## 3 Results and Discussion
### Bread Quality Characteristics  
Due to the low fungistatic activity obtained with the 5% addition of _Calendula officinalis_ water extract in the culture medium, it was decided to use only the higher addition in bread production and replace the water recommended by the recipe with 10% and 15% extract. The evaluation of the control and bread with the addition of extract quality parameters is shown in Table 2. The appearance of the prepared bread is shown in Figure 3.  
The addition of _Calendula officinalis_ water extract used in the study did not affect bread yields, and the results were similar to the control. The experiments found that bread with the extract showed significantly lower baking losses compared to the control. The total baking loss depends on numerous factors, including the size, shape, volume, and type of bread, dough management and baking method, and even the type of oven used [42]. [PERSON] et al. [43] report that the lower baking losses in bread with the addition of marigold may be due to the presence of a large amount of lutein, which binds to water and thus reduces moisture loss. Accordingly, it was found that the enriched bread had a significantly higher (about 18%) moisture content compared to the control. The bread with extract also had significantly higher volume than the control, by about 23% and 26% in the bread, in which 15% and 10% of the water was replaced with _Calendula officinalis_ extract, respectively. The specific bread volume is considered one of the main characteristics defining consumer acceptability [43], so the results obtained in this paper are significant. In contrast to the present study, [PERSON] et al. [43] found that enriching bread with dried marigold flower  
\begin{table}
\begin{tabular}{l c c c} \hline \hline \multirow{2}{*}{**Parameters**} & \multicolumn{3}{c}{**Sample**} \\ \cline{2-4}  & **Control** & **N10** & **N15** \\ \hline Bread yield (\%) & 139.54 \(\pm\) 2.78 a & 139.42 \(\pm\) 2.60 a & 140.49 \(\pm\) 0.73 a \\ \hline Baking loss (\%) & 11.14 \(\pm\) 1.11 a & 10.68 \(\pm\) 0.39 b & 9.16 \(\pm\) 1.46 c \\ \hline Specific volume (cm\({}^{3}\)/g) & 2.12 \(\pm\) 0.19 b & 2.67 \(\pm\) 0.17 a & 2.61 \(\pm\) 0.23 a \\ \hline Crumb moisture (\%) & 31.64 \(\pm\) 3.29 b & 37.26 \(\pm\) 5.40 a & 37.49 \(\pm\) 5.89 a \\ \hline Acidity [\({}^{\circ}\)] & 1.73 \(\pm\) 0.12 c & 2.00 \(\pm\) 0.00 b & 2.23 \(\pm\) 0.03 a \\ \hline \hline \end{tabular} N10, N15—bread, in which 10% and 15% of the water have been replaced with _Calendula officinalis_ water extract, respectively. Within each row, values with the same letter do not differ significantly at \(p<0.05\) ([PERSON]’s post hoc test); mean value (\(n=3\)) \(\pm\) SD.  
\end{table}
Table 2: Evaluation of bread quality parameters.  
powder reduced volume. In the experiments presented here, it was also noted that the acidity of the bread increased as the addition of extract increased.  
Dough expansion depends on dough rheological characteristics, primarily due to the gluten and starch components. Adding bioactive ingredients may physically modify the development of bread dough [44] since antioxidants can form complexes with proteins and/or polysaccharides. Several studies have shown an adverse effect of the powdered plant material addition on bread volume [45]. This can be explained by the presence of fibre, which interacts with proteins and weakens the gluten network of the dough, resulting in a reduction in the amount of CO\({}_{2}\) retained in the dough during fermentation [46]. [PERSON] et al. [47] report that _Calendula officinalis_ has a high total fibre content (62.33 \(\pm\) 9.17 g/100 g), but most of the fraction is insoluble (57.54 \(\pm\) 8.32 g/100 g). In general, insoluble fibres reduce the technological quality of baked goods, while soluble fibres have a positive effect [48], so using water extract was reasonable in the present study.