Compliance with recommended Varroa destructor treatment regimens improves the survival of honey bee colonies over winter.

The ectoparasitic mite Varroa destructor affects honey bee colony health and survival negatively, thus compelling beekeepers to treat their colonies every year. A broadly used mite control regimen is based on two organic molecules: formic and oxalic acids. To ensure optimal efficiency, several applications of these acids at pre-defined time points are recommended. These recommendations are mainly based on experiments conducted under controlled conditions. Studies evaluating the effectiveness under natural field conditions are lacking. We enrolled 30 beekeepers in a longitudinal study in three cantons in Switzerland and monitored the management and health of their colonies for two years. We assessed compliance with mite control recommendations and measured V. destructor infestation rates, indexes of colony productivity (brood size and honey harvest), and colony mortality in 300 colonies. We observed a 10-fold increased risk of colony death when beekeepers deviated slightly from the recommended treatment regimen compared to compliant beekeepers (odds ratio: 11.9, 95% CI: 2.6-55.2, p = 0.002). The risk of colony death increased 25-fold in apiaries with substantial deviations from the recommendations (odds ratio: 50.4, 95% CI: 9.7-262.5, p < 0.0001). The deviations led to increased levels of V. destructor infestation ahead of wintering, which was likely responsible for colony mortality. After communicating the apparent link between low compliance and poor colony survival at the end of the first year to the beekeepers, we observed better compliance and colony survival in the second year. Our results highlight the positive impact of compliance with the recommended V. destructor treatment regimen on the health of honeybee colonies and the need to better communicate the consequences of deviating from the recommendations to improve compliance. Compliance also occasionally decreased, which hints at concept implementation constraints that could be identified and possibly addressed in detail with the help of social sciences to further promote honey bee health.

Land conversion and pesticide use degrade forage areas for honey bees in America's beekeeping epicenter.

A diverse range of threats have been associated with managed-bee declines globally. Recent increases of two known threats, land-use change and pesticide use, have resulted from agricultural expansion and intensification notably in the top honey-producing state in the United States: North Dakota. This study investigated the dual threat from land conversion and pesticide use surrounding ~14,000 registered apiaries in North Dakota from 2001 to 2014. We estimated the annual total insecticide use (kg) on major crops within 1.6 km of apiary sites. Of the eight insecticides quantified, six showed significant increasing trends over the time period. Specifically, applications of the newly established neonicotinoids Chlothianidin, Imidacloprid and Thiamethoxam, increased annually by 1329 kg, 686 kg, 795 kg, respectively. Also, the use of Chlorpyrifos, which was well-established in the state by 2001 and is highly toxic to honey bees, increased by ~8,800 kg annually from 6,500 kg in 2001 to 115,000 kg in 2014 on corn, soybeans and wheat. We further evaluated the relative quality changes of natural/semi-natural land covers surrounding apiaries in 2006, 2010 and 2014, a period of significant increases in cropland area. In areas surrounding apiaries, we observed changes in multiple indices of forage quality that reflect the deteriorating landscape surrounding registered apiary sites due to land-use change and pesticide-use increases. Overall, our results suggest that the application of foliar-applied insecticides, including pyrethroids and one organophosphate, increased surrounding apiaries when the use of neonicotinoid seed treatments surged and the area for producing corn and soybeans expanded. Spatially, these threats were most pronounced in southeastern North Dakota, a region hosting a high density of apiary sites that has recently experienced corn and soybean expansion. Our results highlight the value of natural and semi-natural land covers as sources of pollinator forage and refugia for bees against pesticide exposure. Our study provides insights for targeting conservation efforts to improve forage quality benefiting managed pollinators.

Dyeing but not dying: Colourful dyes as a non-lethal method of food labelling for in vitro-reared honey bee (Apis mellifera) larvae.

Several environmental factors (e.g. food source, pesticides, toxins, parasites and pathogens) influence development and maturation of honey bees (Apis mellifera). Therefore, controlled experimental conditions are mandatory when studying the impact of environmental factors: particularly food quality and nutrient consumption. In vitro larval rearing is a standard approach for monitoring food intake of larvae and the labelling of food is necessary to quantify intake in controlled feeding experiments. Here, we tested the suitability of two food dyes, Allura Red and Brilliant Blue, in an experimental set up using in vitro reared honey bee larvae and freshly hatched adult workers. Absorbance of both dyes was measured, in food and dye-fed larvae, to determine the optimal dye concentrations for accurate detection and quantification. By quantifying relative dye concentrations in dye mixtures, relative concentrations of mixed dyes can be estimated independent of the total food consumed by the larvae. Survival assays were conducted to test the impact of both dyes on larval and worker bee survival. Worker bees showed no increase in adult mortality, when fed with dyed honey. Larval survival was not significantly different until the late pupal stage. The physiological impact of dye feeding was tested by measuring larval immune response. No changes in innate immune gene expression were detectable for larvae fed with dyed and non-dyed food. In conclusion, we established a non-invasive food labelling protocol for food intake quantification in in vitro reared honey bee larvae, using non-toxic, inexpensive, and easy to apply food dyes.

Parasaccharibacter apium, gen. nov., sp. nov., Improves Honey Bee (Hymenoptera: Apidae) Resistance to Nosema.

The honey bee, Apis mellifera L., is host to a variety of microorganisms. The bacterial community that occupies the adult worker gut contains a core group of approximately seven taxa, while the hive environment contains its own distribution of bacteria that is in many ways distinct from the gut. Parasaccharibacter apium, gen. nov., sp. nov., is a hive bacterium found in food stores and in larvae, worker jelly, worker hypopharyngeal glands, and queens. Parasaccharibacter apium increases larval survival under laboratory conditions. To determine if this benefit is extended to colonies in the field, we tested if P. apium 1) survives and reproduces in supplemental pollen patty, 2) is distributed throughout the hive when added to pollen patty, 3) benefits colony health, and 4) increases the ability of bees to resist Nosema. Parasaccharibacter apium survived in supplemental diet and was readily consumed by bees. It was distributed throughout the hive under field conditions, moving from the pollen patty to hive larvae. While P. apium did not significantly increase colony brood production, food stores, or foraging rates, it did increase resistance to Nosema infection. Our data suggest that P. apium may positively impact honey bee health.

Foreign matter contaminating ethanolic extract of propolis: a filth-test survey comparing products from small beekeeping farms and industrial producers.

Propolis is a resinous material collected by honeybees from the exudates and buds of plants. It has been widely used as a remedy by humans since ancient times, as well as for dietary supplements and cosmetics. European legislation recently focused on the quality and hygiene standards of foods, including beehive products, and extensive efforts have been made to avoid the presence of chemical contaminants, whilst in contrast few studies have investigated the magnitude of contamination by physical ones. We conducted a filth-test survey to evaluate the contamination of ethanolic extract of propolis by foreign materials. We also compared the abundance of contaminants in propolis extracts currently marketed by small beekeepers and industrial producers. We found different foreign materials in the ethanol extract of propolis. Contaminants differed in abundance, with a higher number of carbon particles (small beekeepers: 2.70 ± 0.63; industrial producers: 1.25 ± 0.49; mean (n/30 ml) ± SE) and other inorganic fragments (small beekeepers: 3.50 ± 0.31; industrial producers: 3.88 ± 1.11) than arthropod fragments (small beekeepers: 0.30 ± 0.21; industrial producers: 0.38 ± 0.26) and mammal hairs (small beekeepers: 0.10 ± 0.10; industrial producers: 0.38 ± 0.26). No differences in the abundance of foreign matter between propolis from small beekeepers and industrial producers were found, allowing us to point out an increased awareness by small producers of issues inherent in hygiene management. Contamination of propolis extracts by animal body parts, such as insect fragments, mites and rodent hairs, indicates poor management of hygiene in the production process and low effectiveness of the filtration phase. Animal-borne contaminants can act as pathogen vectors as well as introducing dangerous allergens when ingested or applied to human skin. The filth-test applied to ethanolic propolis extract quality control can be considered a promising tool, also for small beekeeper activities, since it is cheap and allows the quick interpretation of results.

Holistic screening of collapsing honey bee colonies in Spain: a case study.

BACKGROUND: Here we present a holistic screening of collapsing colonies from three professional apiaries in Spain. Colonies with typical honey bee depopulation symptoms were selected for multiple possible factors to reveal the causes of collapse. RESULTS: Omnipresent were Nosema ceranae and Lake Sinai Virus. Moderate prevalences were found for Black Queen Cell Virus and trypanosomatids, whereas Deformed Wing Virus, Aphid Lethal Paralysis Virus strain Brookings and neogregarines were rarely detected. Other viruses, Nosema apis, Acarapis woodi and Varroa destructor were not detected. Palinologic study of pollen demonstrated that all colonies were foraging on wild vegetation. Consequently, the pesticide residue analysis was negative for neonicotinoids. The genetic analysis of trypanosomatids GAPDH gene, showed that there is a large genetic distance between Crithidia mellificae ATCC30254, an authenticated cell strain since 1974, and the rest of the presumed C. mellificae sequences obtained in our study or published. This means that the latter group corresponds to a highly differentiated taxon that should be renamed accordingly. CONCLUSION: The results of this study demonstrate that the drivers of colony collapse may differ between geographic regions with different environmental conditions, or with different beekeeping and agricultural practices. The role of other pathogens in colony collapse has to bee studied in future, especially trypanosomatids and neogregarines. Beside their pathological effect on honey bees, classification and taxonomy of these protozoan parasites should also be clarified.

Prospects of apicultural entrepreneurship in coastal districts of eastern India: a melissopalynological evaluation.

A melissopalynological analysis of fifty-one natural honey samples (twenty four spring, fifteen summer and twelve winter) collected during 2010-2011 from two east-coastal districts (20(0)20/ to 22(0)11/ N, 82(0)39/ to 87(0)01/ E) of Orissa, India was performed. Out of 37 unifloral samples found 25 were contributed by Apis cerana indica, seven by A. dorsata and the remaining five by A. florea. Out of 14 multifloral samples five were contributed by A. cerana indica, five by A. dorsata and the remaining four by A. florea. Principal component analysis confirmed the palynological classification of the unifloral honey samples. Eighty-two bee-plant taxa belonging to forty four families were recovered. The predominant nectariferous taxa of the spring season were Acanthus ilicifolius, Avicennia marina, Bruguiera gymnorrhiza, Cocos nucifera, Eucalyptus globulus, Phoenix paludosa, Pongamia pinnata, Prosopis juliflora, Sonneratia apetala and Syzygium cumini. In the summer the predominant nectariferous taxa were Borassus flabellifer, C. nucifera, E. globulus, Syzygium cumini, Terminalia arjuna, Aegiceras corniculatum, P. paludosa and Sonneratia apetala while those of the winter were Brassica nigra, Coriandrum sativum, Zizyphus jujuba, Alstonia scholaris, E. globulus and Bruguiera gymnorrhiza. Very low (<0.09) HDE/P for 98% of the samples and absence of toxic palynotaxa assure that these honeys are suitable for human consumption. Quite extended honey flow period with spring and summer as best forage seasons for the honeybees and occurrence of 82% of these honeys with APC Group II, III and IV justify the sustainability of the present study area for establishing moderate to large-scale apicultural entrepreneurship. This should improve the socio-economic status of the people of this region.

Survey of Italian honeys for the presence of foreign matter using the filth test.

Honey is an ancient food that has always been considered a natural and healthy product, free of contaminants. However, it can contain toxic substances, such as antibiotics, pesticides and heavy metals, as well as foreign matter (e.g. arthropod body parts and microbial contaminants), working as allergens and vectors of human pathogens. In this study we used the filth test to evaluate the abundance of foreign matter in 70 Italian honeys, including Castanea sativa, Robinia pseudoacacia and multi-floral honeys, the latter both from small beekeeping farms and industrial producers. The abundance of different foreign matter varied in honeys, with a higher number of carbon particles and other inorganic fragments, followed by fragments of animal origin. This latter included insects (Diptera Brachycera larvae and Strepsiptera), their cuticular fragments (mainly Coleoptera, Hymenoptera and Lepidoptera), mites (mainly Glycyphagidae, Acaridae and Tarsonemidae) and mammal hairs. No differences were recorded in the abundance of foreign matter among different kinds of honey, as well as between honeys from small and large-sized producers. Foreign matter found in honey provided functional information to evaluate honey quality standards in apiary, honey extraction and packaging phases. Overall, the filth test method applied to honey quality control can be considered an excellent tool, also for small beekeeping farms, since it allows rapid and frequent quality checking of the production process. This method is cheap, requires minimal instrumental equipment and results can be interpreted quickly.

Evaluación de dos sistemas de secador y dos tiempos de secado en las características microbiológicas, físico-químicas y sensoriales del polen de abejas / Evaluation of two dryer systems and two drying times in the microbiological, physical-chemical and sensory characteristics of bee pollen

El polen de abejas es un alimento natural que actualmente es utilizado en la medicina tradicional, en industrias farmacéuticas e industria alimentaria. La calidad microbiológica y sensorial del polen depende del tiempo y del tipo de secador utilizado durante el procesamiento. El objetivo de este estudio fue determinar el efecto de dos tipos de secador y dos tiempos de secado en las características microbiológicas, físico-químicas y sensoriales del polen de abeja. El diseño experimental utilizado fue un Diseño Completamente al Azar (DCA) con un arreglo factorial de 2x2, evaluando el tipo secador (solar y horno) y el tiempo de secado (1 y 3 horas) más un control. Microbiológicamente se hicieron recuentos de coliformes totales, aerobios mesófilos, hongos y levaduras. Se realizaron análisis químicos (humedad y aw), físicos (color e impurezas) y sensoriales (apariencia, color y aceptación general). El polen secado en horno, presentó menores recuentos de aerobios mesófilos independiente del tiempo de secado. La carga de hongos y levaduras disminuyó luego del secado independiente del tiempo y tipo de secador. Todos los tratamientos presentaron ausencia de coliformes totales. En las propiedades físicas y sensoriales los tratamientos no presentaron diferencias significativas (P>0.05). Se recomienda aumentar el tiempo de secado a seis horas.

Fast determination of royal jelly freshness by a chromogenic reaction.

UNLABELLED: Royal jelly is one of the most important products of honeybees. Given its role in development of bee brood into fertile individuals of the royal caste it is also used in health products for human consumption. Royal jelly spoils and loses its health-promoting properties depending on storage duration and conditions. To ensure product quality before selling, it is therefore necessary to assess royal jelly freshness. Many indexes of freshness have been suggested, but they all lack reliability or require complex and time-consuming analyses. Here we describe a method to detect royal jelly freshness based on a chromogenic reaction between royal jelly and HCl. We demonstrate that analyses based on color parameters allow for the discrimination of royal jelly samples based on the duration of their storage. Color parameters of royal jelly stored at -18 and 4 °C for 28 d remained comparable to that of fresh samples, which supports the reliability of the method. The method of freshness determination described is practical, cheap, and fast and can thus be used in real-time when trading royal jelly. PRACTICAL APPLICATION: The method developed can be used to assess royal jelly freshness. It is practical, cheap, and fast and can thus be used in real-time when trading royal jelly.

Treatment with synthetic brood pheromone (SuperBoost) enhances honey production and improves overwintering survival of package honey bee (Hymenoptera: Apidae) colonies.

We evaluated a year-long treatment regime testing synthetic, 10-component, honey bee, Apis mellifera L. (Hymenoptera: Apidae), brood pheromone (SuperBoost; Contech Enterprises Inc., Delta, BC, Canada) on the productivity and vigor of package bee colonies in the lower Fraser Valley of British Columbia, Canada. Fifty-eight newlyestablished 1.3-kg (3-lb) colonies treated three times with SuperBoost at 5-wk intervals starting 30 April 2009 were compared with 52 untreated control colonies. Treated colonies produced 84.3% more honey than untreated control colonies. By 8 September 2009, SuperBoost-treated colonies had 35.4% more adults than untreated colonies. By 28 September, net survival of treated and control colonies was 72.4 and 67.3%, respectively. On 5 October, treated and control colonies were divided into two additional groups, making up four cohorts: SuperBoost-treated colonies treated again during fall and spring build-up feeding with pollen substitute diet (BeePro, Mann Lake Ltd., Hackensack, MN; TIT); controls that remained untreated throughout the year (CCC); colonies treated with SuperBoost in spring-summer 2009 but not treated thereafter (TCC); and original control colonies treated with SuperBoost during the fall and spring build-up feeding periods (CTT). There was no difference among cohorts in consumption of BeePro during fall feeding, but TTT colonies (including daughter colonies split off from parent colonies) consumed 50.8% more diet than CCC colonies during spring build-up feeding. By 21 April, the normalized percentages of the original number of colonies remaining (dead colonies partially offset by splits) were as follows: CCC, 31.4%; CTT, 43.8%; TCC, 53.59%; and TTT, 80.0%. The net benefit of placing 100 newly established package bee colonies on a year-long six-treatment regime with SuperBoost would be US$6,202 (US$62.02 per colony). We conclude that treatment with SuperBoost enhanced the productivity and survival of package bee colonies and hypothesize that similar results could be achieved with established colonies.

High levels of miticides and agrochemicals in North American apiaries: implications for honey bee health.

BACKGROUND: Recent declines in honey bees for crop pollination threaten fruit, nut, vegetable and seed production in the United States. A broad survey of pesticide residues was conducted on samples from migratory and other beekeepers across 23 states, one Canadian province and several agricultural cropping systems during the 2007-08 growing seasons. METHODOLOGY/PRINCIPAL FINDINGS: We have used LC/MS-MS and GC/MS to analyze bees and hive matrices for pesticide residues utilizing a modified QuEChERS method. We have found 121 different pesticides and metabolites within 887 wax, pollen, bee and associated hive samples. Almost 60% of the 259 wax and 350 pollen samples contained at least one systemic pesticide, and over 47% had both in-hive acaricides fluvalinate and coumaphos, and chlorothalonil, a widely-used fungicide. In bee pollen were found chlorothalonil at levels up to 99 ppm and the insecticides aldicarb, carbaryl, chlorpyrifos and imidacloprid, fungicides boscalid, captan and myclobutanil, and herbicide pendimethalin at 1 ppm levels. Almost all comb and foundation wax samples (98%) were contaminated with up to 204 and 94 ppm, respectively, of fluvalinate and coumaphos, and lower amounts of amitraz degradates and chlorothalonil, with an average of 6 pesticide detections per sample and a high of 39. There were fewer pesticides found in adults and brood except for those linked with bee kills by permethrin (20 ppm) and fipronil (3.1 ppm). CONCLUSIONS/SIGNIFICANCE: The 98 pesticides and metabolites detected in mixtures up to 214 ppm in bee pollen alone represents a remarkably high level for toxicants in the brood and adult food of this primary pollinator. This represents over half of the maximum individual pesticide incidences ever reported for apiaries. While exposure to many of these neurotoxicants elicits acute and sublethal reductions in honey bee fitness, the effects of these materials in combinations and their direct association with CCD or declining bee health remains to be determined.