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Pol. J. Environ. Stud. Vol. 27, No. 2 (2018), 787-792 DOI: 10.15244/pjoes/74154 ONLINE PUBLICATION DATE: 2018-01-10 Original Research The Impact of Selected Pesticides on Honey Bees Paweł Migdał1*, Adam Roman1, Ewa Popiela-Pleban1, Monika Kowalska-Góralska2, Sebastian Opaliński1 Department of Environment, Hygiene and Animal Welfare, Wrocław University of Environmental and Life Sciences, 51-630 Wrocław, Poland 2 Institute of Biology, Wrocław University of Environmental and Life Sciences, 51-630 Wrocław, Poland 1 Received: 2 March 2017 Accepted: 28 May 2017 Abstract The presence of honey bee in the environment has a significant impact on global agricultural production. While searching for pollen and nectar, the bee is often exposed to several contaminants such as plant protection products. The aim of this study was to assess the impact of selected pesticides from various groups (fungicides, herbicides, insecticides) on the way the bees harvested food as well as on their behavior, survival rate and the concentration of certain elements in the organisms of bee. The concentration possibly similar to this applied in agriculture and recommended by the manufacturers, were used. The research material was analyzed in terms of the presence of trace elements (manganese, copper, iron, nickel, zinc, lead, cadmium) of different toxicity. The study clearly indicates that selected pesticides used for the purpose of plant protection and plant cultivation may adversely on bees’ behavior and accumulation of trace elements in their body. Among the tested pesticides particular attention should be paid to fungicides that are not commonly tested for their effects on the honey organ. In own studies showed, their effect significantly changed the content of copper and negatively affected the survival of the bees. Keywords: behavior, trace elements, pesticides, honey bee Introduction Intense development of agriculture and animal production has caused exposure to substances with which bees have never before come into contact. The increasing demand for food has forced farmers to use more mineral fertilizers and pesticides to generate higher yields [1]. The *e-mail: pawel.migdal@upwr.edu.pl residues of these substances in the form of contaminants are then transferred into grains, vegetables, and fruit [2]. They have also been discovered in herbs such as mint (Mentha) or lemon balm (Melissa officinalis) [3]. While working on flowers, bees are exposed to direct and indirect contact with pesticides which, depending on the mode of action and the concentration of active substance, can lead to sudden death of pollinating insects or cause death within a couple of hours following exposure [4]. It becomes dangerous when the level of pesticides or their residues in a beehive becomes 788 high enough to adversely affect the functioning and development of larvae [5]. This causes the bees to become more susceptible to bacterial infections. This is particularly risky in the case of American foulbrood caused by Paenibacillus larvae. Weakened bees and a disorganized colony become more susceptible to this disease [6]. They also suffer more frequently from nosemosis, also known as bee dysentery, or varroosis [7]. Moreover, the microflora of bee intestines is affected [8]. Some pesticides trigger disorders of the digestive and endocrine systems, leading to distention of the abdomen, as a result of which the bees become slow and apathetic, showing difficulties with active flying [9]. Bees are highly susceptible to environmental changes and pollution, which is strongly reflected in the significant decrease of their survival rate [10]. The search for reasons for this phenomenon has established a new disease entity called colony collapse disorder (CCD). Several studies have indicated the important role of pesticides in the appearance and development of numerous pathological disorders in bee organisms [11]. Although it is still hard to find one cause of CCD, the significant impact of pesticides on its development cannot be questioned. Animals and their produce may also be used in the process of evaluating the level of environmental pollution. Honey bees (A. mellifera) serve as a bioindicator of contamination with trace elements of toxic properties (copper, zinc, iron, tin cadmium, lead, arsenic, and aluminum) [12]. Bees constantly penetrate the environment seeking new produce and the raw materials necessary to provide for the functioning of their colony. Even though the aforementioned elements may appear in pesticides commonly used in agriculture, it is impossible to determine unequivocally whether their presence in bees and bee products originates in the environment or results from the contact between bees and pesticides. Selected manganese (Mn) negatively affects behavior and leads to disturbances in chitin synthesis, while copper (Cu) is a natural component of the hemolymph of bees [13-14]. Lead (Pb) and cadmium (Cd) are very toxic for animals and the environment [15]. Iron (Fe), nickel (Ni), and zinc (Zn) contaminants in the honey bee body can occur because of botanical origin, as well as anthropogenic factors around the colonies [16]. The scientific literature indicates that Pb, Cd, Cu, Mn, Ni, Zn, and Fe are the most important heavy metals, and studies show a tendency for their accumulation in living organisms [17-18]. Pesticides that contain metals as active substances influence the content of these elements in the organism of honey bees. The aim of this study was to assess the impact of selected pesticides from various groups (fungicides, herbicides, insecticides) on bee behavior, survival rate, and the concentration of selected trace elements in their organisms. Migdał P., et al. Material and Methods Laboratory tests were carried out from June to midAugust. The studied material consisted of honey bee workers (A. mellifera) of the Carniola race obtained from one bee colony to each repeat [19]. Ten-day-old worker bees were distributed into seven dietary treatments. They were transported to the laboratory where the container was left for 30 min (at about 26ºC). The bees were then rendered unconscious by exposure to ammonium nitrate. After about 30 seconds, the insects became unconscious for about 2-3 min, when they were moved to experimental cages at 160 individuals per cage. The cages were supplied with lids, each with two holes with 5 cm3 food dispensers containing test or control solutions. Bees were placed in identical cages of wood and glass with dimensions of 50 × 150 × 150 mm. Inside each cage were placed frames with wax foundations (size approximately 120 × 120 mm) [20]. The cages were put in an incubator where constant temperature and humidity (the first 24 hours of experience T 35°C, subsequently 27ºC, H 75%) were maintained [21]. Throughout the study, bees were fed with 2 mol/dm3 sugar syrup. The first 24 h were dedicated to adjusting to the new environmental conditions, feeding, and then – after removal of dead individuals – starting the actual experiment. Each dietary treatment was divided into six replicates. Pesticides were selected in the manner allowing for double representation of each group (two kinds of insecticides, two herbicides, and two fungicides). The concentration of each pesticide was in accordance with the recommendation of the manufacturer. Each dose of pesticide was dissolved in 100 cm3 of sugar syrup (2 moll /dm3). Experimental groups were: –– Group K: control, pesticide-free sugar syrup. –– Group A: Miedzian 50WP (fungicide 1) with active substance of copper oxychloride- authorized for trade and distribution until 2019, dose 0.35 g/100 cm3 of syrup. –– Group B: thiram granuflo 80WG (fungicide 2) main active ingredient thiram, authorized for use until 2020, dose 0.60 g/100 cm3 of syrup. –– Group C: Fastac 100EC (insecticide1) active substance α-cyphermetrin, authorized for distribution until 2020, dose 0.04 cm3/100 cm3 of syrup. –– Group D: Actara 25WG (insecticide 2) active substance thiamethoxam, authorized for trade until 2021, dose 0.04 g/100 cm3 of syrup. –– Group E: Basagran 480SL (herbicide 1) active substance bentazone, authorized for distribution until 2020, dose 1.2 cm3/100 cm3 of syrup. –– Group F: Metafol 700SC (herbicide 2) active substance metamitron, authorized for production until 2016, dose 0.60 cm3/100 cm3 of syrup. Every day the solutions in food dispensers were replaced with new ones in order to minimize the risk of chemical changes in the tested substances. Dead individuals were collected and stored in a freezer (-20ºC). After completion of the experiment the samples The Impact of Selected Pesticides... were defrosted and triturated using a ceramic crucible. Afterward the samples were transferred quantitatively to a petri dish and placed in a dryer laboratory for 8 h at 45ºC. In order to obtain precisely dried and homogenized samples, dried material was triturated and placed on RADWAG WPX 50S moisture balances, where it was dried to a constant weight at the same temperature as in the incubator. Each sample was weighed at 1 g (to the nearest 0.10 mg) and samples of biological material were weighed in Teflon dishes using a RADWAG WAS 220/X analytical balance. The samples were covered with 5 cm3 of spectrally pure 69% nitric acid (TRACEPUR EMD Millipore Corporation). These sample preparations were mineralized in ANTON PAAR MULTIWAVE 3000 microwave digestion for 30 minutes. The whole process lasted two hours. After mineralization the obtained mineralisates were centrifuged and the content was moved into new Falcon tubes. The resulting mineralisates were analyzed quantitatively using flame atomic absorption spectrophotometry (AAS). Elements to which bees might be exposed in the natural environment and elements that appeared in the chemical structure of active substances of the pesticides were indicated. The indicated trace elements included Cu, Mn, Fe, Ni, Cd, Pb, and Zn. Moreover, behavioral observation was carried out during the study. The results obtained in experimental groups were related to a control group. The following behavioral factors were taken into account: aggression, walking, grooming, reversal, immobility, and attempts to reconstruct comb foundations. If no irregularities were noticed during observation it was marked with ‘-’ in the table, whereas any behavioral changes or dysfunctions were marked with a ‘+’ (small), ‘++’ (medium), or ‘+++’ (big). An assessment was made based on frequency of given behavior. Observations were carried out in 15-min periods for each group by two observers. The results obtained were analyzed statistically using Statistica for Windows v. 10.0. The mean, standard deviation, and relevance of differences between given groups were established. Statistical analysis was made based on Duncan test (P
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Running Head: PESTCIDE IMPACTS

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Pesticide impacts on pollinators
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PESTCIDE IMPACTS

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The journal article has insightful information that can interest any scholar regarding the
content of copper in fungicides that can harm pollinators such as bees. Since the study found out
that copper was the primary trace in affected bees, then it becomes imperative to explore copper
content...


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