Fajardo, Christopher

Discovery and Characterization of Honey Bee Larvae Gut Microbiota

Faculty Mentor: John Chaston, Plant & Wildlife Sciences

Introduction

Honey bees are currently plagued by an infection known as American Foulbrood (AFB). This disease is caused by the bacterium Paenibacillus larvae. It is carried by adult Honey bees and becomes an active infection once inside the digestive tract of a Honey bee larvae. Current treatment of this disease is a broad-spectrum antibiotic with negative effects on both the Honey bee and the environment. Research is currently being conducted with the hope to introduce a bacteriophage therapy to the market. This approach will be specific to Paenibacillus larvae without the worries of the current treatments. Our focus is to provide an accurate microbiota1 of Honey bee larvae through its development. This will allow researchers to test their therapy’s toxicity on beneficial bacteria.

Materials and Methods

Local Beekeepers graciously provided us with Honey bee larvae in various stages of development for our study. They were transported to our lab in a cooler that maintained an environment of 35° C and 40% – 50% humidity to preserve the larvae until dissection.

Dissection was performed in sterile 1x PBS solution with the aid of a dissection scope (See Fig. 1). After the digestive tract of the larvae was isolated and removed. It was put on ice until transfer to a -40° C freezer for storage until DNA isolation could be performed.

DNA isolation was performed on the larval digestive tract using standard methods to preserve the microbiota. This DNA will be sequenced on the BYU HiSeq 2500 Illumina sequencer once a run becomes available.

Once the sequencing is finished. We will use software to analyze the data to provide us with an accurate view of a Honey bee larvae ‘s microbiota. Giving us specific species of bacteria and in what quantities.

Results

Specific data on the Honey Bee microbiota is unavailable at the time of this paper’s publication due to our late submission of our DNA samples. A note of interest, however, is the variety of the larvae samples from the various locations we collected samples. We attempted to take as broad of a sample as we could. Even with our efforts, variations in larvae color and size were apparent across sample locations. As well as the sources of food the Honey bees could collect form. Our hopes are that this will reflect in our subsequent analysis.

Conclusion

A microbiota tells a large story of an organism’s health and well-being. Considering the large impact that honeybees have on an ecosystem, not to mention our economy. A well-defined microbiota will help in current research and in future attempts at preserving this vital species.

Citations

1 For a great paper on the benefits of microbiota research. I recommend: Guinane CM, Cotter PD. Role of the gut microbiota in health and chronic gastrointestinal disease: understanding a hidden metabolic organ. Therapeutic Advances in Gastroenterology. 2013;6(4):295-308. doi:10.1177/1756283X13482996.