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Title:   Author  item
                                            PHILLIPP, ENGEL - Yale Universityitem
                                            James, Rosalinditem
                                            KOGA, RYUICHI - National Institute Of Advanced Industrial Science And Technology (NIAIST)item
                                            KWONG, WALDON - Yale Universityitem
                                            MCFREDERICK, QUINN - University Of Texasitem
                                            MORAN, NANCY - Yale University
  Submitted to: Journal of Apicultural Research
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 5/22/2013
                                    Publication  Date: 9/1/2013
                                    Citation: Phillipp, E., James, R.R., Koga, R., Kwong, W., Mcfrederick, Q., Moran, N. 2013. Standard methods for research on Apis mellifera gut symbionts. Journal of Apicultural Research. 52(4):1-23.Interpretive Summary: This paper describes methods for studying symbiotic gut microbes in honey bees, and is part of a series describing standard methods for honey bee research. Gut microbes can play an important role in digestion, disease resistance, and the general health of animals, in general, but little is known about the biology of gut symbionts in honey bees. Traditional microbiological culturing techniques have been used in the past to describe the honey bee gut microflora.  However, such methods underestimate the true diversity of microorganisms in most environments because many microbes cannot be artificially cultured, or require specialized conditions that may not have been provided by the investigator. We describe methods which utilize DNA sequencing technologies to survey microorganisms present in honey bee guts; methods for using fluorescent in situ hybridization (a method which causes selected microorganisms to fluoresce under the microscope) to identify where in the gut certain species occur; and methods for culturing some honey bee gut symbionts so that the biology of these microorganisms can be studied in more detail.  We hope that the methods described will help other researchers advance the state of knowledge regarding honey bee gut symbionts, an intriguing area of research.
                                Technical Abstract: Gut microbes can play an important role in digestion, disease resistance, and the general health of animals, but little is known about the biology of gut symbionts in Apis mellifera. This paper is part of a series on honey bee research methods, providing protocols for studying gut symbionts. We describe here non-culture-based approaches based on NGS sequencing, as these methods have greatly improved our capabilities when it comes to identifying the microbial communities associated with honey bees.  We also describe florescent in situ hybridization (FISH) microscopy, which allows a visual examination of the microenvironments where particular microbes occur. Culturing methods are also described, as they allow the researcher to isolate particular bacteria of interest for further study or gene identification, and enable the assignment of particular functions to particular gut community members.  Metagenomic methods were not included because this is a rapidly evolving approach to environmental microbiology, and standard methodologies are still under development at this time.  We hope the methods we have included will help others advance the state of knowledge regarding bee gut symbionts and the role they play in honey bee health.


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