Oxidative Stress Response of Honey Bee Colonies (Apis mellifera L.) during Long-Term Exposure at a Frequency of 900 MHz under Field Conditions.

In this study, oxidative stress and lipid peroxidation in honey bee larvae, pupae and the midguts of adult bees were investigated during a one-year exposure to radiofrequency electromagnetic fields (RF-EMFs) at a frequency of 900 MHz under field conditions. The experiment was carried out on honey bee colonies at three locations with electric field levels of 30 mV m-1, 70 mV m-1 and 1000 mV m-1. Antioxidant enzymes, glutathione-S-transferase (GST), catalase (CAT) and superoxide dismutase (SOD) and thiobarbituric acid reactive substances (TBARS) as indicators of lipid peroxidation were measured spectrophotometrically. The GST activity within the same developmental stage showed no significant differences regardless of electric field level or sampling time. The highest GST activity was found in the pupae, followed by activity in the larvae and midguts. Both CAT activity and TBARS concentration were the highest in the midguts, regardless of field level and sampling time. The larvae showed a significantly higher TBARS concentration at the location with an electric field level of 1000 mV m-1 compared to the locations with lower levels. Our results show that RF-EMFs at a frequency of 900 MHz can cause oxidative stress in honey bees, with the larval stage being more sensitive than the pupal stage, but there was no linear relationship between electric field level and effect in any of the developmental stages.

The Content of Cobalt, Silver and Vanadium in Raw Cow's Milk in Croatia and Risk Assessment for Consumers.

Concentrations of selected trace elements Ag, Co and V in raw milk sampled from four geographical regions in Croatia were measured. Silver, Co and V were detected above the limit of detection within the range of 9.52%-30.8%, 1.6%-12.1% and 12.4%-30.8%. Silver concentrations were not detected in milk samples from the Croatian Littoral and Mountainous Croatia (CL-MC) region. Similar Ag content was found in Southern, Eastern and Central Croatia. The lowest mean of Co and V of 33.2 and 83.8 µg kg-1 were found in the CL-MC region while the highest of 49.8 and 136.9 µg kg-1 was found in Central Croatia. There were no statistically significant differences in Ag, Co and V contents between the four regions. The estimated daily dietary intakes (EDI) of total mean and total 95th percentile values of Ag, Co and V showed lower values in comparison with available EFSA health-based limits.

Nesting behaviour and foraging characteristics of Andrena cineraria (Hymenoptera: Andrenidae).

The current studies were carried out in the three experimental locations of Kashmir valley during 2013 to 2016. The species Andrena cineraria formed the dense nest aggregations in plan grounds, barren lands and hilly areas near the fruit orchards and other landscapes with clay loam soil type. The species start flying and foraging in the orchards from April till July. The nests were allodalous, 29-36 cm in depth, with cells located obliquely around the main barrow. The nests were dense with a maximum density of 11.09 nests/m2 observed in landscapes of Budgam. The barrow diameters were found varying with depth from main entrance. The maximum barrow diameter recorded was 2.05 mm. At certain depth, the female constructs the first cell and the upper nest burrow is vertical and lower is oblique. The nest entrance is generally hidden under the tumulus. In the depth of average 30.48 cm, each cell directly opens to main burrow either alternately or unilaterally. The cell number, diameter, and length varied with depth. Foraging behaviour of A. cineraria on various fruit crops and other shrubs and social forestry trees were determined and the abundance, visitation rate, total visits and time spend per flower were found significant, especially on fruit crops. The significance of the studies is important for the melittologists, as it will help in the conservation of bee fauna. The study is also important in using this species for pollination purpose and would also help to detect and understand the possible pre-adaptation of species in temperate region of Kashmir valley.

Mating behaviour and behavioural ecology of a Predatory Wasp, Symmorphus allobrogus (de Saussure) (Hymenoptera: Eumeninae).

This paper represents an attempt to investigate the mating behaviour of Symmorphus allobrogus, explaining the willingness of male to mount and copulate. The male displays including mode and frequency of antennation and position while copulating, the displays further comprises of intensity and frequency of rejecting behaviour. The presence of the male's copulatory and postcopulatory courtship studies, understands the maintenance of monandry. The wasp has numerous secondary sexual characters, and the mating behaviour follows a phyletic and the specific sexual mating characters in context of sexual selection. The duration of mating phases and the number of male antennation series during precopulatory, copulatory and postcopulatory phases of mounting, differs significantly. Mating success depends mostly on the activities of male in the premounting phase and the behaviour of both sexes has a roughly equal importance for it in precopulatory phase. While during copulation, activity of male has little influence on its duration; however, behaviour of female has crucial effect, inducing its earlier termination.

An international inter-laboratory study on Nosema spp. spore detection and quantification through microscopic examination of crushed honey bee abdomens.

Nosemosis is a microsporidian disease causing mortality and weakening of honey bee colonies, especially in the event of co-exposure to other sources of stress. As a result, the disease is regulated in some countries. Reliable and harmonised diagnosis is crucial to ensure the quality of surveillance and research results. For this reason, the first European Interlaboratory Comparison (ILC) was organised in 2017 in order to assess both the methods and the results obtained by National Reference Laboratories (NRLs) in counting Nosema spp. spores by microscopy. Implementing their own routine conditions of analysis, the 23 participants were asked to perform an assay on a panel of ten positive and negative samples of crushed honey bee abdomens. They were asked to report results from a qualitative and quantitative standpoint. The assessment covered specificity, sensitivity, trueness and precision. Quantitative results were analysed in compliance with international standards NF ISO 13528 (2015) and NF ISO 5725-2 (1994). Three results showed a lack of precision and five a lack of trueness. However, overall results indicated a global specificity of 98% and a global sensitivity of 100%, thus demonstrating the advanced performance of the microscopic methods applied to Nosema spores by the NRLs. Therefore, the study concluded that using microscopy to detect and quantify spores of Nosema spp. was reliable and valid.

Mineral Content in Honeybee Wax Combs as a Measurement of the Impact of Environmental Factors.

Environmental pollution from metals needs to be constantly monitored due to their predominantly negative impacts on living organisms. As apian products stored in hives are considered useful bioindicators, the objective of this study was to: (a) investigate and compare the essential and toxic metal concentrations in freshly constructed combs (light combs, LC) and old combs (dark combs, DC) in use for two to three beekeeping seasons, and (b) compare the mineral content of beeswax combs from apiaries exposed to different levels of environmental pollution using the energy dispersive x-ray fluorescence method. Concentrations of ten elements (Cr, Pb, Cu, Ni, Fe, Zn, Mn, Sr, Rb, Ca) were determined in 18 honeybee wax comb samples from three apiaries in continental Croatia. The results showed that the influence of comb age and/or geographical origin (representing varying levels of environmental pollution exposure) on the elemental composition of beeswax was evident for the toxic elements Cr, Pb, Cu and Ni, and for the essential elements Fe, Zn, Mn and Sr, but not Rb. In addition to monitoring the environmental element content, wax combs can be used to determine contamination levels. Additionally, in-time analysis results can enable beekeepers to adjust management practices, such as moving apiaries to better positions. They can also be useful in the creation of policies on acceptable limits for toxic metal levels in particular geographical areas.

Trueness and precision of the real-time RT-PCR method for quantifying the chronic bee paralysis virus genome in bee homogenates evaluated by a comparative inter-laboratory study.

The Chronic bee paralysis virus (CBPV) is the aetiological agent of chronic bee paralysis, a contagious disease associated with nervous disorders in adult honeybees leading to massive mortalities in front of the hives. Some of the clinical signs frequently reported, such as trembling, may be confused with intoxication syndromes. Therefore, laboratory diagnosis using real-time PCR to quantify CBPV loads is used to confirm disease. Clinical signs of chronic paralysis are usually associated with viral loads higher than 108 copies of CBPV genome copies per bee (8 log10 CBPV/bee). This threshold is used by the European Union Reference Laboratory for Bee Health to diagnose the disease. In 2015, the accuracy of measurements of three CBPV loads (5, 8 and 9 log10 CBPV/bee) was assessed through an inter-laboratory study. Twenty-one participants, including 16 European National Reference Laboratories, received 13 homogenates of CBPV-infected bees adjusted to the three loads. Participants were requested to use the method usually employed for routine diagnosis. The quantitative results (n=270) were analysed according to international standards NF ISO 13528 (2015) and NF ISO 5725-2 (1994). The standard deviations of measurement reproducibility (SR) were 0.83, 1.06 and 1.16 at viral loads 5, 8 and 9 log10 CBPV/bee, respectively. The inter-laboratory confidence of viral quantification (+/- 1.96SR) at the diagnostic threshold (8 log10 CBPV/bee) was+/- 2.08 log10 CBPV/bee. These results highlight the need to take into account the confidence of measurements in epidemiological studies using results from different laboratories. Considering this confidence, viral loads over 6 log10 CBPV/bee may be considered to indicate probable cases of chronic paralysis.

Impact of Thiamethoxam on Honey Bee Queen (Apis mellifera carnica) Reproductive Morphology and Physiology.

High honey bee losses around the world have been linked in part by the regular use of neonicotinoids in agriculture. In light of the current situation, the aim of this study was to investigate the effects of thiamethoxam on the development of the reproductive system and physiology in the honey bee queen. Two experimental groups of honey bee queen larvae were treated with thiamethoxam during artificial rearing, applied via artificial feed in two cycles. In the first rearing cycle, honey bee larvae received a single treatment dose (4.28 ng thiamethoxam/queen larva on the 4th day after larvae grafting in artificial queen cells), while the second honey bee queen rearing cycle received a double treatment dose (total of 8.56 ng thiamethoxam/queen larva on the 4th and 5th day after larvae grafting in artificial queen cells). After emerging, queens were anesthetized and weighed, and after mating with drones were anesthetized, weighed, and sectioned. Ovary mass and number of stored sperm were determined. Body weight differed between untreated and treated honey bee queens. The results also show a decrease in the number of sperm within honey bee queen spermathecae that received the double thiamethoxam dose.