What's in endangered bat poop?

Raised of $2,800 Goal
Funded on 11/07/19
Successfully Funded
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  • Funded
    on 11/07/19



I will use DNA metabarcoding to identify prey insects and other arthropods in feces samples collected from wild bats. Bats were caught using harp traps and mist nets and sometimes poop while being held temporarily in cloth bags. After getting the DNA out of the poop using extraction kits, DNA “barcodes”, short regions of DNA that allow for identification of a species, will be amplified through PCR. After genetically indexing samples, pooling everything, and purifying the DNA, the precious results will be sent off for sequencing. After sequencing, I can begin cleaning the data and comparing each resulting barcode with barcoding databases to identify what prey were in the feces samples. The ecology of known prey items will give us clues as to what types of foraging habitat are important to each bat species. Guiding me in this process is collaborator, Dr. Eero Vesterinen, who has studied the diets of other bat species using this method.

「DNAメタバーコーディング法」を利用して、野生のコウモリから採集された糞サンプル中の被食昆虫や他の節足動物を特定します。コウモリはハープトラップとカスミ網を用いて捕獲します。コウモリが落ち着くために、一時的に巾着に入れておきますが、運が良ければ糞を出してくれます。抽出キットを使用して糞からDNAを取り出した後、種の識別を可能にするDNAの短い領域であるDNA「バーコード」をPCRによって増幅します。サンプルを遺伝的にインデックス付けし、すべてをプールし、DNAを精製した後、貴重な結果をシーケンスに送ります。シーケンス後、データの整理を開始し、得られた各バーコードをバーコードデータベースと比較して、糞サンプル中の獲物を識別します。獲物の生態は、各コウモリ種にとってはどんな生息地が採餌に重要であるかについての手がかりを与えてくれます。私の共同研究者である、Dr. Eero Vesterinenは、同じ方法を用いて他のコウモリ種の食性を解明したことがあり、私に助言を与えてくれています。


Figuring out what insects are in bat feces is not a simple task. Traditionally, this is done under a microscope and requires a high level of taxonomic expertise to identify the fragments of small body parts left after chewing and digestion. Additionally, soft-bodied insects, such as mosquitos, leave no visual evidence in the feces. I decided to use DNA metabarcoding to overcome both of these issues; even soft-bodied prey leave DNA, and my deficient knowledge of Okinawan arthropod pieces does not limit my ability to barcode their DNA. However, DNA metabarcoding is not free from challenges. Okinawa’s invertebrate fauna has not been thoroughly barcoded, so in some cases, I will only be able to identify a prey item to the genus, family, or order level. Additionally, I plan to consult local invertebrate specialists in order to assess the ecology of identified prey items and therefore infer the foraging habitats of my target bats.


Pre Analysis Plan

The first stage of analysis will follow standard DNA metabarcoding procedures. Once I have identified prey items as specifically as existing barcode databases allow, I will assess the types of prey that each bat species forages on. Prey types will be divided by taxonomic order, family, and genus, and also by their habitat (e.g. streams, understory, open fields). As I assess the results and consult with invertebrate specialists, I am likely to discover other prey attributes relevant to my research questions and will have to adapt to these. Based on wing shape, echolocation style, ecology of related species, and where I’ve previously found these bats, my hypotheses are that the Ryukyu tube-nosed bat specializes in gleaning insects off of surfaces in cluttered environments (including the ground!), while the Yanbaru whiskered bat hunts over forest streams catching prey on the wing. Assessing what types of habitat these bats prefer and how flexible they are in their foraging habits will help assess how vulnerable these bats are (more specialized species are more at risk) and what types of habitats to conserve. 



This project has not yet shared any protocols.