# Honey as a Bioindicator: Pollution's Effects on Its Quality in Mining vs. Protected Sites
## 2 Materials and Methods  
### Honey Samples and the Description of the Area  
In July 2024, 1-2 kg of polyfloral honey (with a moisture content of approximately 16-18% at the time of sampling) was collected from each apiary (from about 100 hives per apiary), out of the total amount harvested by beekeepers. A total of 8 honey samples were collected, including 4 composite honey samples from mining areas (Table 1) and 4 composite samples from adjacent protected areas (Table 2). The composite samples consisted of homogenized honey from the same apiary, originating from approximately 100 beehives. Each composite sample was divided into 3 subsamples, so that each could be analyzed in triplicate. The samples were collected from plastic containers in which the honey was stored. They were then placed in glass jars and kept at a temperature of approximately 2-8 \({}^{\circ}\)C until analysis. \begin{table}
\begin{tabular}{c c c c} \hline \hline
**Sample No.** & **County** & **Area** & **Mineral Resource Type** \\ \hline S1 & Alba & Zlatna Mining Operation, & Gold, silver, copper, lead, \\  & & Hanes Mine & and zinc. \\ S2 & Hunedoara & Certeju de Sus Mining & Gold, silver, copper, lead, \\  & & Operation & and zinc. \\ S3 & Bihor & Baita Mining Operation & Gold, silver, copper, lead, \\  & & & zinc, and uranium. \\ S4 & Harghita & Baían Mining Operation & Copper, gold, silver, lead, \\  & & & and zinc. \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Identification of honey samples from locations associated with mining operations in Romania. \begin{table}
\begin{tabular}{c c c c} \hline \hline
**Sample No.** & **County** & **Area** & **Biodiversity Protection Site** \\ \hline SS1 & Alba & Almasu Mare Commune & ROSCI0029 Glodului, Cibului, and Mizii Gorges \\ SS2 & Hunedoara & Balsa Commune & ROSPA0132 Metaliferi \\  & & Mountains \\ SS3 & Bihor & Cristioru de Jos & ROSCI0324 Bihor Mountains \\ SS4 & Harghita & Baían Town & ROSAC0027 Bicaz–Hásmas \\  & & & Gorges \\ \hline \hline \end{tabular}
\end{table} 
Table 2: Identification of honey samples collected from protected natural areas in Romania. Table 1 presents the identification of the honey samples collected from areas located in the vicinity of former mining sites in Romania, highlighting the geographical location and the types of mineral resources associated with each area. Table 2 includes the identification of the honey samples collected from protected natural areas in Romania, indicating the administrative location and the sites of community or avifaunal importance where each apiary's area is located. Figure 1 presents a map of Romania showing the locations where the honey samples were collected, both from areas near mining operations and from protected natural zones, highlighting the geographical distribution of the apiaries analyzed in the study. 1. The Zlatna Mining Operation--Hanes Mine (Alba County), located in the Apuseni Mountains, has a millennia-long mining tradition. Industrial exploitation began in the 18 th century under Austrian administration, focusing on polymetallic ones--mainly copper, as well as gold, silver, lead, and zinc. The mine expanded significantly during the communist era, becoming part of the Zlatna Mining Complex after nationalization in 1948. Operations declined in the 1990s, due to declining resources, high operational costs, and environmental issues, and it was officially closed in 2003. The area has undergone ecological rehabilitation, through heavy metal pollution and abandoned infrastructure remain [22; 23]. 2. The Certeju de Sus Mining Operation (Hunedoara County), part of the gold-rich Metalifera Mountains, was industrially developed starting in the 19 th century and intensified under communism. The mine produced complex ores rich in gold and silver, as well as copper, lead, and zinc. Its mineralization is characterized by the presence of auriferous pyrites, commonly associated with galena, sphalerite, chalcopyrite, and gold tellurides.