Standard methods for<i>Apis mellifera</i>anatomy and dissection

Carreck, Norman; Andree, Michael; Brent, Colin S.; Cox-Foster, Diana; Dade, H. A.; Ellis, Jamie; Hatjina, Fani; Englesdorp, Dennis van

doi:10.3896/ibra.1.52.4.03

Cited by 128 publications

(102 citation statements)

References 24 publications

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“…To evaluate HPG development, on days 8 and 12 of the experiment, 21 bees per treatment per day (7 bees × 3 genetic lineages) were sacrificed for HPG dissection, according to previously published protocols . Briefly, the bees were anesthetized with CO 2 , decapitated, and the heads were embedded in a beeswax dissection plate.…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, the bees were anesthetized with CO 2 , decapitated, and the heads were embedded in a beeswax dissection plate. Next, the heads were dissected in insect saline (1 L distilled water with 7.5 g NaCl, 2.38 g Na 2 HPO 4 and 2.72 g KH 2 PO 4 ) using an Olympus SZ61 stereomicroscope (Olympus, Tokyo, Japan). The isolated glands were stained with ∼250 μL of 0.25% (w/v) Coomassie Brilliant Blue R‐250 for ∼3 s and then stored in insect saline at 4°C for 24–48 h until digital image capture was performed.…”

Section: Methodsmentioning

confidence: 99%

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Effects of chronic dietary thiamethoxam and prothioconazole exposure onApis melliferaworker adults and brood

Wood

Mattos

Kozii

et al. 2019

Pest Management Science

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BACKGROUND Chronic exposure of honey bees (Apis mellifera Linnaeus) to the neonicotinoid thiamethoxam and the fungicide prothioconazole is common during foraging in agricultural landscapes. We evaluated the survival and hypopharyngeal gland development of adult worker honey bees, and the survival of the worker brood when chronically exposed to thiamethoxam or thiamethoxam and prothioconazole in combination. RESULTS We found that 30 days of exposure to 40 μg kg–1 of thiamethoxam significantly (P < 0.001) increased the frequency of death in worker adults by four times relative to solvent control. The worker brood required 23 times higher doses of thiamethoxam (1 mg L–1 or 909 μg kg–1) before a significant (P = 0.04), 3.9 times increase in frequency of death was observed relative to solvent control. No additive effects of simultaneous exposure of worker adults or brood to thiamethoxam and prothioconazole were observed. At day 8 and day 12, the hypopharyngeal gland acinar diameter was not significantly different (P > 0.05) between controls and adult workers exposed to thiamethoxam and/or prothioconazole. CONCLUSION These results indicate that chronic exposure to field‐realistic doses of thiamethoxam and/or prothioconazole are unlikely to affect the survival of adult workers and brood. © 2019 Society of Chemical Industry

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Effects of chronic dietary thiamethoxam and prothioconazole exposure onApis melliferaworker adults and brood

Wood

Mattos

Kozii

et al. 2019

Pest Management Science

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“…About 200 returning honey bee foragers per sample were caught at the hive entrances by using a vacuum collector and immediately transferred to dry ice and stored frozen (between −10 °C and −43 °C) until dissection. The honey stomachs of the collected honey bees were dissected in the laboratory (according to Carreck et al 2013) and the nectar (minimum of 200 mg) was discharged into a 15 ml tube. All nectar samples were stored frozen (between −10 °C and −43 °C) until analysis.…”

Section: Methodsmentioning

confidence: 99%

Large-scale monitoring of effects of clothianidin-dressed oilseed rape seeds on pollinating insects in northern Germany: residues of clothianidin in pollen, nectar and honey

Rolke

Persigehl²,

Peters³

et al. 2016

Ecotoxicology

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This study was part of a large-scale monitoring project to assess the possible effects of Elado® (10 g clothianidin & 2 g β-cyfluthrin/kg seed)-dressed oilseed rape seeds on different pollinators in Northern Germany. Firstly, residues of clothianidin and its active metabolites thiazolylnitroguanidine and thiazolylmethylurea were measured in nectar and pollen from Elado®-dressed (test site, T) and undressed (reference site, R) oilseed rape collected by honey bees confined within tunnel tents. Clothianidin and its metabolites could not be detected or quantified in samples from R fields. Clothianidin concentrations in samples from T fields were 1.3 ± 0.9 μg/kg and 1.7 ± 0.9 μg/kg in nectar and pollen, respectively. Secondly, pollen and nectar for residue analyses were sampled from free flying honey bees, bumble bees and mason bees, placed at six study locations each in the R and T sites at the start of oilseed rape flowering. Honey samples were analysed from all honey bee colonies at the end of oilseed rape flowering. Neither clothianidin nor its metabolites were detectable or quantifiable in R site samples. Clothianidin concentrations in samples from the T site were below the limit of quantification (LOQ, 1.0 µg/kg) in most pollen and nectar samples collected by bees and 1.4 ± 0.5 µg/kg in honey taken from honey bee colonies. In summary, the study provides reliable semi-field and field data of clothianidin residues in nectar and pollen collected by different bee species in oilseed rape fields under common agricultural conditions.

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“…The drones in the second group, referred to as SC, were individually treated with chloroform for the partial eversion of the endophallus and then their abdomens were squeezed the same way as the drones in the SM group for full eversion and ejaculation (Mackensen and Ruttner, 1976). The drones allocated to the third group, referred to as S, after falling into a chill-coma, as a result of keeping them for 15 -20 minutes at a temperature of 6°C, were dissected and their reproductive organs were assessed (Dade, 1994;Carreck et al, 2013 experiment was repeated twice. In the first replication, 133 drones (45 were dissected, 40 in group MS, and 48 in group SC) were examined.…”

Section: Methodsmentioning

confidence: 99%

The Ability of Honey Bee Drones to Ejaculate

Czekońska

Chuda-Mickiewicz

2015

Journal of Apicultural Science

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A b s t r a c t The effectiveness of two methods of collecting semen from honeybee Apis mellifera drones was compared, and the reasons for problems with ejaculating semen were analysed. Among 275 drones, 100 were stimulated to release semen using a manual method, 100 with the use of chloroform, and from 75 drones the reproductive organs were dissected for analysis and evaluation. It was found that the principal causes of problems that drones had with ejaculating their semen were anatomical changes or a delay in the development of the mucus glands. It was also found that the method employing chloroform was less efficient in the first phase of eversion of the endophallus, compared with the manual method. The method with the use of chloroform allows the determination of the proportion of drones, which do not evert the endophallus because of poor or delayed development of mucus glands, as well as the proportion of drones which evert the organ, but do not ejaculate semen because of the absence of semen in the seminal vesicles.Keywords: Apis mellifera, drone, eversion, reproductive organ. INTRODUCTIONIn a honeybee colony drones are responsible for producing semen and transmitting it to the queen during copulation. In drones, the spermatozoa are produced in the pre-imago stage of development. In the first days of a drone's life the spermatozoa are moved from the testes to the seminal vesicles, where they are stored until mating. Immediately after emerging from the cells, drones are not able to copulate. They reach the ability to mate in 9 -12 days after emergence (Bishop, 1920;Ruttner and Tryasko, 1976). Drones mate with the queen at the age of 15 -23 days, the average being 21 days (Couvillon et al., 2010). Because of the limited number of young queens, only a few drones have the chance to pass on their semen during copulation. The majority of drones die because of age, diseases, or predators (Free and Williams, 1975;Fukuda and Ohtani, 1977;Rueppell et al., 2005;Boes, 2010). A drone does not survive copulation (Witherell, 1965). On average, the drones that do not have the chance to copulate with a queen live 30 days (Rueppell et al., 2005). Colonies differ in terms of the number of drones reared and their reproductive activity (Kraus et al., 2003;Boes, 2010). The reproductive success of drones depends much on the size of the colony and the conditions prevailing during rearing and maturing (Jaycox, 1961;Boes, 2010;Bieńkowska et al., 2011;Mazeed, 2011;Abdelkader et al.;. The drones reared in larger colonies achieve greater reproductive success; more often copulate with the queen, and also have a larger share in the number of offspring (Kraus et al., 2003). The drones reared under optimum thermal and feeding conditions have greater chance of mating (Jaycox, 1961;Rueppell et al., 2006;Czekońska et al., 2013). Drones reared in colonies of different quality, differ in the age when they reach the ability to copulate with the queen (Rhodes et al., 2011;Czekońska et al., 2015). Therefore, in practice, the age of drones is not...

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Standard methods forApis melliferaanatomy and dissection

Cited by 128 publications

References 24 publications

Effects of chronic dietary thiamethoxam and prothioconazole exposure onApis melliferaworker adults and brood

Effects of chronic dietary thiamethoxam and prothioconazole exposure onApis melliferaworker adults and brood

Large-scale monitoring of effects of clothianidin-dressed oilseed rape seeds on pollinating insects in northern Germany: residues of clothianidin in pollen, nectar and honey

The Ability of Honey Bee Drones to Ejaculate

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