonoid glycosides and antioxidant capacity of various blackberry, blueberry and red grape genotypes determined by high-performance liquid chromatography/mass spectrometry. J. Sci. Food Agric. 2004; 84: 1771-1782. 10.1002/jsfa.1885</bibtext> </blist> <blist> <bibtext> Letenneur L., Proust-Lima C., Le Gouge A., Dartigues J.F., Barberger-Gateau P. Flavonoid intake and cognitive decline over a 10-year period. Am. J. Epidemiol. 2007; 165: 1364-1371. 10.1093/aje/kwm036</bibtext> </blist> <blist> <bibtext> Scarmeas N., Luchsinger J.A., Mayeux R., Stern Y. Mediterranean diet and Alzheimer disease mortality. Neurology. 2007; 69: 1084-1093. 10.1212/01.wnl.0000277320.50685.7c. 17846408</bibtext> </blist> <blist> <bibtext> Ali K., Maltese F., Choi Y.H., Verpoorte R. Metabolic constituents of grapevine and grape-derived products. Phytochem. Rev. Proc. Phytochem. Soc. Eur. 2010; 9: 357-378. 10.1007/s11101-009-9158-0. 20835385</bibtext> </blist> <blist> <bibtext> Perumalla A.V.S., Hettiarachchy N.S. Green tea and grape seed extracts — Potential applications in food safety and quality. Food Res. Int. 2011; 44: 827-839. 10.1016/j.foodres.2011.01.022</bibtext> </blist> <blist> <bibtext> Jayaprakasha G.K., Selvi T., Sakariah K.K. Antibacterial and antioxidant activities of grape (Vitis vinifera) seed extracts. Food Res. Int. 2002; 36: 117-122. 10.1016/S0963-9969(02)00116-3</bibtext> </blist> <blist> <bibtext> Rhodes P.L., Mitchell J.W., Wilson M.W., Melton L.D. Antilisterial activity of grape juice and grape extracts derived from Vitis vinifera variety Ribier. Int. J. Food Microbiol. 2006; 107: 281-286. 10.1016/j.ijfoodmicro.2005.10.022. 16386816</bibtext> </blist> <blist> <bibtext> Brown J.C., Huang G., Haley-Zitlin V., Jiang X. Antibacterial Effects of Grape Extracts on Helicobacter pylori. Appl. Environ. Microbiol. 2009; 75: 848-852. 10.1128/AEM.01595-08</bibtext> </blist> <blist> <bibtext> Sternad Lemut M., Trost K., Sivilotti P., Vrhovsek U. Pinot Noir grape colour related phenolics as affected by leaf removal treatments in the Vipava Valley. J. Food Compos. Anal. 2011; 24: 777-784. 10.1016/j.jfca.2011.03.003</bibtext> </blist> <blist> <bibtext> Reynolds A.G., Wardle D.A., Hall J.W., Dever M. Fruit Maturation of Four Vitis vinifera Cultivars in Response to Vineyard Location and Basal Leaf Removal. Am. J. Enol. Vitic. 1995; 46: 542-558</bibtext> </blist> <blist> <bibtext> Matus J.T., Poupin M.J., Cañón P., Bordeu E., Alcalde J.A., Arce-Johnson P. Isolation of WDR and bHLH genes related to flavonoid synthesis in grapevine (Vitis vinifera L.). Plant Mol. Biol. 2010; 72: 607-620. 10.1007/s11103-010-9597-4</bibtext> </blist> <blist> <bibtext> Pereira G.E., Gaudillere J.-P., Pieri P., Hilbert G., Maucourt M., Deborde C., Moing A., Rolin D. Microclimate influence on mineral and metabolic profiles of grape berries. J. Agric. Food Chem. 2006; 54: 6765-6775. 10.1021/jf061013k</bibtext> </blist> <blist> <bibtext> Guidoni S., Ferrandino A., Novello V. Effects of Seasonal and Agronomical Practices on Skin Anthocyanin Profile of Nebbiolo Grapes. Am. J. Enol. Vitic. 2008; 59: 22-29</bibtext> </blist> <blist> <bibtext> Teixeira A., Eiras-Dias J., Castellarin S.D., Gerós H. Berry phenolics of grapevine under challenging environments. Int. J. Mol. Sci. 2013; 14: 18711-18739. 10.3390/ijms140918711. 24030720</bibtext> </blist> <blist> <bibtext> Verzera A., Tripodi G., Dima G., Condurso C., Scacco A., Cincotta F., Giglio D.M.L., Santangelo T., Sparacio A. Leaf removal and wine composition of Vitis vinifera L. cv. Nero d'Avola: The volatile aroma constituents. J. Sci. Food Agric. 2016; 96: 150-159. 10.1002/jsfa.7075. 25581439</bibtext> </blist> <blist> <bibtext> Buesa I., Caccavello G., Basile B., Merli M.C., Poni S., Chirivella C., Intrigliolo D.S. Delaying berry ripening of Bobal and Tempranillo grapevines by late leaf removal in a semi-arid and temperate-warm climate under different water regimes. Aust. J. Grape Wine Res. 2019; 25: 70-82. 10.1111/ajgw.12368</bibtext> </blist> <blist> <bibtext> Lanari V., Lattanzi T., Borghesi L., Silvestroni O., Palliotti A. Post-Veraison Mechanical Leaf Removal Delays Berry Ripening on 'Sangiovese' and 'Montepulciano' Grapevines. Proceedings of the Proceedings of the 1st International Workshop on Vineyard Mechanization and Grape and Wine Quality. Piacenza, Italy. 27–29 June 2012: 327-333</bibtext> </blist> <blist> <bibtext> Zhang Z.-Z., Che X.-N., Pan Q.-H., Li X.-X., Duan C.-Q. Transcriptional activation of flavan-3-ols biosynthesis in grape berries by UV irradiation depending on developmental stage. Plant Sci. Int. J. Exp. Plant Biol. 2013; 208: 64-74. 10.1016/j.plantsci.2013.03.013</bibtext> </blist> <blist> <bibtext> Kennedy J.A., Matthews M.A., Waterhouse A.L. Effect of Maturity and Vine Water Status on Grape Skin and Wine Flavonoids. Am. J. Enol. Vitic. 2002; 53: 268-274</bibtext> </blist> <blist> <bibtext> Zhu L., Zhang Y., Lu J. Phenolic contents and compositions in skins of red wine grape cultivars among various genetic backgrounds and originations. Int. J. Mol. Sci. 2012; 13: 3492-3510. 10.3390/ijms13033492</bibtext> </blist> <blist> <bibtext> Yin H.-T., Tian Q.-Z., Guan L., Zhou Y., Huang X.-E., Zhang H. In vitro and in vivo evaluation of the antitumor efficiency of resveratrol against lung cancer. Asian Pac. J. Cancer Prev. APJCP. 2013; 14: 1703-1706. 10.7314/APJCP.2013.14.3.1703</bibtext> </blist> <blist> <bibtext> Fan G.-J., Liu X.-D., Qian Y.-P., Shang Y.-J., Li X.-Z., Dai F., Fang J.-G., Jin X.-L., Zhou B. 4,4′-Dihydroxy-trans-stilbene, a resveratrol analogue, exhibited enhanced antioxidant activity and cytotoxicity. Bioorg. Med. Chem. 2009; 17: 2360-2365. 10.1016/j.bmc.2009.02.014</bibtext> </blist> <blist> <bibtext> Bonnefont-Rousselot D. Resveratrol and Cardiovascular Diseases. Nutrients. 2016; 8. 10.3390/nu8050250</bibtext> </blist> <blist> <bibtext> Drenjančević M., Jukić V., Zmaić K., Kujundžić T., Rastija V. Effects of early leaf removal on grape yield, chemical characteristics, and antioxidant activity of grape variety Cabernet Sauvignon and wine from eastern Croatia. Acta Agric. Scand. Sect. B Soil Plant Sci. 2017; 67: 705-711</bibtext> </blist> <blist> <bibtext> Poni S., Casalini L., Bernizzoni F., Civardi S., Intrieri C. Effects of Early Defoliation on Shoot Photosynthesis, Yield Components, and Grape Composition. Am. J. Enol. Vitic. 2006; 57: 397-407</bibtext> </blist> <blist> <bibtext> Risco D., Pérez D., Yeves A., Castel J.R., Intrigliolo D.S. Early defoliation in a temperate warm and semi-arid Tempranillo vineyard: Vine performance and grape composition. Aust. J. Grape Wine Res. 2014; 20: 111-122. 10.1111/ajgw.12049</bibtext> </blist> <blist> <bibtext> Mijowska K., Ochmian I., Oszmiański J. Impact of Cluster Zone Leaf Removal on Grapes cv. Regent Polyphenol Content by the UPLC-PDA/MS Method. Molecules. 2016; 21. 10.3390/molecules21121688. 27973426</bibtext> </blist> <blist> <bibtext> Jerman T., Lemut M.S., Trôst K., Pospišil M. The impact of early leaf removal on polyphenol/anthocyanin content and in vitro antioxidant potential of "Pinot Noir" grapes from Vipava Valley. Proceedings of 46th Croatian and 6th International Symposium on Agriculture. Opatija, Croatia. 14–18 February 2011: 940</bibtext> </blist> <blist> <bibtext> Jones G.V., Davis R.E. Climate Influences on Grapevine Phenology, Grape Composition, and Wine Production and Quality for Bordeaux, France. Am. J. Enol. Vitic. 2000; 51: 249-261</bibtext> </blist> <blist> <bibtext> Ferrer M., Echeverría G., Carbonneau A. Effect of berry weight and its components on the contents of sugars and anthocyanins of three varieties of Vitis vinifera L. under different water supply conditions. South Afr. J. Enol. Vitic. 2014; 35: 103-113. 10.21548/35-1-989</bibtext> </blist> <blist> <bibtext> Barbagallo M.G., Guidoni S., Hunter J.J. Berry Size and Qualitative Characteristics of Vitis vinifera L. cv. Syrah. South Afr. J. Enol. Vitic. 2011; 32: 129-136. 10.21548/32-1-1372</bibtext> </blist> <blist> <bibtext> Roby G., Harbertson J.F., Adams D.A., Matthews M.A. Berry size and vine water deficits as factors in winegrape composition: Anthocyanins and tannins. Aust. J. Grape Wine Res. 2004; 10: 100-107. 10.1111/j.1755-0238.2004.tb00012.x</bibtext> </blist> <blist> <bibtext> Nirmala J.G., Narendhirakannan R.T. In Vitro Antioxidant And Antimicrobial Activities Of Grapes (Vitis Vinifera. L) Seed And Skin Extracts - Muscat Variety. Int. J. Pharm. Pharm. Sci. 2011; 3: 8</bibtext> </blist> <blist> <bibtext> Intrigliolo D.S., Llacer E., Revert J., Esteve M.D., Climent M.D., Palau D., Gómez I. Early defoliation reduces cluster compactness and improves grape composition in Mandó, an autochthonous cultivar of Vitis vinifera from southeastern Spain. Sci. Hortic. 2014; 167: 71-75. 10.1016/j.scienta.2013.12.036</bibtext> </blist> <blist> <bibtext> Ribureau-Gayon P. The anthocyanins of the genus Vitis. Application to the differentiation of wines. C. r. Hebd. Seances Acad. Sci. 1960; 250: 591-593</bibtext> </blist> <blist> <bibtext> Mira de Orduña R. Climate change associated effects on grape and wine quality and production. Food Res. Int. 2010; 43: 1844-1855. 10.1016/j.foodres.2010.05.001</bibtext> </blist> <blist> <bibtext> Castellarin S.D., Bavaresco L., Falginella L., Goncalves M.I.V.Z., Di Gaspero G.. Phenolics in Grape Berry and Key Antioxidants; Bentham Science Publishers: Sharjah, United Arab Emirates. 2012. 978-1-60805-360-5</bibtext> </blist> <blist> <bibtext> Katalinić V., Možina S.S., Skroza D., Generalić I., Abramovič H., Miloš M., Ljubenkov I., Piskernik S., Pezo I., Terpinc P. Polyphenolic profile, antioxidant properties and antimicrobial activity of grape skin extracts of 14 Vitis vinifera varieties grown in Dalmatia (Croatia). Food Chem. 2010; 119: 715-723. 10.1016/j.foodchem.2009.07.019</bibtext> </blist> <blist> <bibtext> Lorenz D.H., Eichhorn K.W., Bleiholder H., Klose R., Meier U., Weber E. Growth Stages of the Grapevine: Phenological growth stages of the grapevine (Vitis vinifera L. ssp. vinifera)—Codes and descriptions according to the extended BBCH scale†. Aust. J. Grape Wine Res. 1995; 1: 100-103. 10.1111/j.1755-0238.1995.tb00085.x</bibtext> </blist> <blist> <bibtext> Rustioni L., Maghradze D., Popescu C.F., Cola G., Abashidze E., Aroutiounian R., Brazao J., Coletti S., Cornea V., Dejeu L. First results of the European grapevine collections' collaborative network: Validation of a standard eno-carpological phenotyping method. Vitis. 2014; 53: 219-226</bibtext> </blist> <blist> <bibtext> Singleton V.L., Rossi J.A. Colorimetry of Total Phenolics with Phosphomolybdic-Phosphotungstic Acid Reagents. Am. J. Enol. Vitic. 1965; 16: 144-158</bibtext> </blist> <blist> <bibtext> Nagel C.W., Wulf L.W. Changes in the Anthocyanins, Flavonoids and Hydroxycinnamic Acid Esters during Fermentation and Aging of Merlot and Cabernet Sauvignon. Am. J. Enol. Vitic. 1979; 30: 111-116</bibtext> </blist> <blist> <bibtext> Shih M.-H., Su Y.-S., Wu C.-L. Syntheses of Aromatic Substituted Hydrazino-thiazole Derivatives to Clarify Structural Characterization and Antioxidant Activity between 3-Arylsydnonyl and Aryl Substituted Hydrazino-thiazoles. Chem. Pharm. Bull. (Tokyo). 2007; 55: 1126-1135. 10.1248/cpb.55.1126</bibtext> </blist> <blist> <bibtext> OIV. Compendium of International Methods of Analysis of Wines and Musts (2 vol.); OIV: Paris, France. 2017</bibtext> </blist> <blist> <bibtext> Burns J., Mullen W., Landrault N., Teissedre P.-L., Lean M.E.J., Crozier A. Variations in the Profile and Content of Anthocyanins in Wines Made from Cabernet Sauvignon and Hybrid Grapes. J. Agric. Food Chem. 2002; 50: 4096-4102. 10.1021/jf011233s</bibtext> </blist> <blist> <bibtext> Ryan J.-M., Revilla E. Anthocyanin composition of Cabernet Sauvignon and Tempranillo grapes at different stages of ripening. J. Agric. Food Chem. 2003; 51: 3372-3378. 10.1021/jf020849u</bibtext> </blist> <blist> <bibtext> Radovanović B., Radovanović A. Free radical scavenging activity and anthocyanin profile of Cabernet Sauvignon wines from the Balkan region. Molecules. 2010; 15: 4213-4226. 10.3390/molecules15064213. 20657436</bibtext> </blist> <blist> <bibtext> Li Y., Ma R., Xu Z., Wang J., Chen T., Chen F., Wang Z. Identification and quantification of anthocyanins in Kyoho grape juice-making pomace, Cabernet Sauvignon grape winemaking pomace and their fresh skin. J. Sci. Food Agric. 2013; 93: 1404-1411. 10.1002/jsfa.5907. 23400926</bibtext> </blist> <blist> <bibtext> Gu W., Wang S. Synthesis and antimicrobial activities of novel 1H-dibenzo[a,c]carbazoles from dehydroabietic acid. Eur. J. Med. Chem. 2010; 45: 4692-4696. 10.1016/j.ejmech.2010.07.038. 20702006</bibtext> </blist> <blist> <bibtext> Pavić V., Flačer D., Jakovljević M., Molnar M., Jokić S. Assessment of Total Phenolic Content, In Vitro Antioxidant and Antibacterial Activity of Ruta graveolens L. Extracts Obtained by Choline Chloride Based Natural Deep Eutectic Solvents. Plants. 2019; 8. 10.3390/plants8030069. 30889869</bibtext> </blist> </ref> <aug> <p>By Valentina Pavić; Toni Kujundžić; Marina Kopić; Vladimir Jukić; Ulrike Braun; Florian Schwander; Mato Drenjančević; Margarida Castell Escuer and Mariona Camps-Bossacoma</p> <p>Reported by Author; Author; Author; Author; Author; Author; Author; Author; Author</p> </aug> <nolink nlid="nl1" bibid="bib10" firstref="ref6"></nolink> <nolink nlid="nl2" bibid="bib11" firstref="ref7"></nolink> <nolink nlid="nl3" bibid="bib12" firstref="ref8"></nolink> <nolink nlid="nl4" bibid="bib13" firstref="ref10"></nolink> <nolink nlid="nl5" bibid="bib14" firstref="ref11"></nolink> <nolink nlid="nl6" bibid="bib15" firstref="ref12"></nolink> <nolink nlid="nl7" bibid="bib16" firstref="ref13"></nolink> <nolink nlid="nl8" bibid="bib17" firstref="ref14"></nolink> <nolink nlid="nl9" bibid="bib18" firstref="ref15"></nolink> <nolink nlid="nl10" bibid="bib19" firstref="ref16"></nolink> <nolink nlid="nl11" bibid="bib22" firstref="ref18"></nolink> <nolink nlid="nl12" bibid="bib23" firstref="ref19"></nolink> <nolink nlid="nl13" bibid="bib24" firstref="ref20"></nolink> <nolink nlid="nl14" bibid="bib25" firstref="ref21"></nolink> <nolink nlid="nl15" bibid="bib26" firstref="ref22"></nolink> <nolink nlid="nl16" bibid="bib27" firstref="ref23"></nolink> <nolink nlid="nl17" bibid="bib28" firstref="ref24"></nolink> <nolink nlid="nl18" bibid="bib29" firstref="ref25"></nolink> <nolink nlid="nl19" bibid="bib32" firstref="ref26"></nolink> <nolink nlid="nl20" bibid="bib33" firstref="ref28"></nolink> <nolink nlid="nl21" bibid="bib34" firstref="ref29"></nolink> <nolink nlid="nl22" bibid="bib35" firstref="ref31"></nolink> <nolink nlid="nl23" bibid="bib36" firstref="ref32"></nolink> <nolink nlid="nl24" bibid="bib37" firstref="ref34"></nolink> <nolink nlid="nl25" bibid="bib38" firstref="ref35"></nolink> <nolink nlid="nl26" bibid="bib39" firstref="ref36"></nolink> <nolink nlid="nl27" bibid="bib40" firstref="ref38"></nolink> <nolink nlid="nl28" bibid="bib41" firstref="ref42"></nolink> <nolink nlid="nl29" bibid="bib42" firstref="ref43"></nolink> <nolink nlid="nl30" bibid="bib43" firstref="ref44"></nolink> <nolink nlid="nl31" bibid="bib44" firstref="ref45"></nolink> <nolink nlid="nl32" bibid="bib46" firstref="ref47"></nolink> <nolink nlid="nl33" bibid="bib47" firstref="ref48"></nolink> <nolink nlid="nl34" bibid="bib48" firstref="ref49"></nolink> <nolink nlid="nl35" bibid="bib49" firstref="ref50"></nolink> <nolink nlid="nl36" bibid="bib50" firstref="ref51"></nolink> <nolink nlid="nl37" bibid="bib51" firstref="ref52"></nolink> <nolink nlid="nl38" bibid="bib52" firstref="ref55"></nolink> <nolink nlid="nl39" bibid="bib53" firstref="ref56"></nolink> <nolink nlid="nl40" bibid="bib57" firstref="ref57"></nolink> <nolink nlid="nl41" bibid="bib58" firstref="ref58"></nolink> CustomLinks: – Url: https://resolver.ebsco.com/c/xy5jbn/result?sid=EBSCO:edsdoj&genre=article&issn=14203049&ISBN=&volume=24&issue=13&date=20190701&spage=2444&pages=2444-2444&title=Molecules&atitle=Effects%20of%20Defoliation%20on%20Phenolic%20Concentrations%2C%20Antioxidant%20and%20Antibacterial%20Activity%20of%20Grape%20Skin%20Extracts%20of%20the%20Varieties%20Blaufr%C3%A4nkisch%20and%20Merlot%20%28Vitis%20vinifera%20L.%29&aulast=Valentina%20Pavi%C4%87&id=DOI:10.3390/molecules24132444 Name: Full Text Finder (for New FTF UI) (s8985755) Category: fullText Text: Find It @ SCU Libraries MouseOverText: Find It @ SCU Libraries – Url: https://doaj.org/article/f38842e967dd4472b0fb67151263d5bd Name: EDS - DOAJ (s8985755) Category: fullText Text: View record from DOAJ MouseOverText: View record from DOAJ
Header	DbId: edsdoj DbLabel: Directory of Open Access Journals An: edsdoj.f38842e967dd4472b0fb67151263d5bd RelevancyScore: 879 AccessLevel: 3 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 878.767700195313
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Items	– Name: Title Label: Title Group: Ti Data: Effects of Defoliation on Phenolic Concentrations, Antioxidant and Antibacterial Activity of Grape Skin Extracts of the Varieties Blaufränkisch and Merlot (Vitis vinifera L.) – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Valentina+Pavić%22">Valentina Pavić</searchLink><br /><searchLink fieldCode="AR" term="%22Toni+Kujundžić%22">Toni Kujundžić</searchLink><br /><searchLink fieldCode="AR" term="%22Marina+Kopić%22">Marina Kopić</searchLink><br /><searchLink fieldCode="AR" term="%22Vladimir+Jukić%22">Vladimir Jukić</searchLink><br /><searchLink fieldCode="AR" term="%22Ulrike+Braun%22">Ulrike Braun</searchLink><br /><searchLink fieldCode="AR" term="%22Florian+Schwander%22">Florian Schwander</searchLink><br /><searchLink fieldCode="AR" term="%22Mato+Drenjančević%22">Mato Drenjančević</searchLink> – Name: TitleSource Label: Source Group: Src Data: Molecules, Vol 24, Iss 13, p 2444 (2019) – Name: Publisher Label: Publisher Information Group: PubInfo Data: MDPI AG, 2019. – Name: DatePubCY Label: Publication Year Group: Date Data: 2019 – Name: Subset Label: Collection Group: HoldingsInfo Data: LCC:Organic chemistry – Name: Subject Label: Subject Terms Group: Su Data: <searchLink fieldCode="DE" term="%22defoliation%22">defoliation</searchLink><br /><searchLink fieldCode="DE" term="%22Blaufränkisch%22">Blaufränkisch</searchLink><br /><searchLink fieldCode="DE" term="%22Merlot%22">Merlot</searchLink><br /><searchLink fieldCode="DE" term="%22phenols%22">phenols</searchLink><br /><searchLink fieldCode="DE" term="%22antioxidant+activity%22">antioxidant activity</searchLink><br /><searchLink fieldCode="DE" term="%22antibacterial+activity%22">antibacterial activity</searchLink><br /><searchLink fieldCode="DE" term="%22Organic+chemistry%22">Organic chemistry</searchLink><br /><searchLink fieldCode="DE" term="%22QD241-441%22">QD241-441</searchLink> – Name: Abstract Label: Description Group: Ab Data: Vitis vinifera L. has been highlighted by its many positive effects on human health, including antibacterial, anti-inflammatory and antioxidant activity due to its rich phytochemical content. Removing six basal leaves has great potential to influence the production of phytochemicals. The purpose of this study was to determine the impact of different terms of defoliation on the antioxidant and antibacterial activity of grape skin extracts of the Blaufränkisch and Merlot varieties. The total phenolic concentration, total and individual anthocyanin concentration, antioxidant activity and antibacterial activity on gram-positive and gram-negative human pathogens have been determined. The study was conducted on three treatments (control treatment without defoliation, defoliation immediately after bloom and defoliation before the start of the véraison phase). The results showed statistically significant enhancement of the total phenolic concentration as well as the antioxidant and antibacterial activity in both studied cultivars. Defoliation just after blooming was the preferable defoliation term in the Merlot variety for achieving the highest total anthocyanin concentration, antioxidant activity and significant increase of antibacterial activity against all four investigated bacteria. Defoliation before the start of the véraison phase was the preferable defoliation term for achieving the highest total anthocyanin concentration in the Blaufränkisch variety. In general, treatment of defoliation immediately after bloom was more beneficial compared with the defoliation before the start of the véraison phase. – Name: TypeDocument Label: Document Type Group: TypDoc Data: article – Name: Format Label: File Description Group: SrcInfo Data: electronic resource – Name: Language Label: Language Group: Lang Data: English – Name: ISSN Label: ISSN Group: ISSN Data: 1420-3049 – Name: NoteTitleSource Label: Relation Group: SrcInfo Data: https://www.mdpi.com/1420-3049/24/13/2444; https://doaj.org/toc/1420-3049 – Name: DOI Label: DOI Group: ID Data: 10.3390/molecules24132444 – Name: URL Label: Access URL Group: URL Data: <link linkTarget="URL" linkTerm="https://doaj.org/article/f38842e967dd4472b0fb67151263d5bd" linkWindow="_blank">https://doaj.org/article/f38842e967dd4472b0fb67151263d5bd</link> – Name: AN Label: Accession Number Group: ID Data: edsdoj.f38842e967dd4472b0fb67151263d5bd
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RecordInfo	BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.3390/molecules24132444 Languages: – Text: English PhysicalDescription: Pagination: PageCount: 1 StartPage: 2444 Subjects: – SubjectFull: defoliation Type: general – SubjectFull: Blaufränkisch Type: general – SubjectFull: Merlot Type: general – SubjectFull: phenols Type: general – SubjectFull: antioxidant activity Type: general – SubjectFull: antibacterial activity Type: general – SubjectFull: Organic chemistry Type: general – SubjectFull: QD241-441 Type: general Titles: – TitleFull: Effects of Defoliation on Phenolic Concentrations, Antioxidant and Antibacterial Activity of Grape Skin Extracts of the Varieties Blaufränkisch and Merlot (Vitis vinifera L.) Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Valentina Pavić – PersonEntity: Name: NameFull: Toni Kujundžić – PersonEntity: Name: NameFull: Marina Kopić – PersonEntity: Name: NameFull: Vladimir Jukić – PersonEntity: Name: NameFull: Ulrike Braun – PersonEntity: Name: NameFull: Florian Schwander – PersonEntity: Name: NameFull: Mato Drenjančević IsPartOfRelationships: – BibEntity: Dates: – D: 01 M: 07 Type: published Y: 2019 Identifiers: – Type: issn-print Value: 14203049 Numbering: – Type: volume Value: 24 – Type: issue Value: 13 Titles: – TitleFull: Molecules Type: main
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