Endangered Nectar-Feeding Bat Detected by Environmental DNA on Flowers
<i>Leptonycteris nivalis</i> (the Mexican long-nosed bat) is an endangered nectar-feeding bat species that follows “nectar corridors” as it migrates from Mexico to the southwestern United States. Locating these nectar corridors is key to their conservation and may be possible using envir...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-11-01
|
| Series: | Animals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2615/12/22/3075 |
| _version_ | 1827645240900583424 |
|---|---|
| author | Faith M. Walker Daniel E. Sanchez Emma M. Froehlich Emma L. Federman Jacque A. Lyman Meagan Owens Kristen Lear |
| author_facet | Faith M. Walker Daniel E. Sanchez Emma M. Froehlich Emma L. Federman Jacque A. Lyman Meagan Owens Kristen Lear |
| author_sort | Faith M. Walker |
| collection | DOAJ |
| description | <i>Leptonycteris nivalis</i> (the Mexican long-nosed bat) is an endangered nectar-feeding bat species that follows “nectar corridors” as it migrates from Mexico to the southwestern United States. Locating these nectar corridors is key to their conservation and may be possible using environmental DNA (eDNA) from these bats. Hence, we developed and tested DNA metabarcoding and qPCR eDNA assays to determine whether <i>L. nivalis</i> could be detected by sampling the agave flowers on which it feeds. We sampled plants with known bat visitations in the Sierra Madre Oriental in Laguna de Sanchez (LS), Nuevo León, Mexico, and in the Chisos Mountains in Big Bend National Park, TX, USA (CB). A total of 13 samples included both swabs of agave umbels and cuttings of individual flowers. DNA metabarcoding was performed as a PCR multiplex that targeted bats (SFF-COI), arthropods (ANML-COI), and plants (ITS2 and rbcL). We targeted arthropods and plants in parallel with bats because future metabarcoding studies may wish to examine all the pollinators and plants within the nectar corridor. We developed and tested the sensitivity and specificity of two qPCR assays. We found that both DNA metabarcoding and qPCR were highly successful at detecting <i>L. nivalis</i> (11 of 13 for DNA metabarcoding and 12 of 13 for qPCR). Swabs and flower cuttings and both qPCR assays detected the species over four replicates. We suggest that <i>L. nivalis</i> leaves substantial DNA behind as it forages for nectar. We also suggest that future studies examine the time since sampling to determine its effect on detection success. The DNA metabarcoding multiplex will be useful for parallel questions regarding pollination ecology, while, with further testing, the qPCR assays will be effective for large-scale sampling for the detection of migration corridors and foraging areas. This work may be relevant to other nectar-feeding bat species, which can likely be detected with similar methodologies. |
| first_indexed | 2024-03-09T18:32:39Z |
| format | Article |
| id | doaj.art-d3e580174a0146dc9a9155b6f09a76c8 |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-09T18:32:39Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-d3e580174a0146dc9a9155b6f09a76c82023-11-24T07:27:53ZengMDPI AGAnimals2076-26152022-11-011222307510.3390/ani12223075Endangered Nectar-Feeding Bat Detected by Environmental DNA on FlowersFaith M. Walker0Daniel E. Sanchez1Emma M. Froehlich2Emma L. Federman3Jacque A. Lyman4Meagan Owens5Kristen Lear6Bat Ecology & Genetics Lab, School of Forestry, Northern Arizona University, Flagstaff, AZ 86011, USABat Ecology & Genetics Lab, School of Forestry, Northern Arizona University, Flagstaff, AZ 86011, USABat Ecology & Genetics Lab, School of Forestry, Northern Arizona University, Flagstaff, AZ 86011, USABat Ecology & Genetics Lab, School of Forestry, Northern Arizona University, Flagstaff, AZ 86011, USABat Ecology & Genetics Lab, School of Forestry, Northern Arizona University, Flagstaff, AZ 86011, USABat Ecology & Genetics Lab, School of Forestry, Northern Arizona University, Flagstaff, AZ 86011, USAIntegrative Conservation and Forestry & Natural Resources, University of Georgia, Athens, GA 30602, USA<i>Leptonycteris nivalis</i> (the Mexican long-nosed bat) is an endangered nectar-feeding bat species that follows “nectar corridors” as it migrates from Mexico to the southwestern United States. Locating these nectar corridors is key to their conservation and may be possible using environmental DNA (eDNA) from these bats. Hence, we developed and tested DNA metabarcoding and qPCR eDNA assays to determine whether <i>L. nivalis</i> could be detected by sampling the agave flowers on which it feeds. We sampled plants with known bat visitations in the Sierra Madre Oriental in Laguna de Sanchez (LS), Nuevo León, Mexico, and in the Chisos Mountains in Big Bend National Park, TX, USA (CB). A total of 13 samples included both swabs of agave umbels and cuttings of individual flowers. DNA metabarcoding was performed as a PCR multiplex that targeted bats (SFF-COI), arthropods (ANML-COI), and plants (ITS2 and rbcL). We targeted arthropods and plants in parallel with bats because future metabarcoding studies may wish to examine all the pollinators and plants within the nectar corridor. We developed and tested the sensitivity and specificity of two qPCR assays. We found that both DNA metabarcoding and qPCR were highly successful at detecting <i>L. nivalis</i> (11 of 13 for DNA metabarcoding and 12 of 13 for qPCR). Swabs and flower cuttings and both qPCR assays detected the species over four replicates. We suggest that <i>L. nivalis</i> leaves substantial DNA behind as it forages for nectar. We also suggest that future studies examine the time since sampling to determine its effect on detection success. The DNA metabarcoding multiplex will be useful for parallel questions regarding pollination ecology, while, with further testing, the qPCR assays will be effective for large-scale sampling for the detection of migration corridors and foraging areas. This work may be relevant to other nectar-feeding bat species, which can likely be detected with similar methodologies.https://www.mdpi.com/2076-2615/12/22/3075environmental DNAeDNAChiropterapollinationhigh-throughput nucleotide sequencingDNA metabarcoding |
| spellingShingle | Faith M. Walker Daniel E. Sanchez Emma M. Froehlich Emma L. Federman Jacque A. Lyman Meagan Owens Kristen Lear Endangered Nectar-Feeding Bat Detected by Environmental DNA on Flowers Animals environmental DNA eDNA Chiroptera pollination high-throughput nucleotide sequencing DNA metabarcoding |
| title | Endangered Nectar-Feeding Bat Detected by Environmental DNA on Flowers |
| title_full | Endangered Nectar-Feeding Bat Detected by Environmental DNA on Flowers |
| title_fullStr | Endangered Nectar-Feeding Bat Detected by Environmental DNA on Flowers |
| title_full_unstemmed | Endangered Nectar-Feeding Bat Detected by Environmental DNA on Flowers |
| title_short | Endangered Nectar-Feeding Bat Detected by Environmental DNA on Flowers |
| title_sort | endangered nectar feeding bat detected by environmental dna on flowers |
| topic | environmental DNA eDNA Chiroptera pollination high-throughput nucleotide sequencing DNA metabarcoding |
| url | https://www.mdpi.com/2076-2615/12/22/3075 |
| work_keys_str_mv | AT faithmwalker endangerednectarfeedingbatdetectedbyenvironmentaldnaonflowers AT danielesanchez endangerednectarfeedingbatdetectedbyenvironmentaldnaonflowers AT emmamfroehlich endangerednectarfeedingbatdetectedbyenvironmentaldnaonflowers AT emmalfederman endangerednectarfeedingbatdetectedbyenvironmentaldnaonflowers AT jacquealyman endangerednectarfeedingbatdetectedbyenvironmentaldnaonflowers AT meaganowens endangerednectarfeedingbatdetectedbyenvironmentaldnaonflowers AT kristenlear endangerednectarfeedingbatdetectedbyenvironmentaldnaonflowers |