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Chalkbrood is a disease of honey bee brood caused by the fungal parasite Ascosphaera apis. Many factors such as genetics, temperature, humidity and nutrition influence the appearance of clinical symptoms. Poor nutrition impairs the immune system, which favors the manifestation of symptoms of many honey bee diseases. However, a direct link between dietary ingredients and the symptoms of chalkbrood disease has not yet been established. We show here that the elemental composition of chalkbrood mummies and healthy larvae from the same infected hives differ, as well as that mummies differ from larvae from healthy hives. Chalkbrood mummies had the highest concentration of macroelements such as Na, Mg, P, S, K and Ca and some microelements such as Rb and Sn, and at the same time the lowest concentration of B, As, Sr, Ag, Cd, Sb, Ba and Pb. Larvae from infected hives contained less Pb, Ba, Cs, Sb, Cd, Sr, As, Zn, Cu, Ni, Co, Mn, Cr, V and Al in contrast to healthy larvae from a disease-free apiary. This is the first study to demonstrate such differences, suggesting that an infection alters the larval nutrition or that nutrition is a predisposition for the outbreak of a chalkbrood infection. Though, based on results obtained from a case study, rather than from a controlled experiment, our findings stress the differences in elements of healthy versus diseased honey bee larvae.

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Honey bees are social insects that show division of labor and sexual dimorphism. Female honey bees differentiate in two different castes, queens or worker bees, while males are called drones. Worker bees have different tasks in the hive including collection of food, its processing, caring for brood, protecting the hive, or producing wax. The drones' only role is to mate with a virgin queen. Many studies have dealt with differences in physiology, behavior, and morphology of workers and drones. This is the first study that demonstrates differences in element accumulation and composition between workers and drones honey bees. Using inductively coupled plasma mass spectrometry, we found that worker honey bees have higher concentrations of most elements analyzed. Drones had higher concentrations of elements essential to bees, Na, P, S, Zn, Cu, and especially Se (2.2 × higher), which is known to be important for sperm quality and fertility in many animals. Until now higher Se content was not observed in male insects. These differences can be attributed to different environmental exposure, reproductive role of drones, but mostly to the food workers and drones consume. Worker bees feed on bee bread, which is rich in minerals. Drones are fed food pre-processed by worker bees.

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Arsenic can be toxic to living organisms, depending not only on the concentration, but also its chemical form. The aim of this study was to determine arsenic concentrations and perform arsenic speciation analysis for the first time in honeybees, to evaluate their potential as biomonitors. Highest arsenic concentrations were determined in the vicinity of coal fired thermal power plants (367 µg kg-1), followed by an urban region (213 µg kg-1), with much lower concentrations in an industrial city (28.8 µg kg-1) and rural areas (41 µg kg-1). Until now, honey bees have never been used to study different arsenic species in the environment. For this reason, four extraction procedures were tested: water, hot water at 90 °C, 20% methanol, and 1% formic acid. Water at 90 °C was able to extract more than 90% of the total arsenic from honey bee samples. Inorganic arsenic (the sum of arsenite and arsenate) accounted for 95% of arsenic species in bees from three locations, except the industrial city, where it represented only 80% of arsenic species, while 15% was present as DMA.

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Homogenized or pooled samples of honey bees are already used for monitoring of metal pollution in the environment for a couple of decades. This is the first study that analyzed the elemental composition of individual honey bees. One apiary with 21 hives was used to test in-hive and between-hive differences at the same location. Highest in-hive variability was observed for Al (14.1x), Li (10.3x), V (10.3x), As (8.9x) and Cd (7.9x). For the elements Cu, K, Mg, Na, P, S and Zn the smallest both in-hive (2.3x, 1.8x, 2.0x, 2.2x, 1.9x, 1.7x, 2.4x respectively) as well as between-hive variability (1.5x, 1.4x, 1.4x, 1.6x, 1.4x, 1.4x and 1.5x respectively) was observed. Nonetheless, between-hive variability was statistically significant for all the analyzed elements. We proved that these differences significantly influence comparison of element concentrations in bees from different locations. One hive from Mesic apiary had significantly lower concentrations of Al (hive 18: 18.6 mg kg-1) and U (hive 1: 0.0013 mg kg-1), while at the same time a different hive from the same apiary showed higher concentration of these elements (Al, hive 14: 125 mg kg-1; U, hive 13: 0.012 mg kg-1) compared to another location (thermal power plant, Al: 97 mg kg-1, U: 0.0044 mg kg-1). Therefore, when using honey bees as biomonitors, we recommend a larger number of hives to be sampled at each location to provide an adequate dataset for reliable interpretation of results. Comparing individual bee elemental concentrations, for Na, Mg, P, S, K, Fe, Cu, and Zn negative correlations were found between dry mass of a bee and concentrations of these elements. These negative correlations could be a consequence of higher honey or nectar content in some of the sampled bees. However, this should also be considered when using honey bees as bioindicators.

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Honeybee colony losses have been a focus of research in the last years, due to the importance of managed honeybee colonies for economy and ecology. Different unfavorable conditions from the outside environment have a strong impact on the hive health. The majority of losses occur mainly during winter and the exact reason is not completely understood. Only a small number of studies are dealing with content of bioelements, their function and influence on honeybee physiology. The aim of the present study was to determine seasonal and spatial variations in content of bioelements and non-essential elements, in hemolymph and whole body of honeybees originating from three regions with different degrees of urbanization and industrialization. Concentrations of 16 elements were compared: macroelements (Ca, K, Mg, Na), microelements (Cu, Fe, Mn, Zn) and non-essential elements (Al, Ba, Cd, Co, Cr, Ni, Pb, Sr) in samples collected from 3 different environments: Golija (rural region), Belgrade (urban region) and Zajaca (industrial region). Content of bioelements and non-essential elements in honeybees was under noticeable influence of the surrounding environment, season and degree of honeybee activity. Hemolymph was proven to be helpful in differentiating air pollution from other sources of honeybee exposure. The results of our study demonstrated that bees can be successfully used as biomonitors since we have observed statistically significant differences among observed locations, but unless compared locations are exposed to excessively different pollution pressures, it is essential that all bees should be collected at the same season.

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With the increase in anthropogenic activities metal pollution is also increased and needs to be closely monitored. In this study honeybees were used as bioindicators to monitor metal pollution. Metal pollution in honeybees represents pollution present in air, water and soil. Concentrations of As, Cs, Hg, Mo, Sb, Se, U and V were measured. The aim of this study was to assess spatial and temporal variations of metal concentrations in honeybees. Samples of honeybees were taken at five different regions in Serbia (Belgrade - BG, Pancevo - PA, Pavlis - PV, Mesic - MS, and Kostolac - TPP) during 2014. Spatial variations were observed for Sb, which had higher concentrations in BG compared to all other regions, and for U, with higher concentrations in the TPP region. High concentrations of Sb in BG were attributed to intense traffic, while higher U concentrations in the TPP region are due to the vicinity of coal fired power plants. In order to assess temporal variations at two locations (PA and PV) samples were taken during July and September of 2014 and June, July, August and September of 2015. During 2014 observing months of sampling higher concentrations in July were detected for Sb and U in BG, which is attributed to lifecycle of plants and honeybees. During the same year higher concentrations in September were observed for As, Sb in PA and Hg in PV. This is due to high precipitation during the peak of bee activity in spring/summer of 2014. No differences between months of sampling were detected during 2015. Between 2014 and 2015 statistically significant differences were observed for Hg, Mo and V; all elements had higher concentrations in 2014. This is in accordance with the trend of reduction of metal concentrations in the bodies of honeybees throughout the years in this region.

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In this study, honeybees were used to determine spatio-temporal variations and origin sources of Pb. Lead concentrations and isotopic composition were used in combination with selected statistical methods. The sampling was carried out at five different locations in Serbia: urban region (BG), petrochemical industry (PA), suburban region (PV), rural region (MS) and thermal power plant region (TPP) during 2014. At PA and PV locations, samples were taken during multiple years. This is the first use of Kohonen self-organizing map (SOM) in combination with honeybees as bioindicators to determine spatio-temporal variations and origin of Pb pollution. It was observed that during the years Pb concentrations were in decline. Anthropogenic sources are most dominant in BG and TPP, in PA there are mixed sources of natural and anthropogenic origin and in PV Pb is of natural origin. It can be concluded that honeybees in combination with SOM can be used to differentiate between slight changes in spatio-temporal variations of Pb, as well as for source appointment.

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The ability of honeybees to collect particulate matter (PM) on their bodies makes them outstanding bioindicators. In this study, two cities, Pancevo (PA) and Vrsac (VS), South Banat district, Vojvodina, Serbia, were covered with two sampling sites each. The aims of this study were to determine concentrations of Al, Ba, Cd, Co, Cr, Cu, Fe, Mn, Na, Ni, Sr, and Zn in the bodies of honeybees during July and September of 2013, 2014, and 2015 and to analyze their spatial and temporal variations and sources of analyzed elements, as well as to assess pollution levels in the two cities. Significant temporal differences were found for Al, Ba, Cd, Co, Cr, Cu, Fe, Mn, Na, Ni, and Zn. Trend of reduction in metal concentrations in bodies of honeybees during the years was observed. Statistically significant spatial variations were observed for Al, Ba, and Sr, with higher concentrations in VS. PCA and CA analyses were used for the first time to assess sources of metals found in honeybees. These analyses showed two sources of metals. Co, Cd, Na, Fe, Mn, Zn, and partly Cu were contributed to anthropogenic sources, while Ca, Al, Mg, Cr, Ba, Sr, and Ni were contributed to natural sources.

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BACKGROUND: Stigmatization of psychiatric patients is present both in the general population and among healthcare professionals. AIM: To determine the attitudes and behaviour of medical students towards a person who goes to a psychiatrist, before and after psychiatric rotation, and to compare those attitudes between medical and non-medical students. METHODS: The study included 525 medical students (second and sixth year of studies) and 154 students of law. The study instrument was a three-part self-reported questionnaire (socio-demographic data, Rosenberg Self-Esteem Scale and a vignette depicting a young, mentally healthy person). The experimental intervention consisted of ascribing a 'psychiatric label' to only one set of vignettes. All the vignettes (with or without the 'psychiatric label') were followed by 14 statements addressing the acceptance of a person described by vignette, as judged by social distance (four-point Likert scale). RESULTS: Higher tendency to stigmatize was found in medical students in the final year, after psychiatric rotation (Z(U) = -3.12, p = .002), particularly in a closer relationship (Z(U) = -2.67, p = .007) between a student and a hypothetical person who goes to a psychiatrist. The non-medical students had a similar tendency to stigmatize as medical students before psychiatric rotation (Z(U) = -0.03, p = .975). Neither gender, nor the size of student's place of origin or average academic mark was associated with the tendency to stigmatize in our sample. However, student's elf-esteem was lower in those with a tendency to stigmatize more in a distant relationship (ρ = -0.157, p = .005). CONCLUSIONS: Psychiatric education can either reinforce stigmatization or reduce it. Therefore, detailed analyses of educational domains that reinforce stigma will be the starting point for anti-stigma action.

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