# Comparative Analysis of Urban and Rural Ecosystems in the Context of Suitability for Honeybees
## 2 Materials and Methods  
### Experimental Setup  
The study was conducted in five regions of Poland in 2021 and 2022 (see Table 1 and Figure 1). In each region, we located an experimental apiary in the largest city or most urbanised area (Urban) and control apiaries in agricultural areas at varying distances from experimental apiaries, depending on the region, between 4.5 and 39.5 km (Rural). Tychy was only included in 2021, and Cracow only in 2022. The regions are characterised by a relatively low elevation, with altitudes ranging from 101 to 277 m above sea level. The mean annual precipitation in Poland is approximately 608.8 mm. Poland is located within a temperate climate zone, with an average summer temperature of approximately 18 \({}^{\circ}\)C and an average winter temperature in winter \(-\)1.5 \({}^{\circ}\)C. The selected sites have been identified as those which exhibit minimal variation in terms of climate.  
\begin{table}
\begin{tabular}{p{56.9 pt} p{56.9 pt} p{56.9 pt} p{56.9 pt} p{56.9 pt} p{56.9 pt}} \hline \hline \multirow{2}{*}{**No. of Apiary**} & \multirow{2}{*}{**Location of the Urban Apiary (City)**} & \multirow{2}{*}{**Coordinates of Urban Apiary**} & \multicolumn{2}{p{56.9 pt}}{**Distance (km) and Direction to the Control Apiary (Rural)**} & \multicolumn{2}{p{56.9 pt}}{**The Mean Annual**} \\  & & & **Direction to the Rural** & **Annual Temperature (\({}^{\circ}\)C)** & **Rainfall (mm)** \\ \hline \multirow{2}{*}{1} & \multirow{2}{*}{Warsaw} & 52\({}^{\circ}\)17\({}^{\prime}\)05.0\({}^{\prime\prime}\) N & \multirow{2}{*}{39.5 km, S} & \multirow{2}{*}{8.5} & \multirow{2}{*}{530} \\  & & 20\({}^{\circ}\)55\({}^{\prime}\)52.0\({}^{\prime\prime}\) E & & & \\  & & 51\({}^{\circ}\)14\({}^{\prime}\)48.6\({}^{\prime\prime}\) N & \multirow{2}{*}{22.4 km W} & \multirow{2}{*}{8.3} & \multirow{2}{*}{560} \\  & & 22\({}^{\circ}\)32\({}^{\prime}\)54.7\({}^{\prime\prime}\) E & & & \\  & & 51\({}^{\circ}\)24\({}^{\prime}\)35.7\({}^{\prime\prime}\) N & \multirow{2}{*}{4.5 km, SW} & \multirow{2}{*}{8.4} & \multirow{2}{*}{590} \\  & & 21\({}^{\circ}\)58\({}^{\prime}\)44.2\({}^{\prime\prime}\) E & & & \\  & & 49\({}^{\circ}\)59\({}^{\prime}\)42.6\({}^{\prime\prime}\) N & \multirow{2}{*}{17.2 km, E} & \multirow{2}{*}{8.7} & \multirow{2}{*}{670} \\  & & 19\({}^{\circ}\)59\({}^{\prime}\)23.2\({}^{\prime\prime}\) E & & & \\  & & 50\({}^{\circ}\)6\({}^{\prime}\)10.8\({}^{\prime\prime}\) N & \multirow{2}{*}{35.8 km, SE} & \multirow{2}{*}{8.2} & \multirow{2}{*}{700} \\  & & 19\({}^{\circ}\)27.098\({}^{\prime\prime}\) E & & & \\ \hline \hline \end{tabular}
\end{table}
Table 1: The locations and coordinates of urban experimental apiaries and the distance from them of control apiaries located in a rural area.  
We added open-mated sister queens belonging to the _Apis mellifera carica_ subspecies, born in 2020, to the bee colonies. This subspecies is native to southeastern Poland. If any queen died during the experiment, a new one was introduced, unless it was another young queen because of supersedure. The establishment of colonies commenced in June, with the utilization of approximately 1.5 kg of packed bees. During the broodless period, treatment against _Varroa_ mites was implemented using amitraz. In subsequent years, treatment with amitraz strips was scheduled to commence in August. The bees were kept in wooden Dadant hives with a frame size of 300 mm in height and 435 mm in width. The urban apiary in Lublin was located on the rooftop of a five-floor building, while the others were located on the ground level. In 2021, 84 colonies were tested (40 in the city and 44 in the countryside), and in 2022, 92 colonies were tested (49 in the city and 43 in the countryside).