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Anesthetization of bees for identification

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  • Dennis E Johnson
    All, I sent Sam information about a method that I have been using in the field to anesthetize bees for identification, especially Bombus sp. in the spring when
    Message 1 of 3 , Dec 6, 2013
    All,

    I sent Sam information about a method that I have been using in the field to anesthetize bees for identification, especially Bombus sp. in the spring when most will be queens and he suggested that I send it to the group.

    Below is a picture of the apparatus I use. It is a Bike Planet tire inflater (< $20.00) that uses 16 g threaded cartridges and can be adjusted for flow manually and shut off. It seems to leak out after about a day if you don't use it all and cartridges can be gotten for $1.00 each if you shop around. To use, I capture the bee in a small container like a pill bottle that I have drilled a small hole in to be able to insert a ball inflator needle screwed into the inflator. Ahead of the insertion, I adjust the flow for a small amount and then insert the needle into the container. It takes only about 2 - 3 seconds for the bee to pass out but I usually let them lie in the CO2 atmosphere for a minute or so after I shut off the flow and it takes about 5 minutes or more for them to wake up and eventually fly off. You have to experiment with this some for an optimum "out" time.
    I'd say that they don't curl up…see photos of the pocket sized apparatus and bees that I CO2'd. They will occasionally move a leg or antenna while they are out, but not a problem for single shot photos…..different issue for stacking.

    I have not had a bee that did not recover and fly off even if I left them in the CO2 for a couple of minutes, however, I am not sure what it may do to their memory and other functions. This spring I may try to find a new queen in her new nest and perform the procedure on her to see if she raises a viable colony afterwards.

    Since CO2 is heavier than air, I think that they could be immersed in the CO2 in an open dish for photos…..have not tried yet. Also I have chilled them to extend the "out" time, but they tend to curl some. I get about 7 - 8 injections per cartridge unless I over shoot the flow rate. If you were doing a lot of bees, you could get paint ball CO2 cylinders or beer tap cylinders which could be regulated better and shut off completely, but would be less portable, however, good in the lab.
    I f you have any questions, let me know.

    Denny Johnson, retired engineer doing dragonfly and bee research as a volunteer
  • David Inouye
    Some papers about effects of CO2. David Beckmann, H. (1974). Beeinflussung des Gedder Honigbiene durch Narkose, Kund Stress. Journal of Comparative
    Message 2 of 3 , Dec 6, 2013
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      Some papers about effects of CO2.

      David

      Beckmann, H. (1974). "Beeinflussung des Gedder Honigbiene durch Narkose, Kund Stress." Journal of Comparative Physiology 94: 249-266.
               Retrograde amnesia occurs if, shortly after training, bees are exposed to supercooling, CO2 narcosis, or to a stress situation caused by captivity. Memory loss was about 50%, and memory was not recovered within 24 hours.

      Ebadi, R., et al. (1980). "Effects of carbon dioxide and low temperature narcosis on honey bees, Apis mellifera." Environmental Entomology 9(1): 144-147.
               Short exposure of < 2 min to pure CO2 or 3 min to -20C did not alter the orientation or return of released foragers to colonies. CO2 treatments of 30, 60, and 120 sec reduced survival and pollen-gathering behavior.

      Hoover, S. E. R., et al. (2012). "Warming, CO2, and nitrogen deposition interactively affect a plant-pollinator mutualism." Ecology Letters 15(3): 227-234.
               Environmental changes threaten plant-pollinator mutualisms and their critical ecosystem service. Drivers such as land use, invasions and climate change can affect pollinator diversity or species encounter rates. However, nitrogen deposition, climate warming and CO2 enrichment could interact to disrupt this crucial mutualism by altering plant chemistry in ways that alter floral attractiveness or even nutritional rewards for pollinators. Using a pumpkin model system, we show that these drivers non-additively affect flower morphology, phenology, flower sex ratios and nectar chemistry (sugar and amino acids), thereby altering the attractiveness of nectar to bumble bee pollinators and reducing worker longevity. Alarmingly, bees were attracted to, and consumed more, nectar from a treatment that reduced their survival by 22%. Thus, three of the five major drivers of global environmental change have previously unknown interactive effects on plant-pollinator mutualisms that could not be predicted from studies of individual drivers in isolation.

      Kukuk, P. F., et al. (1997). "Larval ejection behavior in Bombus occidentalis in response to CO
      2 - or N2 -induced narcosis." Journal of the Kansas Entomological Society 70(4): 359-361.

               Previous studies have suggested that CO2 narcosis results in larval ejection by bumble bees. An experiment is described demonstrating that both CO2- and N2-induced narcosis result in chewing of the waxen larval envelope, a distinctive behavior that precedes larval ejection, in the bumble bee, Bombus occidentalis. These observations suggest that a lack of O2 rather than an excess of CO2 may be responsible for the behavioral changes following CO2 narcosis. Although the proximate and ultimate factors leading to larval ejection are not well understood, they could possibly be related to regulation of respiratory gases in the underground burrows of Bombus occidentalis.

              
      Shalimov, I. I., et al. (1994). "Influence of carbon-dioxide narcotization on bumble bees behaviour." Vestnik Zoologii 28(3): 82-84.
               Narcotization of a bumble bee family with carbon dioxide results in decrease of its foraging activity and increase of food demand, causing energetic balance shift to the negative side.  As a result of starvation, a part of brood is ejected from the nest.

    • Weber, Don
      Thank you David for this nice set of citations. I wish that those working with ALL insects using CO2, in particular with undefined durations and
      Message 3 of 3 , Dec 6, 2013
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        Thank you David for this nice set of citations.  I wish that those working with ALL insects using CO2, in particular with undefined durations and concentrations, would be cognizant of the potentially radical effects, especially for behavioral studies.

        The title of the first one puzzled me; The journal citation is correct, but the title should be:
        "Beeinflussung des Gedächtnisses der Honigbiene durch Narkose, Kühlung und Streß."
        = Impact on memories in the honeybee through narcosis, cooling, and stress


        ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~

        Donald C. Weber, Research Entomologist & Lead Scientist

        USDA Agricultural Research Service

        Invasive Insect Biocontrol & Behavior Laboratory

        Bldg. 011A, Rm. 107, BARC-West Beltsville, MD 20705  USA

        Don.Weber@...



        From: beemonitoring@yahoogroups.com [beemonitoring@yahoogroups.com] on behalf of David Inouye [inouye@...]
        Sent: Friday, December 06, 2013 12:34 PM
        To: Bee monitoring Group
        Subject: Re: [beemonitoring] Anesthetization of bees for identification

         

        Some papers about effects of CO2.

        David

        Beckmann, H. (1974). "Beeinflussung des Gedder Honigbiene durch Narkose, Kund Stress." Journal of Comparative Physiology 94: 249-266.

                 Retrograde amnesia occurs if, shortly after training, bees are exposed to supercooling, CO2 narcosis, or to a stress situation caused by captivity. Memory loss was about 50%, and memory was not recovered within 24 hours.

        Ebadi, R., et al. (1980). "Effects of carbon dioxide and low temperature narcosis on honey bees, Apis mellifera." Environmental Entomology 9(1): 144-147.
                Short exposure of < 2 min to pure CO2 or 3 min to -20C did not alter the orientation or return of released foragers to colonies. CO2 treatments of 30, 60, and 120 sec reduced survival and pollen-gathering behavior.

        Hoover, S. E. R., et al. (2012). "Warming, CO2, and nitrogen deposition interactively affect a plant-pollinator mutualism." Ecology Letters 15(3): 227-234.
                 Environmental changes threaten plant-pollinator mutualisms and their critical ecosystem service. Drivers such as land use, invasions and climate change can affect pollinator diversity or species encounter rates. However, nitrogen deposition, climate warming and CO2 enrichment could interact to disrupt this crucial mutualism by altering plant chemistry in ways that alter floral attractiveness or even nutritional rewards for pollinators. Using a pumpkin model system, we show that these drivers non-additively affect flower morphology, phenology, flower sex ratios and nectar chemistry (sugar and amino acids), thereby altering the attractiveness of nectar to bumble bee pollinators and reducing worker longevity. Alarmingly, bees were attracted to, and consumed more, nectar from a treatment that reduced their survival by 22%. Thus, three of the five major drivers of global environmental change have previously unknown interactive effects on plant-pollinator mutualisms that could not be predicted from studies of individual drivers in isolation.

        Kukuk, P. F., et al. (1997). "Larval ejection behavior in Bombus occidentalis in response to CO
        2 - or N2 -induced narcosis." Journal of the Kansas Entomological Society 70(4): 359-361.

                 Previous studies have suggested that CO2 narcosis results in larval ejection by bumble bees. An experiment is described demonstrating that both CO2- and N2-induced narcosis result in chewing of the waxen larval envelope, a distinctive behavior that precedes larval ejection, in the bumble bee, Bombus occidentalis. These observations suggest that a lack of O2 rather than an excess of CO2 may be responsible for the behavioral changes following CO2 narcosis. Although the proximate and ultimate factors leading to larval ejection are not well understood, they could possibly be related to regulation of respiratory gases in the underground burrows of Bombus occidentalis.

                
        Shalimov, I. I., et al. (1994). "Influence of carbon-dioxide narcotization on bumble bees behaviour." Vestnik Zoologii 28(3): 82-84.
                 Narcotization of a bumble bee family with carbon dioxide results in decrease of its foraging activity and increase of food demand, causing energetic balance shift to the negative side.  As a result of starvation, a part of brood is ejected from the nest.





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