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Using Terrestrial Haematophagous Leeches to Enhance Tropical Biodiversity Monitoring Programmes in Bangladesh

Sarah R. Weiskopf
Department of Entomology and Wildlife Ecology, University of Delaware, Newark, DE, USA

Kyle P. McCarthy
Department of Entomology and Wildlife Ecology, University of Delaware, Newark, DE, USA

Michael Tessler
Richard Gilder Graduate School, American Museum of Natural History, New York, NY, USA

Hasan A. Rahman
Department of Entomology and Wildlife Ecology, University of Delaware, Newark, DE, USA

Jennifer L. McCarthy
Associate in Arts Program, University of Delaware, Wilmington, DE, USA

Rebecca Hersch
Sackler Institute for Comparative Genomics, American Museum of Natural History, New York, NY, USA

Mohammad Muzaffar Faisal
Creative Conservation Alliance, Dhaka, Bangladesh

Mark E. Siddall
Richard Gilder Graduate School, American Museum of Natural History, New York, NY, USA

Abstract/ Executive Summary:

Measuring mammal biodiversity in tropical rainforests is challenging, and methods that reduce effort while maximizing success are crucial for long-term monitoring programmes. Commonly used methods to assess mammal biodiversity may re -quire substantial sampling effort to be effective. Genetic methods are a new and important sampling tool on the horizon, but obtaining sufficient DNA samples can be a challenge.2. We evaluated the efficacy of using parasitic leeches Haemadipsa spp., as com -pared to camera trapping, to sample biodiversity. We collected 200 leeches from four forest patches in northeast Bangladesh, and identified recent vertebrate hosts using Sanger sequencing of the 16S rRNA gene extracted from each individual leech’s blood meals. We then compared these data to species data from camera trapping conducted in the same forest patches. 3. Overall, 41.9% of sequenced leeches contained amplifiable non-human mammal DNA. Four days of collecting leeches led to the identification of 12 species, com -pared to 26 species identified in 1,334 camera trap nights. 4. Synthesis and applications. After assessing the cost, effort and power of each technique, there are pros and cons to both camera trapping and leech blood meal analysis. Camera trapping and leech collection appear to be complementary approaches. When used together, they may provide a more complete monitoring tool for mammal biodiversity in tropical rainforests. Managers should consider adding leech collection to their biodiversity monitoring toolkit, as improved information will allow managers to create more effective conservation programmes.

Keywords: Blood meal sequencing, Camera trapping, DNA barcode, Haemadipsidae, Leech, Mammal biodiversity, Mitochondrial DNA, Non-invasive monitoring

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Start Date:

End Date:

Program Area: Conservation and Biodiversity

Commodity: Animal

Objectives:

Our objectives were to: (1) determine whether we can identify mammal species by sequencing leech blood meals, (2) determine whether leech size impacts amplification success rate of mammalian DNA, (3) compare performance of leech blood meal sequencing and camera trapping for detection of mammalian biodiversity, and (4) assess and compare costs and benefits of both methods.

Methodology:

Bordering the Indo- Burma biodiversity hotspot, Bangladesh is a small country with diverse flora and fauna, but also a rapidly diminishing tropical rainforest (Chowdhury & Koike, 2010). This study was con-ducted in northeast Bangladesh, a once highly forested area containing tropical evergreen and mixed evergreen forests. Most of the area has been deforested for roads, plantations and agriculture; the remaining forest is contained in 10 fragmented forest patches (Islam et al., 2013; Quazi & Ticktin, 2016). These patches are located between 24°4′ and 24°21′N and 91°15′ and 91°7′E, and range from 10 to 100 km². They are predominantly bordered by industrial plantations or rural settlements (Bangladesh Forest Department, 2012). The Forest Department manages most of the patches as “reserve forest,” indicating protected status with certain extraction activities permitted. The area also contains Satchari and Lawachara National Parks, and Rema- Kalenga Wildlife Sanctuary, where no extraction activities are allowed. eech collection Leeches were collected from the forest patches in October, 2015. We collected 50 leeches in each patch over four collection days, with one collection day per patch. At each patch, we picked five locations with previous camera trap data. We attempted to collect 10 leeches from each location, however if we were unable to find 10, we collected additional leeches at another nearby location to ensure we had 50 leeches per patch. We collected leeches by hand, using nitrile gloves to limit DNA contamination, and placed them into individual 1 mL test tubes filled with RNAlater® to preserve DNA for extended periods of time without refrigeration. For our analyses of BLAST results, we only included sequences with an e- value of less than e−30 to avoid low quality matches. We identified species using the top BLAST hit rather than a percent identity cutoff because we felt it was less subjective, as differences in inter- and intra- species similarities prevent determination of a universal cutoff (De Salle, Egan, & Siddall, 2005). We indicated whether the top hit had at least a 1% better percent identity match. This was assumed to be true if the first 100 matches were the same species. For the two blood meals where this was not the case, one was identified only to family, while the other was kept at species level since it is the only member of its genus in Bangladesh.Several blood meals matched species that do not occur in Bangladesh. The true species in these blood meals are likely missing sequences in GenBank. We therefore reported these only to family but counted them as separate species in the analysis, as they were separate taxonomic units than those encountered in other leeches and may have been identifiable if we sequenced additional genes or bolstered the database by sequencing museum specimens. Our species identifications should be considered with caution, as we sequenced only one gene. However, because the exact identity of the species would not change our conclusions, we did not sequence additional genes.

Source of Information: J Appl Ecol. 2018; 55: 2071–2081

Article Link (Online Journal):

Funding Source:

1.

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Total Budget (Tk.) :

Achievement/Findings:

Monitoring programmes are desperately needed in the tropics, where biodiversity and threats to biodiversity are high and data are limited (Burton, 2012). In order to be effective and sustainable in the long term, monitoring programmes must be supported financially, politically and logistically (Lindenmayer, 1999). Using leech blood meals to monitor biodiversity is potentially cheaper and more efficient than camera trapping for large sample sizes. Even with as few as 200 leeches, we extended the species list from an extensive camera trapping effort. However, further studies are needed to determine whether larger leech sample sizes can detect comparable levels of species richness as camera trapping and what biases may be associated with the method. In this study, we presented some of the cost- performance trade- offs of camera trapping and leech blood meal sequencing, allowing managers to make more informed decisions about which technique to utilize. Camera trapping and other survey methods require independent corroboration, which can be provided rapidly and non- invasively by leech blood meal analysis. Managers should consider using the two methods together to improve the efficiency and capacity of monitoring programmes. Sustainable monitoring programmes and improved under standing of species diversity will allow researchers to create more effective conservation plans.

Knowledge Management: Journal