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A Proboscis Extension Response Protocol for Investigating Behavioral Plasticity in Insects: Application to Basic, Biomedical, and Agricultural Research

Description

The Proboscis Extension Response (PER) conditioning protocol, developed for the honey bee (Apis mellifera), provides an ecologically-relevant and easily quantifiable means for studying several different mechanisms of learning in many insect species.

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Date Created
  • 2014-09-08

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Testing Chronic and Acute Exposure of Selenium on Honey Bee Learning and Memory

Description

Selenium, a group 16 metalloid on the periodic table, is a necessary mineral for many organisms. Trace amounts of selenium are essential for normal development, antioxidant protein function, enzyme function,

Selenium, a group 16 metalloid on the periodic table, is a necessary mineral for many organisms. Trace amounts of selenium are essential for normal development, antioxidant protein function, enzyme function, and hormone regulation (Burden et al., 2016). However, when selenium is found in toxic amounts in organisms, it has been found to substitute for sulfur in proteins, which can be toxic to these animals, and cause oxidative stress (Quinn et al., 2007). Using the previous research done with acute exposure to organic and inorganic selenium compounds, we hypothesized that the inorganic sodium selenate would significantly decrease learning and memory recall for both chronic and acute exposure. We also hypothesized that the consumption of organic methylseleno-L-cysteine by honey bees would decrease learning and memory recall for both the chronic and acute exposure. We further hypothesized that protein carbonyl content would be increased due to oxidative damage caused by selenium in both the sodium selenate and the methylseleno-L-cysteine treatment groups, but that the inorganic selenium compound would increase the carbonyl content more than the methylseleno-L-cysteine. To run the experiments, three tents outside had two colonies in each tent. One tent contained the sodium selenate group, another had the sucrose control, and one contained the methylseleno-L-cysteine group. The treatment groups were fed selenium in their sucrose feeders. The first part of the experiment was training the bees by using proboscis extension response (PER) to teach them to extend their proboscis to the rewarded odor and not to the unrewarded odor. This was done by pairing the rewarded odor with a sucrose reward and not pairing it with the unrewarded odor. Then their short-term and long-term memory recall was tested. The second part of the experiment was checking for oxidative damage by measuring the protein carbonyl content in the bees. Three boxes were set up with the same three treatment groups as used in the tents. The treatment group bees were exposed to selenium in the sucrose feeders and in the pollen patties. After one week, the living bees were removed and frozen. They were then homogenized to extract protein. The first assay run was the protein content assay to establish a standard protein concentration for samples. Then a protein carbonyl assay was run, to determine the protein carbonyl content. Overall, the experiment found that exposure to selenium negatively impacted honey bees learning and memory recall significantly. Chronic exposure to the inorganic selenate reduced the bees' long-term memory abilities to differentiate between odors. With methylseleno-L-cysteine, it had no significant effect for the chronic exposure, but for the acute exposure, it had a significant impairment on their abilities to distinguish between the rewarded and unrewarded odors during conditioning. Our results showed that from our experiment there appeared to be no significant effect of selenium exposure on the increase of carbonylation content in the different treatment groups. This is most likely due to the fact the carbonyl content was not detectable because the protein concentration was low in the samples (approximately 3.5 mg/mL).

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Date Created
  • 2016-05

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The effect of Pristine fungicide on honey bee (Apis mellifera) taste and responsiveness to sucrose

Description

Honey bee (Apis mellifera) colonies have experienced substantial losses due to colony collapse disorder (CCD) since the first officially reported cases in 2006. Many factors have been implicated in CCD,

Honey bee (Apis mellifera) colonies have experienced substantial losses due to colony collapse disorder (CCD) since the first officially reported cases in 2006. Many factors have been implicated in CCD, including pests, pathogens, malnutrition, and pesticide use, but no correlation has been found between a single factor and the occurrence of CCD. Fungicides have received less research attention compared to insecticides, despite the fact that fungicide application coincides with bloom and the presence of bees. Pristine fungicide is widely used in agriculture and is commonly found as a residue in hives. Several studies have concluded that Pristine can be used without harming bees, but reports of brood loss following Pristine application continue to surface across the country. The primary objectives of this study were to determine whether Pristine causes an aversive gustatory response in bees and whether consumption of an acute dose affects responsiveness to sucrose. An awareness of how foragers interact with contaminated food is useful to understand the likelihood that Pristine is ingested and how that may affect bees' ability to evaluate floral resources. Our results indicated that Pristine has no significant effect on gustatory response or sucrose responsiveness. There was no significant difference between bee responses to Pristine contaminated sucrose and sucrose alone, and no significant effect of Pristine on sucrose responsiveness. These results indicate that honey bees do not have a gustatory aversion to Pristine. A lack of aversion means that honey bees will continue collecting contaminated resources and dispersing them throughout the colony where it can affect brood and clean food stores.

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Date Created
  • 2015-05