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Background With numerous endemic subspecies representing four of its five evolutionary lineages, Europe holds a large fraction of Apis mellifera genetic diversity. This diversity and the natural distribution range have been altered by anthropogenic factors. The conservation of this natural heritage relies on the availability of accurate tools for subspecies diagnosis. Based on pool-sequence data from 2145 worker bees representing 22 populations sampled across Europe, we employed two highly discriminative approaches (PCA and F-ST) to select the most informative SNPs for ancestry inference. Results Using a supervised machine learning (ML) approach and a set of 3896 genotyped individuals, we could show that the 4094 selected single nucleotide polymorphisms (SNPs) provide an accurate prediction of ancestry inference in European honey bees. The best ML model was Linear Support Vector Classifier (Linear SVC) which correctly assigned most individuals to one of the 14 subspecies or different genetic origins with a mean accuracy of 96.2% +/- 0.8 SD. A total of 3.8% of test individuals were misclassified, most probably due to limited differentiation between the subspecies caused by close geographical proximity, or human interference of genetic integrity of reference subspecies, or a combination thereof. Conclusions The diagnostic tool presented here will contribute to a sustainable conservation and support breeding activities in order to preserve the genetic heritage of European honey bees. European CommissionEuropean CommissionEuropean Commission Joint Research Centre [2013.1.3-02, 613960]; Basque GovernmentBasque Government [IT1233-19] The SmartBees project was funded by the European Commission under its FP7 KBBE programme (2013.1.3-02, SmartBees Grant Agreement number 613960) https://ec.europa.eu/research/fp7.MP was supported by a Basque Government grant (IT1233-19). The funders provided the financial support to the research, but had no role in the design of the study, analysis, interpretations of data and in writing the manuscript. WOS:000614429400001 2-s2.0-85100413967 PubMed: 33535965

Simple Summary The miteVarroa destructorrepresents a great threat to honey bees and the beekeeping industry. The opportunity to select and breed honey bees that are naturally able to fight the mite stands a sustainable solution. This can be achieved by evaluation of the failure of mite reproduction (SMR, suppressed mite reproduction). We conducted a large European experiment to assess the SMR trait in different populations of honey bees spread over 13 different countries, and representing different honey bee populations. The first goal was to standardize and validate the SMR evaluation method, and then to compare the SMR trait between the different populations. Our results indicate that it is necessary to examine at least 35 brood cells infested by a single mite to reliably estimate the SMR score of any given colony. Several colonies from our dataset display high SMR scores, indicating that this trait is present within the European honey bee populations. No major differences could be identified between countries for a given population, or between populations in different countries. This study shows the potential to increase selection efforts to breedV. destructorhoney bee resistant populations. In the fight against theVarroa destructormite, selective breeding of honey bee (Apis melliferaL.) populations that are resistant to the parasitic mite stands as a sustainable solution. Selection initiatives indicate that using the suppressed mite reproduction (SMR) trait as a selection criterion is a suitable tool to breed such resistant bee populations. We conducted a large European experiment to evaluate the SMR trait in different populations of honey bees spread over 13 different countries, and representing different honey bee genotypes with their local mite parasites. The first goal was to standardize and validate the SMR evaluation method, and then to compare the SMR trait between the different populations. Simulation results indicate that it is necessary to examine at least 35 single-infested cells to reliably estimate the SMR score of any given colony. Several colonies from our dataset display high SMR scores indicating that this trait is present within the European honey bee populations. The trait is highly variable between colonies and some countries, but no major differences could be identified between countries for a given genotype, or between genotypes in different countries. This study shows the potential to increase selective breeding efforts ofV. destructorresistant populations.

The importance of varroosis control in a natural and sustainable way is crucial for beekeeping, having in view the varroa mite impact on honey bee health. In the last years, we developed a highly effective procedure for treating varroa in capped brood using volatile organic acids. This procedure can be applied at any moment of the active season as it uses organic substances. Taking into account the necessity to drastically reduce the level of varroa infestation in colonies before winter bee rearing, we developed a relatively simple pilot study to preliminarily test the impact of spring treatments on varroa infestation level in brood, to be evaluated in summer when, naturally, the population of mites increases. To test the hypothesis, two experimentally treated groups and a control group were used. The treatment consisted of brushing all capped brood with formic acid of 65% concentration in one and two applications. The obtained results show very significant differences between the treated and control groups in terms of infested cell percentages evaluated in the July–August period. Consequently, the spring treatments could be an important tool in limiting the varroa mite multiplication, but further experiments are necessary to test and adapt them to different local conditions.

Honeybees (Apis mellifera L.) are excellent biosensors that can be managed to collect valuable information about environmental contamination. The main objective of the present study was to design and apply an integrative protocol to monitor honeybee colony activity and sample collection by using electronic technologies combined with classical methods in order to evaluate the exposure of honeybees to the neonicotinoids that are used in melliferous intensive crops. The monitored honeybee colonies were especially prepared and equipped to maximize their chances to collect representative samples in order to express, as well as possible, the pesticide residues that existed in the targeted crops. The samples of honey, pollen and honeybees were collected, preserved and prepared to fulfill the required quality and quantity criteria of the accredited laboratories. In total, a set of fifty samples was collected from fields, located in different areas of intensive agriculture in Romania, and was analyzed for five neonicotinoids. The obtained results show that 48% of the total analyzed samples (n = 50) contained one or more detected or quantified neonicotinoid residues. The main conclusion is that the proposed approach for sample collection and preparation could improve the evaluation methodologies for analyzing honeybees&rsquo, exposure to pesticides.

Simple Summary Honey bees, in addition to producing honey, are important pollinators of wild and cultivated plants. Unfortunately, in some places, the population of honey bees is declining. One of the factors that affect their survival is adaptation to the local environment. Bees native to a particular area survive better than those imported. Despite this fact, some beekeepers import non-native bees and use them in their apiaries. Imported bees produce hybrids with bees from surrounding colonies because beekeepers do not control their mating. In consequence, the whole population can change. In this study, we verified how the population of Romanian bees has changed over the last four decades. We found significant temporal changes in wing venation. Despite these changes, the two major subpopulations of bees separated by mountains remain distinct. We provide a tool for the easy identification of native bees from Romania, which can help to protect them. Abstract The honey bee (Apis mellifera) is an ecologically and economically important species that provides pollination services to natural and agricultural systems. The biodiversity of the honey bee is being endangered by the mass import of non-native queens. In many locations, it is not clear how the local populations have been affected by hybridisation between native and non-native bees. There is especially little information about temporal changes in hybridisation. In Romania, A. m. carpatica naturally occurs, and earlier studies show that there are two subpopulations separated by the Carpathian Mountains. In this study, we investigated how the arrangement of veins in bees’ wings (venation) has changed in Romanian honey bees in the last four decades. We found that in the contemporary population of Romanian bees, there are still clear differences between the intra- and extra-Carpathian subpopulations, which indicates that natural variation among honey bees is still being preserved. We also found significant differences between bees collected before and after 2000. The observed temporal changes in wing venation are most likely caused by hybridisation between native bees and non-native bees sporadically introduced by beekeepers. In order to facilitate conservation and the monitoring of native Romanian bees, we developed a method facilitating their identification.

It is well known that breeding is based on phenotypic and behavioural performance assessed at the level of each honeybee colony. By selection, the genes responsible for the desired characters have to be favoured, by evaluation and classification of all colonies involved in a breeding program. Generally, in the beekeeping practice, the most applied method of selection is the mass selection regarding the main objective- honey production. Some more elaborated programs use selection simultaneous selection on several characters. Until now, a standard method for honey bees evaluation and selection on several characters could not be generalized, every breeder establishing the selection method depending on proposed goals which could be different especially when we speak about different races and environmental conditions. Taking into account the selection objectives in Romania it was conceived a standardized methodology for the selection on several characters in Romanian condition.

The biochemical analyses of the blood are largely used for the routine diagnosis and especially for the metabolic survey in farm animals. These facts conduct us to the idea that similar analyses, applied on honeybee hemolymph, could be used IN monitoring the healthy state of honeybee colonies. The present studies represent preliminary researches, which aimed to investigate the variability of the main biochemical parameters in the hemolymph of the healthy honeybees (Apis mellifera) in inactive season. The researches were carried out on honeybee samples collected from 5 honeybee colonies belonging to a breeding apiary of the Institute for Beekeeping Research and Development from Bucharest. In order to perform the biochemical analyses, the honeybees samples, consisting in 50 individuals on sample (10 individuals/colony) were randomly collected and their haemolimph collected, at different time intervals, in inactive season (fall-winter). Totally, there were collected 250 haemolyph samples in a 2 years interval and the following 21 biochemical parameters were analysed: GLU, HDL-c, ALP, T-cho, Tprot, Alb., BUN, LDH, CPK,, Mg, IP), GGT, GOT, GPT, Ca, Cre,, Amy, T–BIL, TG, UA.. The test was carried out after the collection and processing of the samples using the SPOTCHEM EZSP4430, equipment with dry kits, the slides technique, respectively .During the 2nd part of the inactive season, the values of most biochemical parameters increase in different proportions, their levels being maintained also in the first part of the active seasons (April, May, June).The values obtained for the main studied biochemical parameters in the haemolymph of the healthy honeybees collected from honeybee colonies kept in natural conditions show a highly variable evolution in the two consecutive years of experiments during the inactive season.

The main objective of this clinical study consisted in evaluation of the therapeutic effects of the propolis extract used in different disorders at company animals, thus being improved the palette of the apitherapeutical products used in veterinary purposes. The experiments were carried out on company animals (two experimental groups) during the 2007-2008 period, in the frame of the Veterinary Medicine Faculty – Bucharest and the University - Spiru Haret, at the veterinary departments: Parasitology, Dermatology and Surgery. The raw propolis was collected from the bee colonies belonging to the Institute of the Beekeeping Research & Development– Bucharest and the apiphytotherapeutical product based on propolis was obtained in the Apitherapy sector of the same Institute. In a first stage were obtained the antiparasite, dermatological and surgical veterinary product PROACTIVATOR based on propolis alcoholic extract and Aloe vera gel. The experiments consisted in administration of the obtained preparation in different disorders on the experimental groups as: dermatological (plagues, chemical and physical burns), parasitological (extern parasites: scabies supra infected or not) and in veterinary surgery (as a protective layer applied on the sutured plague). In dermatologic disorders the effects of the PROACTIVATOR product were established by way of clinical periodical examinations until the total recover were done. In external parasites and connected disorders it was established the repellent or killing effect of the preparation on the infestation with parasites and the degree of control in the correlated infections. In skin tissue surgery it was established the cicatrising effect in sutured plagues and the anaesthesic local effect. The established of the studied preparation efficiency was similar as those used in classical treatments with synthesis products. The advantage of the utilization of PROACTIVATOR eliminates the toxic and cumulative effects of the chemical substances as it is well known.

The international investigations regarding the honeybees’ diversity carried out until now have revealed a certain degree of genetic pollution in different countries from Europe, because of the import of more productive honeybees’ races or of some interracial honeybees’ hybrids. This fact might have a negative impact on the success adaptability of honeybees at the ecosystem. Although, the Romanian honeybees (Apis mellifera carpathica) are well adapted to the local conditions and express a good resistance to diseases, the introgression (genetic pollution) of different honeybees’ races could be an imminent event. So that, starting from 2007, by a cooperation between the Institute for Beekeeping Research and Development from Bucharest and the Institute of Genetics of the University of Bucharest, we have initiated different investigations in order to obtain a more accurate state of the Romanian honeybees’ diversity. We have performed specific molecular analyses, using mtDNA (the COI-COII test) extracted from 32 different honeybees samples collected from several regions from Romania. For a better and detailed characterization of the collected honeybee’s samples we have also carried out some morphometric measurements of their wings. Our data have shown that the Romanian population of honeybees is almost homogenous from the genetic and the morphometric points of views. These types of investigations represent a premiere for Romania.

The electronic hive is the result of the scientific researches carried out between2003-2006 by a research project funded by MEdC through the National ProgramRELANSIN, being accomplished by Institute for Beekeeping Research andDevelopment –Bucharest in cooperation with the Polytechnics University fromBucharest –The Center for Electronic Technology and Interconnection Techniquesand the Radio Consult CompanyTo achieve the great complexity of the electronic model adapted to the hive –the“smart” hive, it was necessary to establish the all electronic details which to makepossible to monitorize some very important information from the bee colony andits environment with the help of the honeybees and which to eliminate the errorsthat may occur in the information collection process.Thus, the project aimed to conceive the electronic system in order to collectinformation from inside the hive and from environment too, to storage andtransmit it to a data basis by GSM network in order to be analyzed and processedby users.By this complex electronic system, composed by electronic equipment and thehoney bee colony, which is dynamic and strong related with natural evolution ofvegetation correlated with the climate factors, is possible to identify instantaneousor periodically a large palette of aggression factors as well naturals (acids rains,extreme temperatures, calamities) as anthropic factors –accidental chemical orbiologic pollution. The obtained data, electronically quantified and taken out intothe data basis, could offer accurate information about the moisturized areas atdifferent time intervals.

Although biochemical analyses of the blood were and are still used for the routinediagnosis and especially for the metabolic survey in farm animals, such analyses, maybe applied for the honeybee hemolymph (as a paraclinic examination). The aim of thisexperimental study was to investigate and to determine the reference values of themain biochemical parameters in the hemolymph of the healthy honeybees of Apismellifera carpathica. The honeybee samples were collected in order to analyze thehealth condition of the respective colony. All the samples coming from sick colonieswere removed, only healthy adult honeybees coming from strong colonies were keptand used for hemolymph collection. By special methods, samples of hemolymph (300μl/sample) collected from about 50 individuals, were analyzed both during the activeseason (spring/summer) and the inactive season (autumn/winter). The study wascarried out on 50 samples of undiluted hemolymph taken from a total number of about2,500 honeybees. The following 21 biochemical parameters were analyzed: GLU(mg/dl), HDL-c (mg/dl), ALP (UI/l), T-cho (mg/dl), Tprot (mg/dl), Alb. (g/dl), BUN(mg/dl), LDH (UI/l), CPK (UI/l), Mg (mg/dl), FRA (μm/l), IP (mg/dl), GGT (UI/l),GOT (UI/l), GPT (UI/l), Ca (mg/dl), Cre (mg/dl), Amy (UI/l), T–BIL (mg/dl), TG(mg/dl), UA (mg/dl). The test was carried out after the collection and processing of thesamples using the SPOTCHEM EZSP4430, equipment with dry kits, the slides technique,respectively.

In the last years, in internationally context, was proved that long-termchemotherapy could have negative impact, increasing the resistance of beediseases to the medication with consequences from economic points of viewespecially on the quality of beekeeping products. In this context, the strategy ofbee diseases control was reanalyzed, the researches being reoriented throughfinding the alternative and non-polluted solutions, which could be integrated inan efficient system of bee diseases control, depending on breed, climate, andmaintaining systems. Thus, one of the research directions is the ameliorativeselection of bees for natural diseases resistance combined with other desiredmorpho-productive and behavioral characteristics, in this way a series ofspecific tests being already established. The hygienic behaviour is the primaryway to defend against some bee diseases, especially to American foulbrood(Rothenbuhler, 1964) and chalk brood (Gilliam, 1983). The honeybees with highhygienic behavior detect, uncap and remove the infested brood more rapidly thanothers, before the disease to be spread.The preliminary researches carried out inthe breeding apiaries of the Beekeeping Research and Development Institute(Bucharest) aimed to establish the natural diseases resistance degree of local beepopulations (A. m. carpathica), by applying specific tests to identify the beecolonies with high hygienic behaviour.

International audience; Here, scientists from 19 European countries, most of them collaborating in Working Group 4: “Diversity and Vitality” of COST Action FA 0803 “Prevention of honey bee COlony LOSSes” (COLOSS), review the methodology applied in each country for discriminating between honey bee populations. Morphometric analyses (classical and geometric) and different molecular markers have been applied. Even if the approach has been similar, however, different methodologies regarding measurements, landmarks or molecular markers may have been used, as well as different statistical procedures. There is therefore the necessity to establish common methods in all countries in order to have results that can be directly compared. This is one of the goals of WG4 of the COLOSS project; Científicos de 19 países europeos, la mayoría de ellos colaboradores del Grupo de Trabajo 4: "Diversidad y Vitalidad" de la Acción COST FA 0803 "Prevención de la mortalidad de colmenas" (COLOSS), revisan aquí la metodología aplicada en cada país para discriminar a las poblaciones de abejas. Se han aplicado análisis morfométricos (clásico y geométrico) y distintos marcadores moleculares. Incluso si el enfoque ha sido el mismo, se han usado diferentes metodologías en relación a las medidas, puntos de referencia o marcadores moleculares, así como diferentes procedimientos estadísticos. Por lo tanto, existe la necesidad de establecer métodos comunes en todos los países para tener resultados que puedan ser directamente comparados. Este es uno de los objetivos del GT4 del proyecto COLOSS.