Duchateau and Velthuis ovarian classification (Halictidae)

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Method synopsis

Collection of female halictid bees at nest sites using nets or vacuum devices, with individual storage and cryopreservation for chemical analysis. Includes morphological measurement and ovarian stage classification for caste determination.

Synthesized from method descriptions across 5 papers using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Chemical Variation among Castes, Female Life Stages and Populations of the Facultative Eusocial Sweat Bee <i> Halictus rubicundus </i> (Hymenoptera: Halictidae) (2021), Journal of Chemical Ecology. Implementations in other papers (listed below) may differ.

1. Bee collection from nest sites
   Collected Halictus rubicundus females at their nest sites using an insect net or a vacuum suction device. Specimens were stored individually in small plastic Eppendorf tubes (1.5 ml).
   Quantities: Total of 158 females: 20 solitary nest foundresses, 20 solitary breeding females from Belfast; 21 nest foundresses, 22 social breeding queens, 47 workers from European social populations; 14 solitary breeding females from Gothic, Colorado; 37 breeding queens and workers from Logan, Utah social populationDuration: Collections made during spring and summer 2011 in Belfast, spring and summer 2013 in Bonn and Hayingen, summer 2016 in Gothic, summer 2016 in LoganConditions: Collections made at nest sites during active periods
   Equipment: insect net, vacuum suction device, plastic Eppendorf tubes (1.5 ml)
2. Specimen preservation
   Killed bees by freezing at -40°C for further use in chemical analyses. Chemical compounds, especially cuticular hydrocarbons, are known to exhibit a long-term stability.
   Quantities: All collected specimens frozen individuallyDuration: Preserved for chemical analysis (collection years varied: 2011-2016)Conditions: Storage temperature of -40°C
   Equipment: freezer capable of -40°C
3. Morphological measurements
   After extraction of cuticle surface, all bees were dissected under a stereomicroscope to check ovarian stage, which was classified into five categories according to Duchateau and Velthuis (1989) for physiological state and caste determination. Head width (maximum width of the head including the compound eyes) was used as a proxy of size and was measured for each female using an eyepiece graticule.

Typical Equipment

• insect net
• vacuum suction device
• plastic Eppendorf tubes (1.5 ml)
• freezer capable of -40°C
• stereomicroscope
• eyepiece graticule

Output Measurements

• preserved bee specimens
• head width measurements
• ovarian stage classifications
• caste assignments

Papers Using This Protocol (4)

Species Studied With This Protocol

Places Where Used