# The Impact of Grafted Larvae and Collection Day on Royal Jelly's Production and Quality
## 3 Results and Discussion
### Effect of Larval Sex on Royal Jelly Production and Composition  
A phenomenon occasionally observed in honeybee colonies is the construction of queen cells that contain drone larvae [36]. This typically occurs in queenless colonies  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**Parameter** & **Group A2** & **Group B2** & **Group C2** \\  & **(_n_ = 12)** & **(_n_ = 12)** & **(_n_ = 12)** \\ \hline Yield per Royal Cell & 0.193 \(\pm\) 0.06  & 0.241 \(\pm\) 0.09  & 68.2 \(\pm\) 2.00  \\ (\%) & (60.5–69.7) & (64.8–71.8) & (64.9–69.8) \\ \hline Protein Content & 12.4 \(\pm\) 0.72  & 12.5 \(\pm\) 0.43  & 3.62 \(\pm\) 0.63  \\ (\%) & (2.85–4.89) & (2.80–4.85) & (2.81–4.90) \\ \hline Glucose & 3.88 \(\pm\) 0.81  & 3.19 \(\pm\) 0.32  & 3.12 \(\pm\) 0.86  \\ (\%) & (2.09–4.00) & (1.99–4.28) & (1.76–3.64) \\ \hline
10-HDA & 3.21 \(\pm\) 0.40  & 3.41 \(\pm\) 0.27 [FOOTNOTE:]Footnote : Where different latin letters are used, statistically significant differences were found (a = 0.05). The values in parentheses refer to the range of values for each parameter. Where _n_: the number of samples in each group. A2 Group: 24 h larva; B2 Group: 48 h larva; C2 Group: 72 h larva. [ENDFOOTNOTE] \\ (\%) & (2.33–3.85) & (3.01–3 that have remained without a queen for an extended period and have developed laying workers. Additionally, during grafting, beekeepers are often unable to distinguish whether the transferred larvae are male (drone) or female (worker). The present experiment aimed to evaluate whether the RJ produced in queen cells grafted with drone larvae differs in quantity or quality from that produced in cells grafted with worker larvae. Specifically, the experiment tested whether worker bees perceive and respond differently to the sex of the grafted larvae in terms of RJ provisioning. The mean yield of RJ from artificial queen cells grafted with worker larvae was comparable to that from cells grafted with drone larvae (Table 3). Across all grafting trials, the observed RJ yield ranged from a minimum of 1.2 g to a maximum of 6.8 g per pooled sample. Notably, cells grafted with worker larvae exhibited a low coefficient of variation, indicating consistent RJ production, whereas those grafted with drone larvae showed a high coefficient of variation, suggesting substantial variability in yield. Although the total amount of RJ produced from worker-derived cells was higher than that from drone-derived cells, statistical analysis revealed no significant differences. Overall, the results indicate that the sex of the grafted larvae does not significantly influence RJ yield. Worker bees exhibited a similar acceptance rate for queen cells grafted with either worker or drone larvae, with both groups showing an overall acceptance rate of 64%. Across all grafting sessions, acceptance varied considerably, ranging from 30% (9 out of 30 grafted cells) to 83%, regardless of larval sex. Both types of grafted larvae displayed high coefficients of variation in acceptance, indicating that acceptance is not consistent and can fluctuate significantly, independent of the sex of the grafted larva. These findings suggest that even if a beekeeper inadvertently grafts a drone larva, RJ yield will not be significantly affected. The results align with previous studies by [PERSON] et al. (2017), which also concluded that larval sex does not influence acceptance or RJ production. The average moisture content of RJ did not differ significantly between cells grafted with worker or drone larvae. The recorded moisture content ranged from 67.8% to 71.8%. Similarly, the average protein content remained comparable between both groups, with