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UNLV Theses, Dissertations, Professional Papers, and Capstones
12-15-2019
The Effect of Pleistocene Glacial - Interglacial Cycles on Patterns The Effect of Pleistocene Glacial - Interglacial Cycles on Patterns
of Genetic Diversity and Differentiation of Populations of of Genetic Diversity and Differentiation of Populations of
Leptodactylus Albilabris (White-Lipped Frog) in the Puerto Rican Leptodactylus Albilabris (White-Lipped Frog) in the Puerto Rican
Bank Bank
Andre Nguyen
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Part of the Biology Commons
Repository Citation Repository Citation Nguyen, Andre, "The Effect of Pleistocene Glacial - Interglacial Cycles on Patterns of Genetic Diversity and Differentiation of Populations of Leptodactylus Albilabris (White-Lipped Frog) in the Puerto Rican Bank" (2019). UNLV Theses, Dissertations, Professional Papers, and Capstones. 3830. http://dx.doi.org/10.34917/18608737
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THE EFFECT OF PLEISTOCENE GLACIAL - INTERGLACIAL CYCLES ON PATTERNS
OF GENETIC DIVERSITY AND DIFFERENTIATION OF POPULATIONS OF
LEPTODACTYLUS ALBILABRIS (WHITE-LIPPED FROG)
IN THE PUERTO RICAN BANK
By
Andre Nguyen
Bachelor of Science – Biological Sciences San Diego State University
2015
A thesis submitted in partial fulfillment of the requirements for the
Master of Science – Biological Sciences
School of Life Sciences College of Sciences
The Graduate College
University of Nevada, Las Vegas December 2019
ii
Thesis Approval
The Graduate College The University of Nevada, Las Vegas
October 02, 2019
This thesis prepared by
Andre Nguyen
entitled
The Effect of Pleistocene Glacial - Interglacial Cycles on Patterns of Genetic Diversity and Differentiation of Populations of Leptodactylus Albilabris (White-Lipped Frog) in the Puerto Rican Bank
is approved in partial fulfillment of the requirements for the degree of
Master of Science – Biological Sciences School of Life Sciences
Javier Rodriguez, Ph.D. Kathryn Hausbeck Korgan, Ph.D. Examination Committee Chair Graduate College Dean Dennis Bazylinski, Ph.D. Examination Committee Member Mira Han, Ph.D. Examination Committee Member Tereza Jezkova, Ph.D. Examination Committee Member Matthew Lachniet, Ph.D. Graduate College Faculty Representative
iii
Abstract
The historical distributional shifts of various species have been attributed to the glacial-
interglacial cycles of the Pleistocene Epoch (2.6 mya – 10,000 ya). Sea level changes caused by
Pleistocene glacial-interglacial cycles impacted the total area of landmasses, particularly in
island systems. Cooler, glacial periods increased emergent (subaerial) island area via lower sea
levels. Adjacent islands may have become connected through emergent land bridges, resulting in
a larger subaerial landmass. Warmer interglacial periods led to higher sea levels, which in turn
reduced island area, and in some cases, submerged land bridges, fragmenting larger islands into
separate, smaller units. The Puerto Rican Bank (PRB) is an island system in the eastern
Caribbean Sea consisting of more than 180 islands, islets, and cays. The largest islands of the
PRB are Puerto Rico, Vieques, Culebra, Saint Thomas, Saint John, Jost Van Dyke, Tortola,
Virgin Gorda, and Anegada (Saint Thomas, Saint John, and Saint Croix form part of the United
States Virgin Islands, but Saint Croix lays in its own bank, and has not had a land connection to
the PRB.) Changing sea levels influenced the degree of land exposure and connectivity across
the PRB. Lower sea levels revealed the shelf, and connected all the present-day islands of the
PRB into a single landmass. Conversely, higher sea levels submerged the shelf, fragmenting the
PRB into numerous islands, as in its current configuration. The repeated episodes of island
connectivity and isolation of the PRB may have left a genetic imprint on native species. I relied
on mitochondrial DNA and nuclear genomic sequences to assess whether historic sea levels
associated with Pleistocene glacial-interglacial cycles have shaped patterns of present-day
genetic diversity in Leptodactylus albilabris (White-lipped Frog) in the PRB. I tested four
specific hypotheses. (i) The boundaries of the five physiographic regions of Puerto Rico present
iv
a physical or ecological barrier to migration among L. albilabris from the different regions.
(ii) There is a positive correlation between geographic and genetic distance, or “isolation by
distance” (IBD), among populations of L. albilabris across the PRB. (iii) Puerto Rico is the
ancestral area of L. albilabris. (iv) Because Saint Croix has not had a land connection to the
PRB, the origin of the Saint Croix populations of L. albilabris is either the result of natural
colonization or human-mediated introduction, or of in situ evolution of the species on Saint
Croix. My findings are partially supported, nuclear genomic analyses revealed distinct genetic
groups corresponding to the Cordillera Central and the rest of Puerto Rico. I did not detect an
association between geographic and genetic distance among populations of L. albilabris across
the PRB, which suggests that this frog does not exhibit IBD across its distribution. Genetic
diversity and heterozygosity estimates from mitochondrial and nuclear genomic DNA data sets
suggested that Puerto Rico may be the ancestral area of L. albilabris. Both mitochondrial and
nuclear genomic analyses suggested that the Saint Croix populations of L. albilabris are the
result of introduction. The Saint Croix populations of L. albilabris are the least genetically
diverse of all populations of the frog, and are genetically very similar to the Saint Thomas
populations, suggesting that Saint Thomas is the source of the Saint Croix L. albilabris. My
study characterized the genetic architecture of L. albilabris across its distribution in the PRB, and
contributed to our understanding of how historical changes in Pleistocene sea levels have shaped
patterns of present-day genetic diversity in island systems.
v
Acknowledgements
I thank my graduate committee members: Dr. Javier Rodríguez (School of Life Sciences),
for accepting me as a student and helping me cultivate my interest and knowledge in
biodiversity; Dr. Dennis Bazylinski (School of Life Sciences), for providing resources that taught
me new skills that I had not had opportunities to develop before coming to UNLV; Dr. Mira Han
(School of Life Sciences), for guiding me in the right direction with bioinformatics data analysis
and introducing me to a different aspect of the biological sciences, Dr. Matthew Lachniet
(Department of Geoscience), for providing expertise about aspects of historic global climate
relevant to this project; and Dr. Tereza Jezkova (Department of Biology, Miami University,
Ohio), for guidance with the computationally intensive analyses used in this project. I also thank
Dr. Viviana Morillo López (formerly at UNLV’s School of Life Sciences and presently at
Marine Biological Laboratory, University of Chicago), for taking the time to mentor me and
teach me laboratory techniques; and Dr. Richard Tillett (Ecology, Evolution and Conservation
Biology Graduate Program, University of Nevada, Reno), for providing me with hands-on
bioinformatics training that I have developed a great interest for. I also acknowledge the
committee members from the School of Life Sciences Aquatic Biology Endowment for granting
me funding to conduct fieldwork, and the Nevada INBRE service awards for funding my
bioinformatics training. These funding opportunities have provided me opportunities to become a
more experienced and well-rounded biologist. Many thanks to my fellow graduate students, and
friends in the School for being a part of my experience at UNLV. Special thanks to my
girlfriend, Shannon Aurigemma, for her never-ending support. Finally, I thank my family
Phuong, Dung, and Andrea Nguyen, for their love and support is the foundation that I stand on.
vi
Table of Contents
Abstract .......................................................................................................................................... iii Acknowledgements ......................................................................................................................... v Table of Contents ........................................................................................................................... vi List of Tables ............................................................................................................................... viii List of Figures ................................................................................................................................ ix CHAPTER 1: Introduction ............................................................................................................. 1 CHAPTER 2: Materials and Methods ............................................................................................ 6
Taxon sampling ........................................................................................................................... 6 Mitochondrial DNA - Laboratory Methods ................................................................................ 6 Mitochondrial DNA - Analyses .................................................................................................. 7 Double-digest restriction site associated DNA-sequencing - Laboratory Methods ................... 8 Double-digest restriction site associated DNA-sequencing - Analyses .................................... 10
CHAPTER 3: Results ................................................................................................................... 12 Bayesian phylogenetic analysis of mitochondrial DNA ........................................................... 12 Haplotype network of mitochondrial DNA .............................................................................. 13 Haplotype diversity and nucleotide diversity of mitochondrial DNA ...................................... 14 Population structuring and association between geographic and genetic distance calculated
using mitochondrial DNA ......................................................................................................... 15 Double-digest restriction site associated DNA-sequencing SNP-filtering ............................... 16 Population genomic analyses – heterozygosity and phylogenomic structure ........................... 17
vii
CHAPTER 4: Discussion .............................................................................................................. 19 The effect of Puerto Rico’s physiography and Pleistocene climate on patterns of genetic
variation in Leptodactylus albilabris ........................................................................................ 19 Isolation by distance of Leptodactylus albilabris populations across the Puerto Rican Bank . 22 Ancestral area of Leptodactylus albilabris ............................................................................... 23 Origins of the Saint Croix populations of Leptodactylus albilabris ......................................... 25 Systematic status of Leptodactylus dominicensis ..................................................................... 27 Concluding remarks .................................................................................................................. 28
References ..................................................................................................................................... 92 Curriculum Vitae ........................................................................................................................ 109
viii
List of Tables
Table 1. Sample list....................................................................................................................... 29 Table 2. Unique Mitochondrial DNA haplotypes by phylogroup ................................................ 54 Table 3. Mitochondrial DNA haplotypes by population .............................................................. 56 Table 4. Mitochondrial DNA genetic diversity statistics............................................................. 59 Table 5. Pairwise fixation index values among populations of Leptodactylus albilabris ............ 63 Table 6. Single nucleotide polymorphism heterozygosity statistics ............................................. 78
ix
List of Figures
Figure 1. Map of the 42 sampling localities for Leptodactylus albilabris .................................... 80 Figure 2. Bayesian inferrence phylogenetic tree of unique cytochrome b haplotypes ................. 82 Figure 3. Haplotype network map of cytochrome b haplotypes ................................................... 85 Figure 4. Individual sample ancestry matrix and relative population ancestry proportions
generated from single nucleotide polymorphisms ........................................................................ 87 Figure 5. Configurations of the Puerto Rican Bank at 10 meter intervals (0 m – -60 m) ............. 89
1
CHAPTER 1: Introduction
Resolving the geographic distribution of genetic lineages, that is, the phylogeographic
patterns of a species, can provide insight into the historical events that have shaped modern
species distributions. Making inferences that incorporate natural, historical, and recent processes
requires integrating data from various scientific fields (Kidd & Ritchie, 2006; Hassine et al.,
2016) such as geology, population genetics, and ecology. The climate and sea level oscillations
that occurred throughout the Pleistocene Epoch (2.6 mya – 10,000 ya) are a well-documented
example of intermittent episodes that influenced the demographic history of biota (Hewitt, 1996;
Leal et al., 2016; Gehara et al., 2017; Silva et al., 2017; Parvizi et al., 2018; Potter et al., 2018;
Hyseni & Garrick, 2019). Climate may have impacted the vagility of organisms, whereas sea
level changes affected land area configurations globally. Land area configuration changes via
eustatic sea levels (i.e. uniformly global variation of sea level) facilitated population isolations
through habitat fragmentations and land area contractions. Through increased land exposure,
eustatic sea levels paradoxically also led to range expansion and secondary contact of previously
separated populations (Hewitt, 1996, 2000, 2004; Petit et al., 2003; Krysko et al., 2016).
The sea level changes of the Pleistocene Epoch resulted from alternating episodes of
global cooling and warming, or glacial-interglacial cycles. The extensive, cooler glacial periods
froze ocean water as massive ice sheets, lowering sea levels for up to ca. -134 m (Rohling et al.,
1998; Bintanja et al., 2005; Lambeck et al., 2014), compared to present-day sea levels. On the
contrary, the shorter, warmer interglacial periods of the Pleistocene Epoch led to smaller ice
volumes and higher sea levels (Weigelt et al., 2016). The demographic consequences of these sea
level changes can be inferred using phylogenetic and population genetic methods that detect
2
molecular signals in present-day taxa (e.g. Davison & Chiba, 2008; Barrow et al., 2017;
Moriyama et al., 2018; Deli et al., 2019).
Islands are considered natural platforms of evolution (Losos & Ricklefs, 2009).
Populations on islands are enclosed by abiotic boundaries, presenting opportunities for
evolutionary processes to occur undisturbed from non-native biotic forces (Slatkin, 1987; Wade
& McCauley, 1988; Whitlock & McCauley, 1990; Reynolds et al., 2017). Processes such as
immigration, extinction, population differentiation, and speciation can occur within individual
islands and archipelagos (Clouse et al., 2015; Fernández-Palacios et al., 2016; Hirano et al.,
2019), contributing to the complexities of evolutionary history of island species. During
interglacial periods of the Pleistocene, rising sea levels submerged land bridges that connected
islands systems. Rising sea levels thus caused islands to fragment into smaller, more isolated
units. Island fragmentation may have led to population isolation, which can influence the genetic
architecture of a species. On the other hand, the lower sea levels of glacial periods increased
island area by exposing submerged land bridges (Weigelt et al., 2016). Newly adjoined islands
presented opportunities for colonization or reconnection of previously isolated populations (Garg
et al., 2018). Sea level changes of the Pleistocene also impacted marine currents, and therefore
influenced the paths of organisms on flotsam (Fernández-Palacios et al., 2016), altering paths of
colonization and genetic exchange between island populations.
The Puerto Rican Bank (PRB) is a 350 km long island system located in the eastern
Caribbean Sea. This group of islands consists of Puerto Rico and the “Eastern Islands:” Vieques,
Culebra, the United States Virgin Islands (Saint Thomas and Saint John), the British Virgin
Islands (Jost Van Dyke, Tortola, Virgin Gorda, Anegada), and ca. 180 associated islands, islets
and cays (Barker et al., 2012). This archipelago is connected by a submerged shelf less than 120
3
m below current sea level (Heatwole & MacKenzie, 1967; Hedges, 1999; Renken et al.,
2002). Glacial periods of the Pleistocene revealed the shelf, and united all the modern islands of
the PRB into a contiguous landmass approximately twice the area (ca. 18,000 km2) of Puerto
Rico, the largest island of the archipelago (Thomas, 1999; Figure 1). Politically, Saint Croix is
part of the United States Virgin Islands, but this island lays in its own bank and has not had a
land connection to the PRB (Heatwole & MacKenzie, 1967).
Leptodactylus albilabris (White-lipped frog; Gunther, 1859) is a native, widespread
species in the PRB. The maximum reported snout-to-vent length of adults is 49 mm (Schwartz &
Henderson, 1991; Hedges & Heinicke, 2007). This frog typically occurs in habitats with standing
fresh water, where adult males can be found in dense aggregations advertising for mates (López
et al., 1988). Adult females deposit eggs in foam nests; rain may wash away the eggs from the
nests into nearby ponds in which free-living aquatic larva (i.e. tadpoles) hatch (Dent, 1956;
Schwartz & Henderson, 1991; Joglar, 2005; Hedges & Heinicke, 2007). Amphibians are
excellent models to investigate phylogeographic patterns in island systems. Their relatively low
rates of dispersal and great climatic and environmental sensitivity present opportunities to
explore the impact of glacial cycles on biogeography and genealogies (Zeisset & Beebee, 2008;
Garcia-Porta et al., 2012). I characterized the population genetic architecture of L. albilabris
throughout the PRB to investigate the potential effect of eustatic sea level changes during the
Pleistocene on the population genetic structure of this frog.
I predicted that the boundaries between the physiographic regions of Puerto Rico restrict
gene flow among populations of L. albilabris from different areas of the island. Puerto Rico has
five major physiographic regions: (i) the Cordillera Central, (ii) Sierra de Cayey, (iii) Sierra de
Luquillo, (iv) Cuchilla de Pandura, and (v) the Lowlands (Hedges, 1999). The boundaries of
4
these areas may present a physical and/or ecological barrier to dispersal of L. albilabris across
physiographic regions. If this hypothesis is correct, population genetic analyses would reveal
distinctive genetic groups (i.e. clades) corresponding to these physiographic regions.
The population genetic architecture of L. albilabris across the PRB may reflect “isolation
by distance” (IBD). This pattern is exhibited when geographic distance facilitates gradual genetic
differentiation across a species’ distribution, so that there is a positive relationship between
geographic distance and genetic divergence between populations (Wright, 1943; Kimura, &
Weiss, 1964; de Aguiar et al., 2009; Baptestini et al., 2013). If this scenario is accurate, Puerto
Rican populations of L. albilabris should display a greater degree of genetic divergence from the
more distant populations of the British Virgin Islands, compared to populations from the closer
United States Virgin Islands. I also expect the intermediately located United States Virgin
Islands populations of L. albilabris to exhibit genetic similarities with both Puerto Rican and
British Virgin Islands populations of this frog.
I hypothesized that Puerto Rico, the largest island of the PRB, is the ancestral area of L.
albilabris. If this hypothesis is correct, population genetic analyses should show genetic markers
from Puerto Rican populations located at the base of the L. albilabris species phylogeny, and
Puerto Rican populations having greater genetic diversity than the Eastern Islands populations.
Alternatively, analyses may detect greater genetic diversity and basal positioning of genetic
markers from the Eastern Islands populations of L. albilabris, implying that the frog originated
on the Eastern Islands and dispersed westward.
As stated earlier, Saint Croix (U.S. Virgin Islands) lays in its own bank and has not had a
land connection to the PRB (Heatwole & MacKenzie, 1967). Because L. albilabris occurs on
Saint Croix, I hypothesized that the frog colonized Saint Croix from the PRB, naturally or via
5
human-mediated introduction(s), either accidentally (Perry et al., 2006) or deliberately (cf.
Barker & Rodríguez-Robles, 2017). If this prediction is correct, genetic markers from Saint
Croix should be identical to, or derived from L. albilabris populations in the PRB. On the
contrary, greater levels of genetic variation in Saint Croix L. albilabris may indicate that the frog
evolved in situ on the island, from which it colonized the PRB. In this scenario, oceanic currents
could have carried frogs on flotsam to the PRB. If Saint Croix is the ancestral area of L.
albilabris, genetic markers from Saint Croix individuals would be positioned basally on the L.
albilabris species phylogeny.
In 1923, Cochran described Leptodactylus dominicensis based on a single specimen from
Hispaniola (Cochran, 1923), an island (ca. 76,200 km2) located west of Puerto Rico. In her
original description and in a later systematic monograph on the herpetology of Hispaniola
(Cochran, 1941), Cochran stated that L. dominicensis, known only from a small area in
northeastern Hispaniola, is closely related to L. albilabris. Heyer (1978) synonymized L.
dominicensis with L. albilabris, a taxonomic rearrangement supported by Hedges and Heinicke
(2007) and by de Sá et al. (2014), based on L. dominicensis’ marked morphological and genetic
similarity to L. albilabris. I relied on a denser sampling of L. albilabris across the PRB to assess
the systematic position of L. dominicensis.
6
CHAPTER 2: Materials and Methods
Taxon sampling
I collected 322 individuals of L. albilabris from 42 localities across eight islands: Puerto
Rico (n = 243), Vieques (n = 11), Culebra (n = 10), Saint Thomas (n = 12), Saint John (n = 10),
Saint Croix (n = 18), Jost Van Dyke (n = 11), and Tortola (n = 7) (Figure 1; Table 1). Sample
size varied from 5 – 17 frogs per locality.
Mitochondrial DNA - Laboratory Methods
I extracted genomic DNA from tissue samples preserved in 95% ethanol (liver, heart,
stomach, muscle) with the DNeasy Blood and Tissue Kit (Qiagen Inc., Valencia, CA). I used the
primers CBL1 (TCTGCYTGATGAAAYTTTGG) and CBH15
(ACTGGTTGDCCYCCRATYAK; Hedges & Heinecke, 2007) to amplify 900 base pairs (bp) of
the mitochondrial DNA marker cytochrome b (cyt b). I carried out PCR reactions in 50 μl
volumes consisting of 2 μl of template DNA, 0.5 μl of each primer (10 μM), 25 μl of GoTaq®
Green Master Mix (Promega Corp., Madison, WI), and 22 μl of ddH2O. I denatured DNA at
95°C for 2.5 min, then performed 32 cycles of amplification as follows: denaturation at 95°C for
1 min, annealing at 51°C for 1 min, and extension at 72°C for 1.5 min, followed by a final
extension at 72°C for 10 min. I cleaned the PCR products with the Zymo Genomic DNA Clean
and ConcentratorTM kit (Zymo Research Corporation, Irvine, CA, USA). Sequencing of cyt b
was performed at the Genomics Core Facility of the University of Nevada, Las Vegas, with the
same primers used for amplification.
7
I aligned the sequences using SEQUENCHER 5.3 (Gene Codes Corporation, Ann Arbor,
MI), and verified the alignment visually. Missing data were treated as a fifth character state at the
particular site. The final mitochondrial DNA (mtDNA) data set consisted of 828 bp of cyt b.
I used DnaSP 6.10.04 (Rozas et al., 2017) to collapse the 322 cyt b sequences to 140 unique
haplotypes. I incorporated cyt b sequence data from one specimen of L. dominicensis
(GenBank accession number EF 091393.1) into the ingroup.
Mitochondrial DNA - Analyses
Prior to conducting phylogenetic analyses, I determined best fitting models of nucleotide
substitution for each codon position with jModelTest 2.1.10 (Darriba et al., 2012; Guindon &
Gascuel, 2003). The Akaike Information Criteria identified K80 (for the first and third codon
positions) (Kimura, 1980), and JC (for the second codon position) (Jukes & Cantor, 1969) as the
most appropriate models. I inferred phylogenetic relationships among the unique cyt b
haplotypes using Bayesian Inference (BI) criteria, as implemented in the program MrBayes 3.2.6
(Ronquist et al., 2012). I produced posterior probability distributions by allowing four Monte
Carlo Markov Chains to proceed for 40 million generations each, with samples taken every 100
generations, a procedure that yielded 120,002 trees. After visual evaluation, I discarded the first
25% of trees as “burn-in” samples, and combined the remaining samples to estimate tree
topology, posterior probability values, and branch lengths. I confirmed convergence of all
parameters in Tracer 1.6 (http://tree.bio.ed.ac.uk/software/tracer/). I used Figtree 1.4.3 (Rambaut
& Drummond, 2010) to illustrate the tree topology.
I visualized the cyt b haplotype distribution by creating a median-joining network using
NETWORK 5.0.0.3. The median-joining method uses a maximum parsimony approach to search
8
for all the shortest phylogenetic trees for a given dataset (Bandelt et al., 1999). I weighted
transversions twice as high as transitions, and used the maximum parsimony option to remove
excessive links and median vectors (Polzin & Daneshmand, 2003). I included all 322 cyt b
sequences of L. albilabris, and 1 cyt b sequence for L. dominicensis in this analysis.
I calculated haplotype diversity (Hd) and nucleotide diversity (πd) for the cyt b data using
ARLEQUIN 3.5.2.2 (Excoffier & Lischer, 2010). I assigned prior population designation based
on the specific locality where the samples were collected on each of the eight islands. I
calculated genetic diversity statistics for all sampling sites represented by at least five specimens.
I inferred gene flow between populations by calculating the fixation index (Fst). Fst values range
from 0 to 1. A value of 0 indicates no population structuring, that is, complete interbreeding
between two populations (panmixia), whereas a value of 1 indicates that all genetic variation is
explained by the population structure, and that the two populations do not share any genetic
diversity. Values reported are means ± 1 SD.
I tested for isolation by distance using the program Alleles in Space (Miller, 2005).
Specifically, the Mantel test option evaluates the correlation between genetic and geographic
distance matrices, using 1,000 permutations and logarithmic transformations. I conducted
analyses for all populations of L. albilabris (PRB and Saint Croix), populations across the PRB,
populations only located in Puerto Rico, and populations only from the Eastern Islands.
Double-digest restriction site associated DNA-sequencing - Laboratory Methods
I selected 119 samples of L. albilabris for double-digest restriction site associated DNA-
sequencing (ddRAD-seq). These samples span the geographic range of the frog in the PRB
(Puerto Rico, n = 70; Vieques, n = 5; Culebra, n = 6; Saint Thomas, n = 7; Saint John, n = 10;
9
Saint Croix, n = 7; Jost Van Dyke, n = 8; and Tortola, n = 6). The ddRAD-seq libraries were
prepared and sequenced at Floragenex Inc. (Portland, OR), as described by Truong et al. (2012),
with the minor adjustments described below. I double-digested 100 – 500 ng of genomic DNA at
37°C using the rare-cutting restriction enzyme Pst1 (5’-CTGCA*G-3’) and the frequent-cutting
restriction enzyme Mse1 (5’-T*TAA-3’). Pst1 P5 adapters containing unique 11 bp sample
identifiers were ligated to individual samples. Next, PCR amplified samples were pooled (5 μl
each) to make the libraries. Fragments were size-selected for the range of 200 – 800 base pairs.
Size-selected samples were amplified in a total reaction volume of 20 μl containing 5 μl of 10-
fold diluted restriction-ligation mixture, 5 ng Illumina P5 primer (5′-
AATGATACGGCGACCACCG-3′), 30 ng Illumina P7 primer (5′-
CAAGCAGAAGACGGCATACGA-3′), 0.2 mM dNTPs, 0.4 U AmpliTaq® (Applied Biosystems),
and 1× AmpliTaq® buffer. The Illumina P7 primer contained an additional +GC to decrease loci
density. Indexed pools were mixed in equimolar ratios and were sequenced together using 101
bp single-end reads on an Illumina HiSeq 2000. The 119 samples were part of two separate
libraries; the first library consisted of 95 samples (Library 1), and the second library consisted of
24 samples (Library 2).
Genomic sequencing methods provide large datasets that may contain artifacts (e.g.
sequencing error, allelic drop out, PCR bias) and uninformative markers that may saturate
relevant population genomic information (Roesti et al., 2012; da Fonesca et al., 2016). Therefore,
I implemented the following bioinformatics protocol to process the data and identify informative
markers. I used STACKS v2.2 to process raw sequence reads (Rochette & Catchen, 2017). I
demultiplexed the raw reads using the PROCESS_RADTAGS command, which identifies sample-
specific barcodes. This command also removes reads with a Phred score < 10 (quality score less
10
than 90%); with default 15% sliding window, as well as ambiguous barcodes. I used the
following series of STACKS commands to assemble and map the demultiplexed sequence data,
and to call single nucleotide polymorphisms (SNPs): the USTACKS command aligned reads,
generated putative de novo loci, and identified SNPs at each locus (Hohenlohe et al., 2010;
Rochette & Catchen, 2017); the CSTACKS command generated a catalog of consensus loci among
all individuals; and the SSTACKS command matched the putative loci of each individual to the
catalog. The POPULATIONS command filtered out SNPs with a maximum observed heterozygosity
of 0.7 and minor allele frequency of 0.05 (Rochette & Catchen, 2017), while focusing on only
single-SNP loci detected in at least two populations and 65% of individuals per population. Next,
I applied the filtering protocol of Puritz et al. (2014). The latter protocol removed individual
samples with more than 40% missing data, as well as genotypes detected in less than half the
total populations (Puritz et al., 2014a, b). I generated a variant calling format (vcf) file containing
16,225 SNPs for analysis. I assessed potential batch affects by assigning the filtered samples to
groups based on the respective sequence library (i.e. Library 1 or Library 2). I removed SNPs
with ≤ 10% representation between the two libraries, and visually inspected the results by using
Principal Component Analysis (PCA) (Luu et al., 2017).
Double-digest restriction site associated DNA-sequencing - Analyses
I calculated observed heterozygosity for each population using the vcf of 16,225 SNPs
and the R package hierfstat (Goudet, 2005). I removed missing data and used the basic.stats
function to calculate observed heterozygosity for each population. I then estimated standard error
in the sciplot R package (Morales, 2017). I assessed population genetic structure within L.
albilabris using the R package LEA (Frichot & François, 2015). This analysis allows for the
11
assessment of population structure among samples, and indicates whether samples represent one
or multiple genetic clusters. Accurate analyses with LEA require missing data to be imputed, or
replaced, with the value “9”. Using a custom script, I further filtered SNPs to only include those
with at least 50% representation among all samples, and imputed the remaining missing data
with the value “9”. I also analyzed population structure with the function snmf. This function
uses a sparse non-negative matrix factorization algorithm to estimate the number of genetic
clusters within the data set, by providing least-squares estimates of ancestry proportions and
calculating an entropy criterion (Frichot et al., 2014). In snmf, the cross-entropy criterion
provides a quality of fit to the statistical model, and can identify the most likely number of
genetic clusters (Frichot et al., 2014). The number of possible genetic cluster(s) was 1 – 10, a
range of values that I predetermined.
12
CHAPTER 3: Results
Bayesian phylogenetic analysis of mitochondrial DNA
Bayesian analysis of mtDNA sequences of L. albilabris revealed a phylogeny with
shallow divergences, and a topology that effectively represents a basal polytomy (Figure 2). I
recovered 28 well-supported (≥ 90% posterior probability values) nodes, or clades. Excluding the
two most inclusive clades, I designated 18 of the remaining 26 well-supported nodes as distinct
phylogroups (PG) (Figure 2). The remainder eight well-supported nodes are nested within five
(i.e. Phylogroups 7, 8, 11, 14, 17) of the 18 phylogroups. The populations and haplotypes that
comprise each phylogroup are listed in Table 2.
Ten of the 18 phylogroups of L. albilabris are comprised of populations exclusively
associated with two of the five physiographic areas of Puerto Rico, the Cordillera Central or the
Lowlands. However, the only phylogroup in the Cordillera Central is Phylogroup 6, which is
made up of a single population from central Puerto Rico (Ciales, Population 11; Figure 2). Of the
18 populations in the Lowlands of Puerto Rico, 14 comprise the remaining nine phylogroups:
Phylogroups 3, 7, 9, 12, 13, 15, 16, 17, 18 (Table 2). There are no well-supported phylogroups
restricted to the Sierra de Cayey, Sierra de Luquillo, or Cuchilla de Pandura physiographic
regions.
I discovered two well-supported nodes in the Eastern Islands, one from Vieques
(Populations 30, 31), and another one from Culebra (Population 32, Phylogroup 1) (Figure 2).
The Vieques clade is nested within a larger phylogroup (Phylogroup 11) that includes
Populations 15, 18–27, and 29 from eastern Puerto Rico (Table 2). The Vieques and Culebra
clades of L. albilabris do not include all the haplotypes identified in each of these islands. One
13
additional haplotype occurs in Vieques (H 64, Population 30); this haplotype does not belong in
any well-supported clade, and is shared with populations from eastern Puerto Rico (Populations
21, 24, 25, 27, 28), Saint John (Population 38), Jost Van Dyke 1, 2 (Populations 39, 40,
respectively), and Tortola 1, 2 (Populations 41, 42, respectively). Culebra harbors three
additional haplotypes (H 45, H 104, H 136) that do not belong within Phylogroup 1. H 45 is
sister to the Culebra clade (Phylogroup 1) (Figure 2), although that node (86% posterior
probability value) did not meet the ≥ 90% threshold to be considered well-supported. H 104 and
H 136 nest within Phylogroup 8, a clade that includes 19 populations (Populations 4–7, 9–13,
16–23, 26–27) across Puerto Rico.
Haplotype network of mitochondrial DNA
The median-joining network for L. albilabris depicts 114 haplotypes private to Puerto
Rico (Figure 3). The most frequent haplotype (H 115) was detected in 30 individuals from 12
different populations across Puerto Rico. Each of the Eastern Islands harbors at least one private
haplotype (Table 3; Figure 3). Six haplotypes (H 3, H 4, H 64, H 97, H 98, H 99) are private to
Vieques 1, 2 (Populations 30 and 31, respectively), and five (H 3, H 4, H 97, H 98, H 99) of
them form a cluster separated by 1–4 mutational steps. I detected five private haplotypes on
Culebra (Population 32), where three haplotypes (H 43, H 45, H 60) form a cluster separated by
1–3 mutations. Saint Thomas 2 (Population 34, H 8) and Saint Croix 2 (Population 36, H 30)
each harbors a single private haplotype. Saint John (Population 38) has two private haplotypes
(H 5, H 109), whereas Jost Van Dyke 1 (Population 39, H 111) and Tortola 2 (Population 42, H
107) each has a single private haplotype.
14
Network analysis detected four haplotypes present on more than one island (Table 3;
Figure 3). Haplotype 6 occurs on Puerto Rico, Vieques, Saint John, Jost Van Dyke, and Tortola.
Haplotype 10 occurs on Saint Thomas and Saint Croix. Haplotypes 28, H 108, and H 121 are
recognized as equivalent in NETWORK because they differ by three unspecified base pairs, and
they occur on Saint Thomas, Tortola, and Puerto Rico (Río Grande, Population 28), respectively.
Leptodactylus dominicensis (Hispaniola) and two samples from northeastern Puerto Rico
(Dorado, Population 14) share H 51.
Haplotype diversity and nucleotide diversity of mitochondrial DNA
I calculated haplotype diversity (Hd) for all populations represented by 5 individuals
(Table 4). Hd is highly variable for L. albilabris populations across the PRB and Saint Croix
(average Hd = 0.81, range = 0.0 – 1.0; Table 4). The populations with the greatest haplotype
diversity (Hd = 1.0) occur in eastern Puerto Rico: Aguas Buenas (Population 20), Cayey 1
(Population 21), Cayey 2 (Population 22), and Juncos (Population 27). The Saint Croix
populations 1, 2 (Populations 35, 36, respectively) lack haplotype diversity (Hd = 0.0).
Within Puerto Rico, average Hd = 0.89 (range = 0.25 – 1.0). As previously stated, four
populations from Puerto Rico displayed the highest haplotype diversity, whereas Peñuelas
(Population 9) had the lowest Hd value. The Eastern Island populations of L. albilabris had lower
Hd estimates (average Hd = 0.63, range = 0.25 – 0.89) than those from Puerto Rico. Vieques 1
(Population 30) had the highest Hd, whereas Jost Van Dyke 1 (Population 39) had the lowest one.
I also calculated nucleotide diversity (πd) for all populations represented by 5
individuals (Table 4). πd among populations of L. albilabris across the PRB and Saint Croix
averaged 0.0059 (range = 0.0 – 0.0121). The greatest πd estimate corresponded to Aguas Buenas
15
(Population 20, Puerto Rico), whereas the smallest πd values occur in Saint Thomas 1
(Population 33, πd = 0.0), and Saint Croix 1, 2 (Population 35, πd = 0.0, and Population 36, πd =
0.0002).
Within Puerto Rico, average πd = 0.007 (range = 0.0006 – 0.0121). As I mentioned,
Aguas Buenas (Population 20) had the highest πd estimate, whereas Peñuelas (Population 9) had
the lowest one. The πd estimates among the Eastern Islands populations of L. albilabris averaged
0.0032 (range = 0.0 – 0.0097), a value noticeably smaller than that for the Puerto Rican
populations. In the Eastern Islands, Culebra (Population 32) had the greatest πd estimate, whereas
Saint Thomas 1 (Population 33) had the smallest one, as reported above.
Population structuring and association between geographic and genetic distance calculated
using mitochondrial DNA
Fixation index (Fst) estimates for populations of L. albilabris across the PRB and Saint
Croix averaged 0.15 (range = 0.0 – 0.84; Table 5). Two localities from Saint Croix, Population
35 (average Fst = 0.46, range = -0.09 – 0.84) and Population 36 (average Fst = 0.48, range = -0.09
– 0.79) had the greatest overall Fst estimates. Interestingly, these Saint Croix localities do not
exhibit genetic structuring between them (Fst = 0.0). The Puerto Rican locality of Juncos
(Population 27) exhibited the lowest average Fst value (0.03, range -0.04 – 0.42). The Mantel test
indicated no correlation between genetic and geographic distance among all (PRB and Saint
Croix) populations of L. albilabris (r = 0.01; P = 0.34), or among populations from the PRB (r =
0.02; P = 0.19).
Puerto Rican populations of L. albilabris exhibited relatively low population genetic
structuring (average Fst = 0.08, range = 0.0 – 0.48). Peñuelas (Population 9) displayed the highest
16
values (average Fst = 0.35, range = 0.12 – 0.48), and Juncos (Population 27) had the lowest
estimates, as reported above. The Mantel test did not detect a correlation between genetic and
geographic distance within Puerto Rico (r = 0.01; P = 0.28).
The Fst estimates for L. albilabris populations in the Eastern Islands (average = 0.35,
range = 0.0 – 0.84) indicate higher levels of population genetic structuring than among Puerto
Rican populations of this frog. Jost Van Dyke 1 (Population 39) exhibited the highest average Fst
value (0.47, range = 0.0 – 0.84). On the contrary, and interestingly, this Jost Van Dyke
population and Tortola 2 (Population 42) exhibited zero genetic structuring between them (Fst =
0.0). Except for the aforementioned Jost Van Dyke 1 and Tortola 2 populations, Vieques 1
(Population 30) displayed the smallest Fst values (average = 0.28, range = 0.12 – 0.48). The
Mantel test indicates a correlation between genetic and geographic distance among L. albilabris
populations in the Eastern Islands (r = 0.47; P = 0.001).
Double-digest restriction site associated DNA-sequencing SNP-filtering
I recovered 341,505,569 reads from the first library, and 313,493,233 reads from the
second library, for a total of 654,998,802 reads from the 119 samples. Read depth for all samples
averaged 26.52 reads per sample (range = 16.86 – 45.22 reads). Prior to filtering, my
bioinformatics protocol generated a catalog of 1,570,383 genotyped loci and 2,497,178 SNPs.
After my filtering procedure I retained 16,225 single-SNP loci from 114 samples in the final vcf
file. My bioinformatics protocol detected five samples that did not pass the filtering criteria, and
I removed those samples from the dataset.
17
The filtering protocol to assess batch effects resulted in 4,046 SNPs. I visually inspected
the results through PCA, and the patterns were largely unchanged. Therefore, I determined that
my filtering protocol was thorough, and I conducted further analyses using all 16,225 SNPs.
Population genomic analyses – heterozygosity and phylogenomic structure
Heterozygosity estimates among all populations of L. albilabris (PRB and Saint Croix)
were low (average = 0.17, range = 0.10 – 0.23; Table 6). Aguas Buenas (Population 20, Puerto
Rico) and Saint Croix 2 (Population 36) exhibited the greatest and smallest heterozygosity
estimates, respectively.
Within Puerto Rico, heterozygosity estimates showed little variation (average = 0.19,
range 0.17 – 0.23; Table 6). Aguas Buenas (Population 20) and Moca (Population 2) displayed
the highest and lowest estimates, respectively. Heterozygosity estimates of the Eastern Islands
populations of L. albilabris (average = 0.12, range = 0.10 – 0.17) were lower than those of
Puerto Rican populations. Vieques 1 (Population 30) and Jost Van Dyke 1 (Population 39)
displayed the greatest and smallest heterozygosity values, respectively.
After filtering for missing data and conducting imputation, I generated a dataset with
16,140 SNPs representing 114 individuals. Clustering analyses using snmf identified five L.
albilabris genetic clusters with little admixture (Figure 4). Two genetic clusters were detected on
Puerto Rico, although only the first cluster was exclusive to this island. The first genetic cluster
(Cluster 1) consisted of three of the six populations from the Cordillera Central, specifically
Yauco (Population 6), Juana Díaz (Population 10), and Ciales (Population 11). The second
genetic cluster (Cluster 2) was formed by all other populations from the remaining four
physiographic regions of Puerto Rico (i.e. Sierra de Cayey, Sierra de Luquillo, Cuchilla de
18
Pandura, Lowlands), and the neighboring islands of Vieques 1 (Population 30) and Culebra
(Population 32). The third genetic cluster (Cluster 3) comprised populations from Saint Thomas
1 (Population 33) and Saint Croix 2 (Population 36). The fourth genetic cluster (Cluster 4)
corresponded to Saint John (Population 38), the only island that comprised an independent,
distinct genetic cluster. The fifth genetic cluster (Cluster 5) was formed by populations from Jost
Van Dyke 1, 2 (Populations 39, 40) and Tortola (Population 42).
Genetic clustering analysis revealed incongruences of cluster membership at the
population level. I detected two instances in which populations had individuals that belong to
different genetic clusters. (i) In Saint John (Population 38), nine individuals belong to Cluster 4,
and one belongs in Cluster 5. (ii) In Jost Van Dyke 1 (Population 39), one individual belongs to
Cluster 4, and the remaining six belong in Cluster 5.
19
CHAPTER 4: Discussion
Phylogeographic and population genetic studies can be more informative when the spatial
and temporal history of the study system are taken into consideration. The sea level changes of
Pleistocene glacial-interglacial cycles are associated with distributional shifts of taxa on a global
basis (Hewitt, 2000; Vasconcellos et al., 2019). Lower sea levels during glacial periods revealed
land bridges in island systems, providing opportunities for organisms to migrate between
formerly separated islands (Garg et al., 2018; Weigelt et al., 2016). On the other hand, higher sea
levels during interglacial periods submerged land bridges and fragmented islands (Weigelt et al.,
2016), potentially isolating previously connected populations. Repeated episodes of island
connectivity and fragmentation constitute a dynamic scenario that may have influenced the
evolutionary history of organisms native to island systems (Potter et al., 2018; Parvizi et al.,
2018; Sánchez-Montes et al., 2019). The Puerto Rican Bank (PRB) is an archipelago in the
eastern Caribbean Sea that exhibited periodic connectivity and fragmentation during the
Pleistocene (Heatwole & MacKenzie, 1967; Hedges, 1999; Renken et al., 2002). I herein
conducted a genetic assessment of populations of Leptodactylus albilabris across the PRB and
Saint Croix, to determine the potential effect of changes in island configuration on the
demographic history of this frog.
The effect of Puerto Rico’s physiography and Pleistocene climate on patterns of genetic
variation in Leptodactylus albilabris
I hypothesized that the boundaries of the five physiographic regions of Puerto Rico
(Cordillera Central, Sierra de Cayey, Sierra de Luquillo, Cuchilla de Pandura, Lowlands)
facilitate geographic structuring among populations of L. albilabris from those regions. The
20
results from the mtDNA and ddRAD-seq analyses only provided partial, incongruent support for
this hypothesis. Phylogenetic analysis of cytochrome b haplotypes detected well-supported
phylogroups associated with the Cordillera Central and the Lowlands (Figure 2). However, the
only phylogroup in the Cordillera Central compromises a single population from central Puerto
Rico, implying that the vast majority of populations from the Cordillera Central do not form a
distinct genetic cluster. Conversely, populations from the Lowlands formed nine different
phylogroups, reflecting a higher degree of structuring of populations of L. albilabris from low
elevation areas around Puerto Rico. The observation that several mtDNA haplotypes occur in
more than one physiographic region, coupled with the low Fst values among Puerto Rican
populations, suggest relatively unrestricted, recent or ongoing gene flow among L. albilabris
from much of the island.
The findings from the ddRAD-seq clustering analyses coincided in part with those from
the mitochondrial sequences. Specifically, genetic clustering of the much larger genomic dataset
also identified two genetic groups in Puerto Rico. Cluster 1 comprises three populations from the
Cordillera Central (Yauco, Population 6; Juana Díaz, Population 10; Ciales, Population 11)
(Figure 4), reminiscent of the mtDNA-inferred Phylogroup 6. However, in contrast to the
mtDNA dataset, the second genetic group (Cluster 2) identified by the genomic data is made up
of populations from the four remainder regions of Puerto Rico (Sierra de Cayey, Sierra de
Luquillo, Cuchilla de Pandura, Lowlands), and the neighboring islands of Vieques and Culebra.
Individuals collected in Juana Díaz, southcentral Puerto Rico, exemplify the distinction between
the two clusters. The four samples collected in the northern, mountainous section of this
municipality (Population 10) belong to Cluster 1, whereas the individual collected in the coastal
Lowlands in southern Juana Díaz (Population 12) belongs in Cluster 2. Interestingly, L.
21
albilabris from higher elevations are larger than those from lower elevations (Heatwole et al.,
1968), and coastal individuals exhibit a higher salinity tolerance (Gómez-Mestre & Tejedo,
2003). These biological differences may facilitate genetic divergence between L. albilabris
populations from the Cordillera Central and those from other regions of Puerto Rico. In regards
to L. albilabris from Vieques and Culebra, receding coastlines of these two islands during glacial
periods likely allowed coastal populations in eastern Puerto Rico to expand into these adjacent
islands (Figure 5D).
Collectively, analyses of the mtDNA and ddRAD-seq datasets revealed that although
some degree of genetic diversification has occurred, most Puerto Rican populations of L.
albilabris are genetically homogeneous. This finding is not intuitive, given that the occurrence of
L. albilabris depends on at least temporal availability of freshwater, because these frogs have a
free-living aquatic tadpole (Dent, 1956; Heatwole et al., 1968). Moisture and precipitation are
not evenly distributed throughout Puerto Rico; regional habitat characteristics of the island range
from higher elevation wet forests in the Cordillera Central, Sierra de Cayey and Sierra de
Luquillo, to lower elevation moist areas in the Cuchilla de Pandura, and to low elevation
subtropical dry forests predominantly located in the Lowlands of southern Puerto Rico, in the
rain shadow of the Cordillera Central (Ewel & Whitmore, 1973; Helmer et al., 2002; Renken et
al., 2002). (Subtropical dry forests also occur on the northeast coast of Puerto Rico, and on
Vieques, Culebra, and the United States and British Virgin Islands [Murphy et al., 1995].) Due to
its reproductive mode, arid conditions are expected to limit the distribution and mobility of L.
albilabris, and thus represent barriers to gene flow that will facilitate genetic divergence among
populations. These circumstances would have been especially true during the late Pleistocene
(ca. 95 – 10 kya), because the Caribbean was then cooler, but drier than at the present time
22
(Royer et al., 2017; Warken et al., 2019). However, L. albilabris tadpoles have higher heat
tolerance, which suggests that they are adapted to breeding in ephemeral and/or shallow bodies
of water (Heatwole et al., 1968). In fact, in Jost Van Dyke (Population 39) I collected specimens
in a small ditch next to an unpaved road. The larval stage of L. albilabris is relatively short,
tadpoles can metamorphose within 21–35 days after hatching (Dent, 1956; Joglar 2005; Flores-
Nieves et al., 2014). Additionally, L. albilabris at various stages of metamorphosis can be found
in muddy areas or under debris when standing water is no longer present (Heatwole et al., 1968).
These physiological traits limit the frogs’ dependence on freshwater, and thus increase
population connectivity and gene flow throughout much of Puerto Rico.
Isolation by distance of Leptodactylus albilabris populations across the Puerto Rican Bank
I hypothesized that populations of L. albilabris exhibit “isolation by distance” (IBD)
across the PRB. That is, I expected populations from Puerto Rico to be most divergent from the
populations from the eastern most British Virgin Islands (i.e. Jost Van Dyke and Tortola).
Mantel tests of the mtDNA dataset conducted at different spatial scales did not detect any
correlations between genetic and geographic distance among populations of L. albilabris within
Puerto Rico, across the PRB, or among all populations across the PRB and Saint Croix. Failure
to detect a signal for IBD suggests that gene flow occurred across wide spatial scales during
periods of connectivity in the PRB, or that the White-lipped frog experienced recent population
(spatial) expansion, perhaps facilitated by increasing land area during glacial periods (Reynolds
et al., 2017). Anolis cristatellus (Puerto Rican crested anole) has a similar distribution to L.
albilabris, and Eastern Islands populations of this lizard do not exhibit IBD either (Reynolds et
al., 2017).
23
On the contrary, Mantel tests restricted to the Eastern Islands populations of L. albilabris
detected IBD among the localities. Additionally, these populations displayed relatively high Fst
values. These findings suggest limited gene flow among Eastern Islands L. albilabris, and
possible genetic drift, because genetic drift is a phenomenon typically associated with small,
isolated populations (Wright, 1931; Nei et al., 1975; Frankham, 1997). The divergence among
the Eastern Islands populations of L. albilabris may be attributed to isolation caused by island
fragmentation during interglacial periods, and/or to limited gene flow even during periods of
island connectivity. As I mentioned, arid climate and xeric habitats characterized the PRB during
the Pleistocene (Pregill & Olson, 1981; Royer et al., 2017; Warken et al., 2019). Therefore, the
frog’s ability to move among localities corresponding to the present-day Eastern Islands may
have been limited by scarcity of suitable habitat on the land bridges. Ecological barriers to gene
flow, rather than geographic distance, may then explain genetic divergence among Eastern
Islands L. albilabris.
Ancestral area of Leptodactylus albilabris
As I stated, L. albilabris has a widespread distribution across the PRB, particularly in
Puerto Rico, where the species occurs in natural, residential, and commercial areas. I proposed
that L. albilabris evolved in Puerto Rico, the largest island of the PRB. Biogeographic theory
posits that greater genetic diversity is indicative of a longer demographic history; that is, older
populations have had more time to evolve (Taberlet et al., 2002; Provan & Bennett, 2008) and
accumulate genetic differences. Genetic diversity estimates calculated from mtDNA and
ddRAD-seq support the prediction that Puerto Rico is the ancestral area of L. albilabris, because
the Puerto Rican populations of this frog exhibit greater genetic diversity than those from the
24
Eastern Islands and Saint Croix (Tables 4, 6). Further, more private mtDNA haplotypes were
detected in Puerto Rico than in the Eastern Islands (Figure 3). Eleutherodactylus antillensis
(Red-eyed Coquí) is a frog with a similar distribution to that of L. albilabris, and genetic
analyses also suggest that E. antillensis evolved in Puerto Rico, and colonized the Eastern
Islands from sources in eastern Puerto Rico via land bridges during island connectivity (Barker et
al., 2012).
Nevertheless, the lower genetic diversity of Eastern Islands L. albilabris may be a
consequence of island area. Island biogeographic theory asserts that larger islands harbor more
ecological diversity, whether it is due to total physical area or to habitat heterogeneity
(MacArthur & Wilson, 1967; Ricklefs & Lovett, 1999). It is thus possible that the greater levels
of genetic diversity in Puerto Rican L. albilabris may simply result from the greater land area of
Puerto Rico, and that the lower genetic diversity in Eastern Islands L. albilabris is a demographic
consequence of island fragmentation (Frankham, 1997; White & Searle, 2007; Furlan et al.,
2012; Wang et al., 2014) in the PRB. Specifically, range contraction and population isolation by
rising sea levels could have led to population bottleneck and genetic drift (Hoffman & Blouin,
2004; Wang et al., 2014), causing a reduction in the genetic diversity of Eastern Islands
populations (Table 4, 6). Phylogenetic analysis of unique mtDNA haplotypes recovered a
topology that effectively represents a basal polytomy (Figure 2), and therefore is uninformative
regarding the question of the geographic origin of L. albilabris. All these scenarios allow for the
possibility that the White-lipped frog may have originated in the Eastern Islands, and then
colonized and differentiated genetically in Puerto Rico. In conclusion, the ancestral area of L.
albilabris remains equivocal.
25
Origins of the Saint Croix populations of Leptodactylus albilabris
The origin of the Saint Croix populations of L. albilabris is unclear, given that this island
has not had a land connection to the PRB (Gill et al., 1989). Ocean currents around the PRB
generally flow in a northwestern direction (Heatwole & Mackenzie, 1967), suggesting that L.
albilabris could have evolved in situ on Saint Croix and later colonized the PRB. Typically,
ancestral populations exhibit relatively high levels of genetic diversity and population structuring
(Reilly et al., 2019), but Saint Croix L. albilabris do not display these patterns. I sampled three
localities across the island (Figure 1), and detected only two haplotypes among 18 individuals:
one private mtDNA haplotype in one frog, and another haplotype shared by the remaining 17
animals from Saint Croix. The latter haplotype is also shared by seven individuals from Saint
Thomas (Figure 3; Table 3). Clustering analyses of the genomic data confirmed the genetic
similarity between Saint Thomas (Population 33) and Saint Croix (Populations 36) L. albilabris,
and depicted the two populations as a single cluster with no admixture (Figure 4). The low
genetic diversity in populations of L. albilabris in Saint Croix suggests two possible scenarios:
the species experienced a selective sweep across the island (Tajima, 1989; Nguiffo et al., 2019),
or it arrived on Saint Croix relatively recently, possibly from a source population in Saint
Thomas. The selective sweep scenario in turn implies that the Saint Thomas and Saint Croix
populations evolved the same cytochrome b haplotype through convergence, perhaps due to a
similar selective pressure (Losos, 2011; Messer & Petrov, 2013). The latter evolutionary vista
may not represent the more parsimonious explanation to account for the presence of L. albilabris
on Saint Croix.
Perhaps a more likely scenario is that L. albilabris originated in the PRB, and
subsequently arrived on Saint Croix by either natural means or through human-mediated
26
introduction(s). Natural immigration requires L. albilabris to have rafted to Saint Croix. As
mentioned before, the surface currents in the eastern Caribbean Sea generally have a
northwestern direction (Heatwole & Mackenzie, 1967), making rafting southward from a source
in the PRB to Saint Croix an improbable event. Nevertheless, ocean dynamics can be impacted
during tropical storms and hurricanes (Ezer, 2019), and thus surface currents could temporarily
change course and cause flotsam to drift in atypical directions. Even when rafting occurs, the
survival of rafting organisms in the PRB is low (Heatwole & Levins, 1972; Ricklefs &
Bermingham, 2008). Salt water is physiologically stressful for anurans (Balinsky, 1981;
Duellman & Trueb, 1994), and the probability of oceanic dispersal of different frog species can
vary, because of differences in physiological tolerance to salinity and habitat requirements
(Duryea et al., 2015). Yet, oceanic dispersal by anurans has been documented (e.g. Hedges et al.,
1992; Vences et al., 2003; Heinicke et al., 2007; Bell et al., 2015), and there are published
accounts of oceanic dispersal by nonavian reptiles in the Caribbean Sea (e.g. Corallus
hortulanus, Garden tree boa, Henderson & Hedges, 1995; Green iguana, Iguana iguana, Censky
et al., 1998; Anolis sagrei, Brown anole, Calsbeek & Smith, 2003). Accidental or even deliberate
introduction of L. albilabris to Saint Croix is also possible. Maritime travel by humans has had a
noticeable impact on biodiversity globally (Reilly et al. 2019), including the transportation of
frog species among Caribbean islands (MacLean, 1982; Kaiser et al., 2002; Rödder, 2009;
Barker et al., 2012; Barker & Rodríguez-Robles, 2017). For example, E. antillensis and
Osteopilus septentrionalis (Cuban Tree frog) have been documented ‘hitchhiking’ on
landscaping materials that were being transported between islands of the PRB and Saint Croix
(Townsend et al., 2000; Powell et al., 2005; Powell, 2006; Perry et al., 2006; Platenberg, 2007;
Powell et al., 2011).
27
Systematic status of Leptodactylus dominicensis
Phenotypical, behavioral, and genetic data clearly indicate that Leptodactylus
dominicensis (Cochran, 1823) is closely related to L. albilabris (Gunther, 1859). In fact, several
authors have placed L. dominicensis in the synonymy of L. albilabris (Heyer, 1978; Hedges &
Heinicke, 2007; de Sá et al., 2008). I conducted the most inclusive genetic assessment of L.
albilabris throughout its range to date, and relied on this sampling to assess the taxonomic
position of L. dominicensis. Phylogenetic analysis conclusively demonstrated that the lone
available cytochrome b sequence of L. dominicensis nests within those of L. albilabris. Indeed,
two individuals of L. albilabris from Puerto Rico’s northern coast (Dorado, Population 14) share
the same mtDNA haplotype (H 51) with L. dominicensis (Figure 2). My findings thus support the
conclusion that L. dominicensis should not be considered a separate species, a taxonomic
arrangement that I follow hereafter.
Leptodactylus albilabris is known only from a small area in northeastern Hispaniola.
Individuals of this frog have been collected from El Seibo Province, Dominican Republic
(Cochran, 1941; Heyer, 1978; Hedges & Heinicke, 2007), which is adjacent to Samaná Bay. As
with L. albilabris in Saint Croix, the presence of L. albilabris on Hispaniola likely represents a
relatively recent colonization event, by natural or anthropogenic means. Oceanic currents in the
eastern Caribbean Sea could have transported L. albilabris via flotsam west into Samaná Bay.
Accidental or deliberate introduction of L. albilabris to Hispaniola may also have occurred, as
human-mediated introduction of Eleutherodactylus johnstonei, Lesser Antillean Frog (Barbour,
1930; Censky, 1989; Lever, 2003; Rödder, 2009), and Osteopilus septentrionalis, Cuban
Treefrog (Townsend et al., 2000; Powell et al., 2005; Powell, 2006; Perry et al., 2006), to various
islands in the Caribbean Sea has been documented.
28
Concluding remarks
Elucidating the genetic architecture of L. albilabris across the PRB illustrated the
importance of integrating geologic information into studies of the evolutionary history of a
species. I used molecular data to evaluate how eustatic sea level changes during the Pleistocene
Epoch may have shaped the genetic profile of this frog. I detected interesting genetic patterns in
L. albilabris, both within Puerto Rico and throughout the species’ range in the PRB. The
ddRAD-seq clustering analyses uncovered a divergence between L. albilabris populations from
Puerto Rico’s Cordillera Central and the rest of the island. Future studies should investigate
which biological and/or environmental factor(s) may facilitate this differentiation, and assess
whether traits other than body size and salinity tolerance vary between populations from higher
and lower elevation areas in Puerto Rico. Additionally, little is known about the dispersal ability
of L. albilabris. It will therefore be informative to conduct radiotracking and mark-recapture
studies to characterize the spatial ecology (e.g. movement patterns, home range size), and by
extension, the degree of connectivity among populations of the species. As a final point, my
study contributed to our increasing understanding of the biogeography of the Puerto Rican Bank
(e.g. Barker et al., 2012; Falk & Perkins, 2013; Papadopoulou & Knowles, 2015, 2017;
Rodríguez-Robles et al., 2015; Reynolds et al., 2017).
29
Tab
le 1
. Spe
cies
, pop
ulat
ion
num
ber,
field
num
ber,
coun
try, s
tate
, isl
and,
mun
icip
ality
, lat
itude
(h =
deg
rees
, min
= m
inut
es),
and
long
itude
of t
he lo
calit
ies o
f the
spec
imen
s of L
epto
dact
ylus
poe
cilo
chilu
s, L.
dom
inic
ensi
s, an
d L.
alb
ilabr
is u
sed
in th
is st
udy.
For
L.
poec
iloch
ilus,
the
mus
eum
cat
alog
num
ber i
s in
pare
nthe
ses;
for L
. dom
inic
ensi
s, th
e G
enB
ank
acce
ssio
n nu
mbe
r is i
n pa
rent
hese
s.
BB
= B
ritta
ny B
arke
r Fie
ld S
erie
s, B
VI =
Brit
ish
Virg
in Is
land
s, C
U =
Cul
ebra
, JA
R =
Javi
er A
. Rod
rígue
z Fi
eld
Serie
s, JV
D =
Jos
t
Van
Dyk
e, P
R =
Pue
rto R
ico,
SB
H =
S. B
lair
Hed
ges F
ield
Ser
ies,
STC
= S
aint
Cro
ix, S
TJ =
Sai
nt Jo
hn, S
TT =
Sai
nt T
hom
as, T
O =
Torto
la, U
K =
Uni
ted
Kin
gdom
, US
= U
nite
d St
ates
of A
mer
ica,
USN
M =
Nat
iona
l Mus
eum
of N
atur
al H
isto
ry, S
mith
soni
an
Inst
itutio
n, W
ashi
ngto
n, D
.C.,
USV
I = U
nite
d St
ates
Virg
in Is
land
s, V
I = V
iequ
es. S
ampl
es o
f L. a
lbila
bris
are
list
ed in
ord
er b
y
popu
latio
n nu
mbe
r, fr
om w
est t
o ea
st (F
igur
e 1)
.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
O
utgr
oup
Le
ptod
acty
lus
poec
iloch
ilus
(USN
M 5
6502
9)
—
—
Pana
má
—
—
Pana
má
Prov
ince
—
—
—
—
30
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
In
grou
p
Lept
odac
tylu
s do
min
icen
sis
(EF0
9139
3)
—
SBH
192
453
D
omin
ican
R
epub
lic
H
ispa
niol
a El
Sei
bo
Prov
ince
—
—
—
—
Le
ptod
acty
lus
albi
labr
is
1 JA
R 2
905
US
PR
PR
Cab
o R
ojo
18°
01.0
30' N
67
° 08
.955
' W
Lept
odac
tylu
s al
bila
bris
1
JAR
290
6 U
S PR
PR
C
abo
Roj
o 18
° 01
.030
' N
67°
08.9
55' W
Lept
odac
tylu
s al
bila
bris
1
JAR
290
7 U
S PR
PR
C
abo
Roj
o 18
° 01
.030
' N
67°
08.9
55' W
Lept
odac
tylu
s al
bila
bris
1
JAR
290
8 U
S PR
PR
C
abo
Roj
o 18
° 01
.030
' N
67°
08.9
55' W
Lept
odac
tylu
s al
bila
bris
1
JAR
290
9 U
S PR
PR
C
abo
Roj
o 18
° 01
.030
' N
67°
08.9
55' W
Lept
odac
tylu
s al
bila
bris
1
JAR
291
0 U
S PR
PR
C
abo
Roj
o 18
° 01
.030
' N
67°
08.9
55' W
Lept
odac
tylu
s al
bila
bris
1
JAR
291
1 U
S PR
PR
C
abo
Roj
o 18
° 01
.030
' N
67°
08.9
55' W
Lept
odac
tylu
s al
bila
bris
1
JAR
291
2 U
S PR
PR
C
abo
Roj
o 18
° 01
.030
' N
67°
08.9
55' W
Lept
odac
tylu
s al
bila
bris
1
JAR
291
3 U
S PR
PR
C
abo
Roj
o 18
° 01
.030
' N
67°
08.9
55' W
Lept
odac
tylu
s al
bila
bris
1
JAR
291
4 U
S PR
PR
C
abo
Roj
o 18
° 01
.030
' N
67°
08.9
55' W
31
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
2 JA
R 2
740
US
PR
PR
Moc
a 18
° 23
.477
' N
67°
06.0
78' W
Lept
odac
tylu
s al
bila
bris
2
JAR
274
1 U
S PR
PR
M
oca
18°
23.4
77' N
67
° 06
.078
' W
Lept
odac
tylu
s al
bila
bris
2
JAR
274
2 U
S PR
PR
M
oca
18°
23.4
77' N
67
° 06
.078
' W
Lept
odac
tylu
s al
bila
bris
2
JAR
274
3 U
S PR
PR
M
oca
18°
23.4
77' N
67
° 06
.078
' W
Lept
odac
tylu
s al
bila
bris
2
JAR
274
4 U
S PR
PR
M
oca
18°
23.4
77' N
67
° 06
.078
' W
Lept
odac
tylu
s al
bila
bris
2
JAR
274
5 U
S PR
PR
M
oca
18°
23.4
77' N
67
° 06
.078
' W
Lept
odac
tylu
s al
bila
bris
2
JAR
274
7 U
S PR
PR
M
oca
18°
23.4
77' N
67
° 06
.078
' W
Lept
odac
tylu
s al
bila
bris
2
JAR
274
8 U
S PR
PR
M
oca
18°
23.4
77' N
67
° 06
.078
' W
Lept
odac
tylu
s al
bila
bris
3
JAR
262
5 U
S PR
PR
Is
abel
a 18
° 30
.636
' N
67°
05.7
28' W
Lept
odac
tylu
s al
bila
bris
3
JAR
262
6 U
S PR
PR
Is
abel
a 18
° 30
.636
' N
67°
05.7
28' W
Lept
odac
tylu
s al
bila
bris
3
JAR
262
7 U
S PR
PR
Is
abel
a 18
° 30
.636
' N
67°
05.7
28' W
Lept
odac
tylu
s al
bila
bris
3
JAR
262
8 U
S PR
PR
Is
abel
a 18
° 30
.636
' N
67°
05.7
28' W
Lept
odac
tylu
s al
bila
bris
3
JAR
262
9 U
S PR
PR
Is
abel
a 18
° 30
.636
' N
67°
05.7
28' W
Lept
odac
tylu
s al
bila
bris
3
JAR
263
0 U
S PR
PR
Is
abel
a 18
° 30
.636
' N
67°
05.7
28' W
32
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
3 JA
R 2
631
US
PR
PR
Isab
ela
18°
30.6
36' N
67
° 05
.728
' W
Lept
odac
tylu
s al
bila
bris
3
JAR
263
2 U
S PR
PR
Is
abel
a 18
° 30
.636
' N
67°
05.7
28' W
Lept
odac
tylu
s al
bila
bris
3
JAR
263
3 U
S PR
PR
Is
abel
a 18
° 30
.636
' N
67°
05.7
28' W
Lept
odac
tylu
s al
bila
bris
3
JAR
263
4 U
S PR
PR
Is
abel
a 18
° 30
.636
' N
67°
05.7
28' W
Lept
odac
tylu
s al
bila
bris
4
JAR
307
0 U
S PR
PR
M
ayag
üez
18°
11.3
03' N
67
° 03
.626
' W
Lept
odac
tylu
s al
bila
bris
4
JAR
307
2 U
S PR
PR
M
ayag
üez
18°
11.3
03' N
67
° 03
.626
' W
Lept
odac
tylu
s al
bila
bris
4
JAR
307
4 U
S PR
PR
M
ayag
üez
18°
11.3
03' N
67
° 03
.626
' W
Lept
odac
tylu
s al
bila
bris
4
JAR
307
5 U
S PR
PR
M
ayag
üez
18°
11.3
03' N
67
° 03
.626
' W
Lept
odac
tylu
s al
bila
bris
4
JAR
307
6 U
S PR
PR
M
ayag
üez
18°
11.3
03' N
67
° 03
.626
' W
Lept
odac
tylu
s al
bila
bris
5
JAR
289
3 U
S PR
PR
G
uáni
ca
17°
58.7
51' N
66
° 57
.089
' W
Lept
odac
tylu
s al
bila
bris
5
JAR
289
4 U
S PR
PR
G
uáni
ca
17°
58.7
51' N
66
° 57
.089
' W
Lept
odac
tylu
s al
bila
bris
5
JAR
289
5 U
S PR
PR
G
uáni
ca
17°
58.7
51' N
66
° 57
.089
' W
Lept
odac
tylu
s al
bila
bris
5
JAR
289
6 U
S PR
PR
G
uáni
ca
17°
58.7
51' N
66
° 57
.089
' W
Lept
odac
tylu
s al
bila
bris
5
JAR
289
7 U
S PR
PR
G
uáni
ca
17°
58.7
51' N
66
° 57
.089
' W
33
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
5 JA
R 2
898
US
PR
PR
Guá
nica
17
° 58
.751
' N
66°
57.0
89' W
Lept
odac
tylu
s al
bila
bris
5
JAR
289
9 U
S PR
PR
G
uáni
ca
17°
58.7
51' N
66
° 57
.089
' W
Lept
odac
tylu
s al
bila
bris
5
JAR
290
0 U
S PR
PR
G
uáni
ca
17°
58.7
51' N
66
° 57
.089
' W
Lept
odac
tylu
s al
bila
bris
5
JAR
290
1 U
S PR
PR
G
uáni
ca
17°
58.7
51' N
66
° 57
.089
' W
Lept
odac
tylu
s al
bila
bris
5
JAR
290
2 U
S PR
PR
G
uáni
ca
17°
58.7
51' N
66
° 57
.089
' W
Lept
odac
tylu
s al
bila
bris
6
JAR
307
7 U
S PR
PR
Y
auco
18
° 09
.100
' N
66°
53.0
99' W
Lept
odac
tylu
s al
bila
bri s
6
JAR
307
8 U
S PR
PR
Y
auco
18
° 09
.100
' N
66°
53.0
99' W
Lept
odac
tylu
s al
bila
bris
6
JAR
307
9 U
S PR
PR
Y
auco
18
° 09
.100
' N
66°
53.0
99' W
Lept
odac
tylu
s al
bila
bris
6
JAR
308
0 U
S PR
PR
Y
auco
18
° 09
.100
' N
66°
53.0
99' W
Lept
odac
tylu
s al
bila
bris
6
JAR
308
1 U
S PR
PR
Y
auco
18
° 09
.100
' N
66°
53.0
99' W
Lept
odac
tylu
s al
bila
bris
6
JAR
308
3 U
S PR
PR
Y
auco
18
° 09
.100
' N
66°
53.0
99' W
Lept
odac
tylu
s al
bila
bris
6
JAR
308
4 U
S PR
PR
Y
auco
18
° 09
.100
' N
66°
53.0
99' W
Lept
odac
tylu
s al
bila
bris
6
JAR
308
5 U
S PR
PR
Y
auco
18
° 09
.100
' N
66°
53.0
99' W
Lept
odac
tylu
s al
bila
bris
6
JAR
308
6 U
S PR
PR
Y
auco
18
° 09
.100
' N
66°
53.0
99' W
34
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
6 JA
R 3
087
US
PR
PR
Yau
co
18°
09.1
00' N
66
° 53
.099
' W
Lept
odac
tylu
s al
bila
bris
7
JAR
285
4 U
S PR
PR
H
atill
o 18
° 29
.175
' N
66°
49.9
12' W
Lept
odac
tylu
s al
bila
bris
7
JAR
285
5 U
S PR
PR
H
atill
o 18
° 29
.175
' N
66°
49.9
12' W
Lept
odac
tylu
s al
bila
bris
7
JAR
285
6 U
S PR
PR
H
atill
o 18
° 29
.175
' N
66°
49.9
12' W
Lept
odac
tylu
s al
bila
bris
7
JAR
285
7 U
S PR
PR
H
atill
o 18
° 29
.175
' N
66°
49.9
12' W
Lept
odac
tylu
s al
bila
bris
7
JAR
285
8 U
S PR
PR
H
atill
o 18
° 29
.175
' N
66°
49.9
12' W
Lept
odac
tylu
s al
bila
bris
7
JAR
285
9 U
S PR
PR
H
atill
o 18
° 29
.175
' N
66°
49.9
12' W
Lept
odac
tylu
s al
bila
bris
7
JAR
286
0 U
S PR
PR
H
atill
o 18
° 29
.175
' N
66°
49.9
12' W
Lept
odac
tylu
s al
bila
bris
7
JAR
286
1 U
S PR
PR
H
atill
o 18
° 29
.175
' N
66°
49.9
12' W
Lept
odac
tylu
s al
bila
bris
7
JAR
286
2 U
S PR
PR
H
atill
o 18
° 29
.175
' N
66°
49.9
12' W
Lept
odac
tylu
s al
bila
bris
7
JAR
286
3 U
S PR
PR
H
atill
o 18
° 29
.175
' N
66°
49.9
12' W
Lept
odac
tylu
s al
bila
bris
8
JAR
287
8 U
S PR
PR
A
reci
bo
18°
22.7
30' N
66
° 44
.963
' W
Lept
odac
tylu
s al
bila
bris
8
JAR
287
9 U
S PR
PR
A
reci
bo
18°
22.7
30' N
66
° 44
.963
' W
Lept
odac
tylu
s al
bila
bris
8
JAR
288
0 U
S PR
PR
A
reci
bo
18°
22.7
30' N
66
° 44
.963
' W
35
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
8 JA
R 2
881
US
PR
PR
Are
cibo
18
° 22
.730
' N
66°
44.9
63' W
Lept
odac
tylu
s al
bila
bris
8
JAR
288
2 U
S PR
PR
A
reci
bo
18°
22.7
30' N
66
° 44
.963
' W
Lept
odac
tylu
s al
bila
bris
9
JAR
309
5 U
S PR
PR
Pe
ñuel
as
18°
04.5
86' N
66
° 43
.557
' W
Lept
odac
tylu
s al
bila
bris
9
JAR
309
6 U
S PR
PR
Pe
ñuel
as
18°
04.5
86' N
66
° 43
.557
' W
Lept
odac
tylu
s al
bila
bris
9
JAR
309
7 U
S PR
PR
Pe
ñuel
as
18°
04.5
86' N
66
° 43
.557
' W
Lept
odac
tylu
s al
bila
bris
9
JAR
309
8 U
S PR
PR
Pe
ñuel
as
18°
04.5
86' N
66
° 43
.557
' W
Lept
odac
tylu
s al
bila
bris
9
JAR
309
9 U
S PR
PR
Pe
ñuel
as
18°
04.5
86' N
66
° 43
.557
' W
Lept
odac
tylu
s al
bila
bris
9
JAR
310
0 U
S PR
PR
Pe
ñuel
as
18°
04.5
86' N
66
° 43
.557
' W
Lept
odac
tylu
s al
bila
bris
9
JAR
310
1 U
S PR
PR
Pe
ñuel
as
18°
04.5
86' N
66
° 43
.557
' W
Lept
odac
tylu
s al
bila
bris
9
JAR
310
2 U
S PR
PR
Pe
ñuel
as
18°
04.5
86' N
66
° 43
.557
' W
Lept
odac
tylu
s al
bila
bris
10
JA
R 3
037
US
PR
PR
Juan
a D
íaz
18°
09.0
87' N
66
° 32
.023
' W
Lept
odac
tylu
s al
bila
bris
10
JA
R 3
039
US
PR
PR
Juan
a D
íaz
18°
09.0
87' N
66
° 32
.023
' W
Lept
odac
tylu
s al
bila
bris
10
JA
R 3
040
US
PR
PR
Juan
a D
íaz
18°
09.0
87' N
66
° 32
.023
' W
Lept
odac
tylu
s al
bila
bris
10
JA
R 3
043
US
PR
PR
Juan
a D
íaz
18°
09.0
87' N
66
° 32
.023
' W
36
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
10
JAR
304
4 U
S PR
PR
Ju
ana
Día
z 18
° 09
.087
' N
66°
32.0
23' W
Lept
odac
tylu
s al
bila
bris
10
JA
R 3
045
US
PR
PR
Juan
a D
íaz
18°
09.0
87' N
66
° 32
.023
' W
Lept
odac
tylu
s al
bila
bris
10
JA
R 3
046
US
PR
PR
Juan
a D
íaz
18°
09.0
87' N
66
° 32
.023
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 2
993
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 2
994
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 2
995
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 2
996
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 2
997
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 2
998
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 2
999
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 3
000
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 3
001
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 3
002
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
Lept
odac
tylu
s al
bila
bris
11
JA
R 3
003
US
PR
PR
Cia
les
18°
14.8
05' N
66
° 31
.215
' W
37
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
11
JAR
300
4 U
S PR
PR
C
iale
s 18
° 14
.805
' N
66°
31.2
15' W
Lept
odac
tylu
s al
bila
bris
12
JA
R 2
709
US
PR
PR
Juan
a D
íaz
18°
00.3
95' N
66
° 29
.538
' W
Lept
odac
tylu
s al
bila
bris
12
JA
R 2
710
US
PR
PR
Juan
a D
íaz
18°
00.3
95' N
66
° 29
.538
' W
Lept
odac
tylu
s al
bila
bris
12
JA
R 2
711
US
PR
PR
Juan
a D
íaz
18°
00.3
95' N
66
° 29
.538
' W
Lept
odac
tylu
s al
bila
bris
12
JA
R 2
712
US
PR
PR
Juan
a D
íaz
18°
00.3
95' N
66
° 29
.538
' W
Lept
odac
tylu
s al
bila
bris
12
JA
R 2
713
US
PR
PR
Juan
a D
íaz
18°
00.3
95' N
66
° 29
.538
' W
Lept
odac
tylu
s al
bila
bris
12
JA
R 2
714
US
PR
PR
Juan
a D
íaz
18°
00.3
95' N
66
° 29
.538
' W
Lept
odac
tylu
s al
bila
bris
12
JA
R 2
715
US
PR
PR
Juan
a D
íaz
18°
00.3
95' N
66
° 29
.538
' W
Lept
odac
tylu
s al
bila
bris
12
JA
R 2
716
US
PR
PR
Juan
a D
íaz
18°
00.3
95' N
66
° 29
.538
' W
Lept
odac
tylu
s al
bila
bris
12
JA
R 2
717
US
PR
PR
Juan
a D
íaz
18°
00.3
95' N
66
° 29
.538
' W
Lept
odac
tylu
s al
bila
bris
12
JA
R 2
718
US
PR
PR
Juan
a D
íaz
18°
00.3
95' N
66
° 29
.538
' W
Lept
odac
tylu
s al
bila
bris
13
JA
R 2
699
US
PR
PR
Man
atí
18°
23.2
00' N
66
° 29
.119
' W
Lept
odac
tylu
s al
bila
bris
13
JA
R 2
700
US
PR
PR
Man
atí
18°
23.2
00' N
66
° 29
.119
' W
Lept
odac
tylu
s al
bila
bris
13
JA
R 2
701
US
PR
PR
Man
atí
18°
23.2
00' N
66
° 29
.119
' W
38
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
l abr
is
13
JAR
270
2 U
S PR
PR
M
anat
í 18
° 23
.200
' N
66°
29.1
19' W
Lept
odac
tylu
s al
bila
bris
13
JA
R 2
703
US
PR
PR
Man
atí
18°
23.2
00' N
66
° 29
.119
' W
Lept
odac
tylu
s al
bila
bris
13
JA
R 2
704
US
PR
PR
Man
atí
18°
23.2
00' N
66
° 29
.119
' W
Lept
odac
tylu
s al
bila
bris
13
JA
R 2
705
US
PR
PR
Man
atí
18°
23.2
00' N
66
° 29
.119
' W
Lept
odac
tylu
s al
bila
bris
13
JA
R 2
707
US
PR
PR
Man
atí
18°
23.2
00' N
66
° 29
.119
' W
Lept
odac
tylu
s al
bila
bris
13
JA
R 2
708
US
PR
PR
Man
atí
18°
23.2
00' N
66
° 29
.119
' W
Lept
odac
tylu
s al
bila
bris
14
JA
R 2
915
US
PR
PR
Dor
ado
18°
27.7
31' N
66
° 16
.399
' W
Lept
odac
tylu
s al
bila
bris
14
JA
R 2
916
US
PR
PR
Dor
ado
18°
27.7
31' N
66
° 16
.399
' W
Lept
odac
tylu
s al
bila
bris
14
JA
R 2
917
US
PR
PR
Dor
ado
18°
27.7
31' N
66
° 16
.399
' W
Lept
odac
tylu
s al
bila
bris
14
JA
R 2
918
US
PR
PR
Dor
ado
18°
27.7
31' N
66
° 16
.399
' W
Lept
odac
tylu
s al
bila
bris
14
JA
R 2
919
US
PR
PR
Dor
ado
18°
27.7
31' N
66
° 16
.399
' W
Lept
odac
tylu
s al
bila
bris
14
JA
R 2
920
US
PR
PR
Dor
ado
18°
27.7
31' N
66
° 16
.399
' W
Lept
odac
tylu
s al
bila
bris
14
JA
R 2
921
US
PR
PR
Dor
ado
18°
27.7
31' N
66
° 16
.399
' W
Lept
odac
tylu
s al
bila
bris
14
JA
R 2
922
US
PR
PR
Dor
ado
18°
27.7
31' N
66
° 16
.399
' W
39
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
14
JAR
292
3 U
S PR
PR
D
orad
o 18
° 27
.731
' N
66°
16.3
99' W
Lept
odac
tylu
s al
bila
bris
14
JA
R 2
924
US
PR
PR
Dor
ado
18°
27.7
31' N
66
° 16
.399
' W
Lept
odac
tylu
s al
bila
bris
15
JA
R 2
686
US
PR
PR
Aib
onito
18
° 06
.931
' N
66°
13.9
99' W
Lept
odac
tylu
s al
bila
bris
15
JA
R 2
687
US
PR
PR
Aib
onito
18
° 06
.931
' N
66°
13.9
99' W
Lept
odac
tylu
s al
bila
bris
15
JA
R 2
688
US
PR
PR
Aib
onito
18
° 06
.931
' N
66°
13.9
99' W
Lept
odac
tylu
s al
bila
bris
15
JA
R 2
689
US
PR
PR
Aib
onito
18
° 06
.931
' N
66°
13.9
99' W
Lept
odac
tylu
s al
bila
bris
15
JA
R 2
690
US
PR
PR
Aib
onito
18
° 06
.931
' N
66°
13.9
99' W
Lept
odac
tylu
s al
bila
bris
15
JA
R 2
691
US
PR
PR
Aib
onito
18
° 06
.931
' N
66°
13.9
99' W
Lept
odac
tylu
s al
bila
bris
15
JA
R 2
692
US
PR
PR
Aib
onito
18
° 06
.931
' N
66°
13.9
99' W
Lept
odac
tylu
s al
bila
bris
15
JA
R 2
693
US
PR
PR
Aib
onito
18
° 06
.931
' N
66°
13.9
99' W
Lept
odac
tylu
s al
bila
bris
15
JA
R 2
694
US
PR
PR
Aib
onito
18
° 06
.931
' N
66°
13.9
99' W
Lept
odac
tylu
s al
bila
bris
16
JA
R 2
685
US
PR
PR
Aib
onito
18
° 06
.574
' N
66°
13.5
72' W
Lept
odac
tylu
s al
bila
bris
17
JA
R 2
556
US
PR
PR
Toa
Alta
18
° 20
.222
' N
66°
12.8
48' W
Lept
odac
tylu
s al
bila
bris
17
JA
R 2
557
US
PR
PR
Toa
Alta
18
° 20
.222
' N
66°
12.8
48' W
40
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
17
JAR
255
8 U
S PR
PR
To
a A
lta
18°
20.2
22' N
66
° 12
.848
' W
Lept
odac
tylu
s al
bila
bris
17
JA
R 2
559
US
PR
PR
Toa
Alta
18
° 20
.222
' N
66°
12.8
48' W
Lept
odac
tylu
s al
bila
bris
17
JA
R 2
560
US
PR
PR
Toa
Alta
18
° 20
.222
' N
66°
12.8
48' W
Lept
odac
tylu
s al
bila
bris
17
JA
R 2
561
US
PR
PR
Toa
Alta
18
° 20
.222
' N
66°
12.8
48' W
Lept
odac
tylu
s al
bila
bris
17
JA
R 2
562
US
PR
PR
Toa
Alta
18
° 20
.222
' N
66°
12.8
48' W
Lept
odac
tylu
s al
bila
bris
17
JA
R 2
563
US
PR
PR
Toa
Alta
18
° 20
.222
' N
66°
12.8
48' W
Lept
odac
tylu
s al
bila
bris
17
JA
R 2
564
US
PR
PR
Toa
Alta
18
° 20
.222
' N
66°
12.8
48' W
Lept
odac
tylu
s al
bila
bris
17
JA
R 2
565
US
PR
PR
Toa
Alta
18
° 20
.222
' N
66°
12.8
48' W
Lept
odac
tylu
s al
bila
bris
18
JA
R 8
80
US
PR
PR
Gua
yam
a 17
° 57
.200
' N
66°
10.8
72' W
Lept
odac
tylu
s al
bila
bris
18
JA
R 8
81
US
PR
PR
Gua
yam
a 17
° 57
.200
' N
66°
10.8
72' W
Lept
odac
tylu
s al
bila
bris
18
JA
R 8
82
US
PR
PR
Gua
yam
a 17
° 57
.200
' N
66°
10.8
72' W
Lept
odac
tylu
s al
bila
bris
18
JA
R 8
83
US
PR
PR
Gua
yam
a 17
° 57
.200
' N
66°
10.8
72' W
Lept
odac
tylu
s al
bila
bris
18
JA
R 8
84
US
PR
PR
Gua
yam
a 17
° 57
.200
' N
66°
10.8
72' W
Lept
odac
tylu
s al
bila
bris
18
JA
R 8
85
US
PR
PR
Gua
yam
a 17
° 57
.200
' N
66°
10.8
72' W
41
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
18
JAR
886
U
S PR
PR
G
uaya
ma
17°
57.2
00' N
66
° 10
.872
' W
Lept
odac
tylu
s al
bila
bris
18
JA
R 8
87
US
PR
PR
Gua
yam
a 17
° 57
.200
' N
66°
10.8
72' W
Lept
odac
tylu
s al
bila
bris
19
JA
R 2
750
US
PR
PR
Gua
yam
a 17
° 57
.863
' N
66°
06.7
06' W
Lept
odac
tylu
s al
bila
bris
19
JA
R 2
751
US
PR
PR
Gua
yam
a 17
° 57
.863
' N
66°
06.7
06' W
Lept
odac
tylu
s al
bila
bris
19
JA
R 2
752
US
PR
PR
Gua
yam
a 17
° 57
.863
' N
66°
06.7
06' W
Lept
odac
tylu
s al
bila
bris
19
JA
R 2
753
US
PR
PR
Gua
yam
a 17
° 57
.863
' N
66°
06.7
06' W
Lept
odac
tylu
s al
bila
bris
19
JA
R 2
754
US
PR
PR
Gua
yam
a 17
° 57
.863
' N
66°
06.7
06' W
Lept
odac
tylu
s al
bila
bris
19
JA
R 2
755
US
PR
PR
Gua
yam
a 17
° 57
.863
' N
66°
06.7
06' W
Lept
odac
tylu
s al
bila
bris
19
JA
R 2
756
US
PR
PR
Gua
yam
a 17
° 57
.863
' N
66°
06.7
06' W
Lept
odac
tylu
s al
bila
bris
19
JA
R 2
757
US
PR
PR
Gua
yam
a 17
° 57
.863
' N
66°
06.7
06' W
Lept
odac
tylu
s al
bila
bris
19
JA
R 2
758
US
PR
PR
Gua
yam
a 17
° 57
.863
' N
66°
06.7
06' W
Lept
odac
tylu
s al
bila
bris
19
JA
R 2
759
US
PR
PR
Gua
yam
a 17
° 57
.863
' N
66°
06.7
06' W
Lept
odac
tylu
s al
bila
bris
20
JA
R 2
726
US
PR
PR
Agu
as B
uena
s 18
° 14
.403
' N
66°
06.2
39' W
Lept
odac
tylu
s al
bila
bris
20
JA
R 2
727
US
PR
PR
Agu
as B
uena
s 18
° 14
.403
' N
66°
06.2
39' W
42
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
20
JAR
272
8 U
S PR
PR
A
guas
Bue
nas
18°
14.4
03' N
66
° 06
.239
' W
Lept
odac
tylu
s al
bila
bris
20
JA
R 2
729
US
PR
PR
Agu
as B
uena
s 18
° 14
.403
' N
66°
06.2
39' W
Lept
odac
tylu
s al
bila
bris
20
JA
R 2
730
US
PR
PR
Agu
as B
uena
s 18
° 14
.403
' N
66°
06.2
39' W
Lept
odac
tylu
s al
bila
bris
20
JA
R 2
731
US
PR
PR
Agu
as B
uena
s 18
° 14
.403
' N
66°
06.2
39' W
Lept
odac
tylu
s al
bila
bris
20
JA
R 2
732
US
PR
PR
Agu
as B
uena
s 18
° 14
.403
' N
66°
06.2
39' W
Lept
odac
tylu
s al
bila
bris
20
JA
R 2
733
US
PR
PR
Agu
as B
uena
s 18
° 14
.403
' N
66°
06.2
39' W
Lept
odac
tylu
s al
bila
bris
20
JA
R 2
734
US
PR
PR
Agu
as B
uena
s 18
° 14
.403
' N
66°
06.2
39' W
Lept
odac
tylu
s al
bila
bris
20
JA
R 2
735
US
PR
PR
Agu
as B
uena
s 18
° 14
.403
' N
66°
06.2
39' W
Lept
odac
tylu
s al
bila
bris
21
JA
R 2
987
US
PR
PR
Cay
ey
18°
07.3
39' N
66
° 04
.698
' W
Lept
odac
tylu
s al
bila
bris
21
JA
R 2
988
US
PR
PR
Cay
ey
18°
07.3
39' N
66
° 04
.698
' W
Lept
odac
tylu
s al
bila
bris
21
JA
R 2
989
US
PR
PR
Cay
ey
18°
07.3
39' N
66
° 04
.698
' W
Lept
odac
tylu
s al
bila
bris
21
JA
R 2
990
US
PR
PR
Cay
ey
18°
07.3
39' N
66
° 04
.698
' W
Lept
odac
tylu
s al
bila
bris
21
JA
R 2
991
US
PR
PR
Cay
ey
18°
07.3
39' N
66
° 04
.698
' W
Lept
odac
tylu
s al
bila
bris
21
JA
R 2
992
US
PR
PR
Cay
ey
18°
07.3
39' N
66
° 04
.698
' W
43
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
22
JAR
298
2 U
S PR
PR
C
ayey
18
° 06
.578
' N
66°
04.2
89' W
Lept
odac
tylu
s al
bila
bris
22
JA
R 2
983
US
PR
PR
Cay
ey
18°
06.5
78' N
66
° 04
.289
' W
Lept
odac
tylu
s al
bila
bris
22
JA
R 2
984
US
PR
PR
Cay
ey
18°
06.5
78' N
66
° 04
.289
' W
Lept
odac
tylu
s al
bila
bris
22
JA
R 2
985
US
PR
PR
Cay
ey
18°
06.5
78' N
66
° 04
.289
' W
Lept
odac
tylu
s al
bila
bris
22
JA
R 2
986
US
PR
PR
Cay
ey
18°
06.5
78' N
66
° 04
.289
' W
Lept
odac
tylu
s al
bila
bris
23
JA
R 3
005
US
PR
PR
Gur
abo
18°
18.1
45' N
65
° 59
.238
' W
Lept
odac
tylu
s al
bila
bris
23
JA
R 3
006
US
PR
PR
Gur
abo
18°
18.1
45' N
65
° 59
.238
' W
Lept
odac
tylu
s al
bila
bris
23
JA
R 3
007
US
PR
PR
Gur
abo
18°
18.1
45' N
65
° 59
.238
' W
Lept
odac
tylu
s al
bila
bris
23
JA
R 3
008
US
PR
PR
Gur
abo
18°
18.1
45' N
65
° 59
.238
' W
Lept
odac
tylu
s al
bila
bris
23
JA
R 3
009
US
PR
PR
Gur
abo
18°
18.1
45' N
65
° 59
.238
' W
Lept
odac
tylu
s al
bila
bris
23
JA
R 3
010
US
PR
PR
Gur
abo
18°
18.1
45' N
65
° 59
.238
' W
Lept
odac
tylu
s al
bila
bris
23
JA
R 3
011
US
PR
PR
Gur
abo
18°
18.1
45' N
65
° 59
.238
' W
Lept
odac
tylu
s al
bila
bris
23
JA
R 3
012
US
PR
PR
Gur
abo
18°
18.1
45' N
65
° 59
.238
' W
Lept
odac
tylu
s al
bila
bris
23
JA
R 3
013
US
PR
PR
Gur
abo
18°
18.1
45' N
65
° 59
.238
' W
44
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
23
JAR
301
4 U
S PR
PR
G
urab
o 18
° 18
.145
' N
65°
59.2
38' W
Lept
odac
tylu
s al
bila
bris
24
JA
R 2
669
US
PR
PR
Car
olin
a 18
° 26
.549
' N
65°
57.4
15' W
Lept
odac
tylu
s al
bila
bris
24
JA
R 2
670
US
PR
PR
Car
olin
a 18
° 26
.549
' N
65°
57.4
15' W
Lept
odac
tylu
s al
bila
bris
24
JA
R 2
671
US
PR
PR
Car
olin
a 18
° 26
.549
' N
65°
57.4
15' W
Lept
odac
tylu
s al
bila
bris
24
JA
R 2
672
US
PR
PR
Car
olin
a 18
° 26
.549
' N
65°
57.4
15' W
Lept
odac
tylu
s al
bila
bris
24
JA
R 2
673
US
PR
PR
Car
olin
a 18
° 26
.549
' N
65°
57.4
15' W
Lept
odac
tylu
s al
bila
bris
24
JA
R 2
674
US
PR
PR
Car
olin
a 18
° 26
.549
' N
65°
57.4
15' W
Lept
odac
tylu
s al
bila
bris
24
JA
R 2
675
US
PR
PR
Car
olin
a 18
° 26
.549
' N
65°
57.4
15' W
Lept
odac
tylu
s al
bila
bris
24
JA
R 2
676
US
PR
PR
Car
olin
a 18
° 26
.549
' N
65°
57.4
15' W
Lept
odac
tylu
s al
bila
bris
24
JA
R 2
677
US
PR
PR
Car
olin
a 18
° 26
.549
' N
65°
57.4
15' W
Lept
odac
tylu
s al
bila
bris
24
JA
R 2
678
US
PR
PR
Car
olin
a 18
° 26
.549
' N
65°
57.4
15' W
Lept
odac
tylu
s al
bila
bris
25
JA
R 2
38
US
PR
PR
Mau
nabo
18
° 01
.623
' N
65°
54.8
88' W
Lept
odac
tylu
s al
bila
bris
25
JA
R 2
39
US
PR
PR
Mau
nabo
18
° 01
.623
' N
65°
54.8
88' W
Lept
odac
tylu
s al
bila
bris
25
JA
R 2
40
US
PR
PR
Mau
nabo
18
° 01
.623
' N
65°
54.8
88' W
45
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
25
JAR
241
U
S PR
PR
M
auna
bo
18°
01.6
23' N
65
° 54
.888
' W
Lept
odac
tylu
s al
bila
bris
26
JA
R 2
577
US
PR
PR
Mau
nabo
18
° 01
.586
' N
65°
54.8
22' W
Lept
odac
tylu
s al
bila
bris
26
JA
R 2
578
US
PR
PR
Mau
nabo
18
° 01
.586
' N
65°
54.8
22' W
Lept
odac
tylu
s al
bila
bris
26
JA
R 2
579
US
PR
PR
Mau
nabo
18
° 01
.586
' N
65°
54.8
22' W
Lept
odac
tylu
s al
bila
bris
26
JA
R 2
580
US
PR
PR
Mau
nabo
18
° 01
.586
' N
65°
54.8
22' W
Lept
odac
tylu
s al
bila
bris
26
JA
R 2
581
US
PR
PR
Mau
nabo
18
° 01
.586
' N
65°
54.8
22' W
Lept
odac
tylu
s al
bila
bris
26
JA
R 2
582
US
PR
PR
Mau
nabo
18
° 01
.586
' N
65°
54.8
22' W
Lept
odac
tylu
s al
bila
bris
27
JA
R 2
960
US
PR
PR
Junc
os
18°
14.0
06' N
65
° 52
.088
' W
Lept
odac
tylu
s al
bila
bris
27
JA
R 2
961
US
PR
PR
Junc
os
18°
14.0
06' N
65
° 52
.088
' W
Lept
odac
tylu
s al
bila
bris
27
JA
R 2
962
US
PR
PR
Junc
os
18°
14.0
06' N
65
° 52
.088
' W
Lept
odac
tylu
s al
bila
bris
27
JA
R 2
963
US
PR
PR
Junc
os
18°
14.0
06' N
65
° 52
.088
' W
Lept
odac
tylu
s al
bila
bris
27
JA
R 2
964
US
PR
PR
Junc
os
18°
14.0
06' N
65
° 52
.088
' W
Lept
odac
tylu
s al
bila
bris
27
JA
R 2
965
US
PR
PR
Junc
os
18°
14.0
06' N
65
° 52
.088
' W
Lept
odac
tylu
s al
bila
bris
27
JA
R 2
966
US
PR
PR
Junc
os
18°
14.0
06' N
65
° 52
.088
' W
46
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
27
JAR
296
7 U
S PR
PR
Ju
ncos
18
° 14
.006
' N
65°
52.0
88' W
Lept
odac
tylu
s al
bila
bris
27
JA
R 2
968
US
PR
PR
Junc
os
18°
14.0
06' N
65
° 52
.088
' W
Lept
odac
tylu
s al
bila
bris
27
JA
R 2
969
US
PR
PR
Junc
os
18°
14.0
06' N
65
° 52
.088
' W
Lept
odac
tylu
s al
bila
bris
28
JA
R 2
929
US
PR
PR
Río
Gra
nde
18°
23.4
27' N
65
° 46
.837
' W
Lept
odac
tylu
s al
bila
bris
28
JA
R 2
930
US
PR
PR
Río
Gra
nde
18°
23.4
27' N
65
° 46
.837
' W
Lept
odac
tylu
s al
bila
bris
28
JA
R 2
931
US
PR
PR
Río
Gra
nde
18°
23.4
27' N
65
° 46
.837
' W
Lept
odac
tylu
s al
bila
bris
28
JA
R 2
932
US
PR
PR
Río
Gra
nde
18°
23.4
27' N
65
° 46
.837
' W
Lept
odac
tylu
s al
bila
bris
28
JA
R 2
933
US
PR
PR
Río
Gra
nde
18°
23.4
27' N
65
° 46
.837
' W
Lept
odac
tylu
s al
bila
bris
28
JA
R 2
934
US
PR
PR
Río
Gra
nde
18°
23.4
27' N
65
° 46
.837
' W
Lept
odac
tylu
s al
bila
bris
28
JA
R 2
935
US
PR
PR
Río
Gra
nde
18°
23.4
27' N
65
° 46
.837
' W
Lept
odac
tylu
s al
bila
bris
28
JA
R 2
936
US
PR
PR
Río
Gra
nde
18°
23.4
27' N
65
° 46
.837
' W
Lept
odac
tylu
s al
bila
bris
28
JA
R 2
937
US
PR
PR
Río
Gra
nde
18°
23.4
27' N
65
° 46
.837
' W
Lept
odac
tylu
s al
bila
bris
28
JA
R 2
938
US
PR
PR
Río
Gra
nde
18°
23.4
27' N
65
° 46
.837
' W
Lept
odac
tylu
s al
bila
bris
29
JA
R 2
636
US
PR
PR
Faja
rdo
18°
16.2
46' N
65
° 42
.882
' W
47
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
a lbi
labr
is
29
JAR
263
7 U
S PR
PR
Fa
jard
o 18
° 16
.246
' N
65°
42.8
82' W
Lept
odac
tylu
s al
bila
bris
29
JA
R 2
638
US
PR
PR
Faja
rdo
18°
16.2
46' N
65
° 42
.882
' W
Lept
odac
tylu
s al
bila
bris
29
JA
R 2
639
US
PR
PR
Faja
rdo
18°
16.2
46' N
65
° 42
.882
' W
Lept
odac
tylu
s al
bila
bris
29
JA
R 2
640
US
PR
PR
Faja
rdo
18°
16.2
46' N
65
° 42
.882
' W
Lept
odac
tylu
s al
bila
bris
29
JA
R 2
641
US
PR
PR
Faja
rdo
18°
16.2
46' N
65
° 42
.882
' W
Lept
odac
tylu
s al
bila
bris
29
JA
R 2
642
US
PR
PR
Faja
rdo
18°
16.2
46' N
65
° 42
.882
' W
Lept
odac
tylu
s al
bila
bris
29
JA
R 2
643
US
PR
PR
Faja
rdo
18°
16.2
46' N
65
° 42
.882
' W
Lept
odac
tylu
s al
bila
bris
29
JA
R 2
644
US
PR
PR
Faja
rdo
18°
16.2
46' N
65
° 42
.882
' W
Lept
odac
tylu
s al
bila
bris
29
JA
R 2
645
US
PR
PR
Faja
rdo
18°
16.2
46' N
65
° 42
.882
' W
Lept
odac
tylu
s al
bila
bris
30
JA
R 3
049
US
PR
VI
VI
18°
06.0
88' N
65
° 32
.347
' W
Lept
odac
tylu
s al
bila
bris
30
JA
R 3
050
US
PR
VI
VI
18°
06.0
88' N
65
° 32
.347
' W
Lept
odac
tylu
s al
bila
bris
30
JA
R 3
051
US
PR
VI
VI
18°
06.0
88' N
65
° 32
.347
' W
Lept
odac
tylu
s al
bila
bris
30
JA
R 3
052
US
PR
VI
VI
18°
06.0
88' N
65
° 32
.347
' W
Lept
odac
tylu
s al
bila
bris
30
JA
R 3
053
US
PR
VI
VI
18°
06.0
88' N
65
° 32
.347
' W
48
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
30
JAR
305
4 U
S PR
V
I V
I 18
° 06
.088
' N
65°
32.3
47' W
Lept
odac
tylu
s al
bila
bris
30
JA
R 3
055
US
PR
VI
VI
18°
06.0
88' N
65
° 32
.347
' W
Lept
odac
tylu
s al
bila
bris
30
JA
R 3
056
US
PR
VI
VI
18°
06.0
88' N
65
° 32
.347
' W
Lept
odac
tylu
s al
bila
bris
30
JA
R 3
057
US
PR
VI
VI
18°
06.0
88' N
65
° 32
.347
' W
Lept
odac
tylu
s al
bila
bris
30
JA
R 3
058
US
PR
VI
VI
18°
06.0
88' N
65
° 32
.347
' W
Lept
odac
tylu
s al
bila
bris
31
JA
R 3
059
US
PR
VI
VI
18°
07.2
61' N
65
° 26
.204
' W
Lept
odac
tylu
s al
bila
bris
32
JA
R 2
943
US
PR
CU
C
U
18°
18.6
31' N
65
° 17
.969
' W
Lept
odac
tylu
s al
bila
bris
32
JA
R 2
944
US
PR
CU
C
U
18°
18.6
31' N
65
° 17
.969
' W
Lept
odac
tylu
s al
bila
bris
32
JA
R 2
945
US
PR
CU
C
U
18°
18.6
31' N
65
° 17
.969
' W
Lept
odac
tylu
s al
bila
bris
32
JA
R 2
946
US
PR
CU
C
U
18°
18.6
31' N
65
° 17
.969
' W
Lept
odac
tylu
s al
bila
bris
32
JA
R 2
947
US
PR
CU
C
U
18°
18.6
31' N
65
° 17
.969
' W
Lept
odac
tylu
s al
bila
bris
32
JA
R 2
948
US
PR
CU
C
U
18°
18.6
31' N
65
° 17
.969
' W
Lept
odac
tylu
s al
bila
bris
32
JA
R 2
949
US
PR
CU
C
U
18°
18.6
31' N
65
° 17
.969
' W
Lept
odac
tylu
s al
bila
bris
32
JA
R 2
950
US
PR
CU
C
U
18°
18.6
31' N
65
° 17
.969
' W
49
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
32
JAR
295
1 U
S PR
C
U
CU
18
° 18
.631
' N
65°
17.9
69' W
Lept
odac
tylu
s al
bila
bris
32
JA
R 2
952
US
PR
CU
C
U
18°
18.6
31' N
65
° 17
.969
' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
131
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
132
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
133
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
134
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
135
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
136
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
137
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
138
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
139
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
140
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
33
JA
R 3
141
US
USV
I ST
T —
18
° 20
.401
' N
64°
58.1
24' W
Lept
odac
tylu
s al
bila
bris
34
B
B 7
788
US
USV
I ST
T —
18
° 19
.375
' N
64°
54.6
49' W
50
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
35
JAR
318
1 U
S U
SVI
STC
—
17
° 43
.898
' N
64°
51.6
45' W
Lept
odac
tylu
s al
bila
bris
35
JA
R 3
182
US
USV
I ST
C
—
17°
43.8
98' N
64
° 51
.645
' W
Lept
odac
tylu
s al
bila
bris
35
JA
R 3
183
US
USV
I ST
C
—
17°
43.8
98' N
64
° 51
.645
' W
Lept
odac
tylu
s al
bila
bris
35
JA
R 3
184
US
USV
I ST
C
—
17°
43.8
98' N
64
° 51
.645
' W
Lept
odac
tylu
s al
bila
bris
35
JA
R 3
185
US
USV
I ST
C
—
17°
43.8
98' N
64
° 51
.645
' W
Lept
odac
tylu
s al
bila
bris
36
JA
R 3
170
US
USV
I ST
C
—
17°
45.5
44' N
64
° 45
.912
' W
Lept
odac
tylu
s al
bila
bris
36
JA
R 3
171
US
USV
I ST
C
—
17°
45.5
44' N
64
° 45
.912
' W
Lept
odac
tylu
s al
bila
bris
36
JA
R 3
172
US
USV
I ST
C
—
17°
45.5
44' N
64
° 45
.912
' W
Lept
odac
tylu
s al
bila
bris
36
JA
R 3
173
US
USV
I ST
C
—
17°
45.5
44' N
64
° 45
.912
' W
Lept
odac
tylu
s al
bila
bris
36
JA
R 3
174
US
USV
I ST
C
—
17°
45.5
44' N
64
° 45
.912
' W
Lept
odac
tylu
s al
bila
bris
36
JA
R 3
175
US
USV
I ST
C
—
17°
45.5
44' N
64
° 45
.912
' W
Lept
odac
tylu
s al
bila
bris
36
JA
R 3
176
US
USV
I ST
C
—
17°
45.5
44' N
64
° 45
.912
' W
Lept
odac
tylu
s al
bila
bris
36
JA
R 3
177
US
USV
I ST
C
—
17°
45.5
44' N
64
° 45
.912
' W
Lept
odac
tylu
s al
bila
bris
36
JA
R 3
178
US
USV
I ST
C
—
17°
45.5
44' N
64
° 45
.912
' W
51
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
36
JAR
317
9 U
S U
SVI
STC
—
17
° 45
.544
' N
64°
45.9
12' W
Lept
odac
tylu
s al
bila
bris
36
JA
R 3
180
US
USV
I ST
C
—
17°
45.5
44' N
64
° 45
.912
' W
Lept
odac
tylu
s al
bila
bris
37
B
B 7
847
US
USV
I ST
C
—
17°
44.2
93' N
64
° 42
.910
' W
Lept
odac
tylu
s al
bila
bris
37
B
B 7
848
US
USV
I ST
C
—
17°
44.2
93' N
64
° 42
.910
' W
Lept
odac
tylu
s al
bila
bris
38
JA
R 3
247
US
USV
I ST
J —
18
° 19
.668
' N
64°
47.4
81' W
Lept
odac
tylu
s al
bila
bris
38
JA
R 3
248
US
USV
I ST
J —
18
° 19
.668
' N
64°
47.4
81' W
Lept
odac
tylu
s al
bila
bris
38
JA
R 3
249
US
USV
I ST
J —
18
° 19
.668
' N
64°
47.4
81' W
Lept
odac
tylu
s al
bila
bris
38
JA
R 3
250
US
USV
I ST
J —
18
° 19
.668
' N
64°
47.4
81' W
Lept
odac
tylu
s al
bila
bris
38
JA
R 3
251
US
USV
I ST
J —
18
° 19
.668
' N
64°
47.4
81' W
Lept
odac
tylu
s al
bila
bris
38
JA
R 3
252
US
USV
I ST
J —
18
° 19
.668
' N
64°
47.4
81' W
Lept
odac
tylu
s al
bila
bris
38
JA
R 3
253
US
USV
I ST
J —
18
° 19
.668
' N
64°
47.4
81' W
Lept
odac
tylu
s al
bila
bris
38
JA
R 3
254
US
USV
I ST
J —
18
° 19
.668
' N
64°
47.4
81' W
Lept
odac
tylu
s al
bila
bris
38
JA
R 3
255
US
USV
I ST
J —
18
° 19
.668
' N
64°
47.4
81' W
Lept
odac
tylu
s al
bila
bris
38
JA
R 3
256
US
USV
I ST
J —
18
° 19
.668
' N
64°
47.4
81' W
52
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
39
JAR
323
9 U
K
BV
I JV
D
—
18°
26.7
86' N
64
° 45
.078
' W
Lept
odac
tylu
s al
bila
bris
39
JA
R 3
240
UK
B
VI
JVD
—
18
° 26
.786
' N
64°
45.0
78' W
Lept
odac
tylu
s al
bila
bris
39
JA
R 3
241
UK
B
VI
JVD
—
18
° 26
.786
' N
64°
45.0
78' W
Lept
odac
tylu
s al
bila
bris
39
JA
R 3
242
UK
B
VI
JVD
—
18
° 26
.786
' N
64°
45.0
78' W
Lept
odac
tylu
s al
bila
bris
39
JA
R 3
243
UK
B
VI
JVD
—
18
° 26
.786
' N
64°
45.0
78' W
Lept
odac
tylu
s al
bila
bris
39
JA
R 3
244
UK
B
VI
JVD
—
18
° 26
.786
' N
64°
45.0
78' W
Lept
odac
tylu
s al
bila
bris
39
JA
R 3
245
UK
B
VI
JVD
—
18
° 26
.786
' N
64°
45.0
78' W
Lept
odac
tylu
s al
bila
bris
39
JA
R 3
246
UK
B
VI
JVD
—
18
° 26
.786
' N
64°
45.0
78' W
Lept
odac
tylu
s al
bila
bris
40
B
B 7
521
UK
B
VI
JVD
—
18
° 26
.861
' N
64°
44.4
04' W
Lept
odac
tylu
s al
bila
bris
40
B
B 7
522
UK
B
VI
JVD
—
18
° 26
.861
' N
64°
44.4
04' W
Lept
odac
tylu
s al
bila
bris
40
B
B 7
523
UK
B
VI
JVD
—
18
° 26
.861
' N
64°
44.4
04' W
Lept
odac
tylu
s al
bila
bris
41
B
B 7
491
UK
B
VI
TO
—
18°
25.3
51' N
64
° 38
.671
' W
Lept
odac
tylu
s al
bila
bris
42
JA
R 3
233
UK
B
VI
TO
—
18°
25.9
19' N
64
° 38
.268
' W
Lept
odac
tylu
s al
bila
bris
42
JA
R 3
234
UK
B
VI
TO
—
18°
25.9
19' N
64
° 38
.268
' W
53
Tab
le 1
. Con
tinue
d.
Spec
ies
Popu
latio
n nu
mbe
r Fi
eld
nu
mbe
r C
ount
ry
Stat
e Is
land
M
unic
ipal
ity
Lat
itude
(h
) L
atitu
de
(min
) L
ongi
tude
(h
) L
ongi
tude
(m
in)
Le
ptod
acty
lus
albi
labr
is
42
JAR
323
5 U
K
BV
I TO
—
18
° 25
.919
' N
64°
38.2
68' W
Lept
odac
tylu
s al
bila
bris
42
JA
R 3
236
UK
B
VI
TO
—
18°
25.9
19' N
64
° 38
.268
' W
Lept
odac
tylu
s al
bila
bris
42
JA
R 3
237
UK
B
VI
TO
—
18°
25.9
19' N
64
° 38
.268
' W
Lept
odac
tylu
s al
bila
bris
42
JA
R 3
238
UK
B
VI
TO
—
18°
25.9
19' N
64
° 38
.268
' W
54
Table 2. Phylogroup and associated population(s), population number(s), and haplotypes of
Leptodactylus albilabris. Phylogroups (PG) are clades supported by ≥ 90% posterior probability
values in the Bayesian Inference tree estimated for 140 unique cytochrome b haplotypes of L.
albilabris (Figure 2).
Phylogroup Population(s) Population number(s) Haplotypes
PG 1 Culebra 32 43, 60
PG 2 Carolina, Fajardo 24, 29 116, 126, 130
PG 3 Juana Díaz 2, Guayama 1 12, 18 68, 81
PG 4 Hatillo, Juana Díaz 1, Ciales 7, 10, 11 63, 82
PG 5 Aibonito 1, Aguas Buenas, Cayey 1 15, 20, 21 61, 100
PG 6 Ciales 11 21, 48
PG 7 Moca, Isabela, Yauco 2, 3, 6 17, 26, 73, 74
PG 8 Mayagüez, Guánica, Yauco, Hatillo, Peñuelas,
Juana Díaz, Ciales, Juana Díaz, Manatí,
Aibonito 2, Toa Alta, Guayama 1, Guayama 2,
Aguas Buenas, Cayey 1, Cayey 2, Gurabo,
Maunabo 2, Juncos, Culebra
4, 5, 6, 7, 9, 10,
11, 12, 13, 16,
17, 18, 19, 20,
21, 22, 23, 26,
27, 32
85, 91, 103,
104, 114, 115,
118, 127, 128,
129, 131, 132,
133, 136, 137,
138, 139, 140
PG 9 Cabo Rojo, Hatillo 1, 7 110, 119
PG 10 Manatí, Aguas Buenas 13, 20 83, 93
55
Table 2. Continued.
Phylogroup Population(s) Population number(s) Haplotypes
PG 11 Aibonito 1, Guayama 1, Guayama 2,
Aguas Buenas, Cayey 1, Cayey 2, Gurabo,
Carolina, Maunabo 1, Maunabo 2, Juncos,
Fajardo, Vieques 1, Vieques 2
15, 18, 19, 20, 21,
22, 23, 24, 25, 26,
27, 29, 30, 31
1, 2, 3, 4, 72,
84, 88, 90, 95,
97, 98, 99, 101,
102, 105, 134
PG 12 Isabela, Arecibo 3, 8 40, 42
PG 13 Dorado 14 12, 34
PG 14 Juana Díaz, Ciales, Manatí, Toa Alta,
Cayey 1
10, 11, 13, 17, 21 20, 38, 54
PG 15 Moca 2 19, 23
PG 16 Cabo Rojo, Guánica, Juncos, Río Grande 1, 5, 27, 28 9, 29, 31, 32, 76
PG 17 Guayama 2, Gurabo, Juncos, Río Grande 19, 23, 27, 28 14, 15, 36, 49,
69, 75
PG 18 Carolina, Río Grande 24, 28 55, 57
56
Table 3. Island, population number, municipality, and unique cytochrome b haplotype(s)
detected in 42 populations of Leptodactylus albilabris.
Island Population number Municipality Haplotype
Puerto Rico 1 Cabo Rojo 9, 11, 29, 31, 32, 37, 44, 110
Puerto Rico 2 Moca 19, 23, 24, 35, 73, 92, 96
Puerto Rico 3 Isabela 7, 17, 25, 41, 42, 74
Puerto Rico 4 Mayagüez 18, 70, 96, 127
Puerto Rico 5 Guánica 9, 11, 16, 29, 44, 46, 115
Puerto Rico 6 Yauco 26, 47, 71, 96, 115, 117, 120
Puerto Rico 7 Hatillo 82, 86, 87, 96, 115, 119, 125
Puerto Rico 8 Arecibo 35, 40, 94, 96
Puerto Rico 9 Peñuelas 115, 138
Puerto Rico 10 Juana Díaz 1 20, 38, 63, 115
Puerto Rico 11 Ciales 21, 38, 39, 48, 63, 96, 103, 106, 139
Puerto Rico 12 Juana Díaz 2 11, 68, 85, 96, 115, 128, 140
Puerto Rico 13 Manatí 20, 52, 71, 83, 86, 96, 133
Puerto Rico 14 Dorado 12, 27, 34, 51, 52, 123
Puerto Rico 15 Aibonito 1 1, 11, 96, 100, 124
Puerto Rico 16 Aibonito 2 129
Puerto Rico 17 Toa Alta 13, 20, 27, 52, 91, 115, 124, 133
Puerto Rico 18 Guayama 1 1, 81, 115, 131
57
Table 3. Continued.
Island Population number Municipality Haplotype
Puerto Rico 19 Guayama 2 1, 11, 15, 22, 75, 115
Puerto Rico 20 Aguas Buenas 53, 61, 66, 72, 84, 88, 89, 91, 93, 134
Puerto Rico 21 Cayey 1 2, 6, 33, 54, 91, 100, 137
Puerto Rico 22 Cayey 2 1, 79, 118, 135
Puerto Rico 23 Gurabo 1, 49, 78, 80, 96, 115
Puerto Rico 24 Carolina 6, 50, 55, 67, 95, 105, 126
Puerto Rico 25 Maunabo 1 6, 102, 112, 122
Puerto Rico 26 Maunabo 2 1, 101, 114, 115, 132
Puerto Rico 27 Juncos 1, 6, 11, 25, 65, 69, 76, 77, 113, 115
Puerto Rico 28 Río Grande 6, 9, 14, 36, 56, 57, 58, 121
Puerto Rico 29 Fajardo 1, 11, 59, 62, 90, 116, 126, 130
Vieques 1 30 – 3, 6, 64, 97, 98, 99
Vieques 2 31 – 4
Culebra 32 – 43, 45, 60, 104, 136
Saint Thomas 1 33 – 10, 28
Saint Thomas 2 34 – 8
Saint Croix 1 35 – 10
Saint Croix 2 36 – 10, 30
Saint Croix 3 37 – 10
58
Table 3. Continued.
Island Population number Municipality Haplotype
Saint John 38 — 5, 6, 109
Jost Van Dyke 39 — 6, 111
Jost Van Dyke 40 — 6
Tortola 41 — 6
Tortola 42 — 6, 107, 108
59
Tab
le 4
. Isl
and,
pop
ulat
ion
num
ber,
mun
icip
ality
, sam
ple
size
, num
ber o
f uni
que
hapl
otyp
es, h
aplo
type
div
ersi
ty (H
d), a
nd n
ucle
otid
e
dive
rsity
(πd)
est
imat
ed u
sing
a c
ytoc
hrom
e b
frag
men
t (82
8 ba
se p
airs
) fro
m 3
22 sp
ecim
ens o
f Lep
toda
ctyl
us a
lbila
bris
from
42
loca
litie
s acr
oss t
he P
uerto
Ric
an B
ank.
I es
timat
ed H
d and
πd u
sing
AR
ELEQ
UIN
3.5
.2.2
(Exc
offie
r and
Lis
cher
201
0) fo
r loc
aliti
es
repr
esen
ted
by ≥
5 in
divi
dual
s.
Isla
nd
Popu
latio
n nu
mbe
r M
unic
ipal
ity
Sam
ple
size
Num
ber
of
uniq
ue
hapl
otyp
es
Hap
loty
pe
dive
rsity
(Hd)
N
ucle
otid
e di
vers
ity (π
)
Puer
to R
ico
1 C
abo
Roj
o 10
8
0.96
+/-
0.06
0.
0039
+/-
0.00
25
Puer
to R
ico
2 M
oca
8 7
0.96
+/-
0.08
0.
0051
+/-
0.00
32
Puer
to R
ico
3 Is
abel
a 10
6
0.89
+/-
0.08
0.
0059
+/-
0.00
35
Puer
to R
ico
4 M
ayag
üez
5 4
0.90
+/-
0.16
0.
0053
+/-
0.00
37
Puer
to R
ico
5 G
uáni
ca
10
7 0.
91 +
/- 0.
08
0.00
47 +
/- 0.
0029
Puer
to R
ico
6 Y
auco
10
7
0.91
+/-
0.09
0.
0048
+/-
0.00
29
Puer
to R
ico
7 H
atill
o 10
7
0.91
+/-
0.10
0.
0063
+/-
0.00
38
Puer
to R
ico
8 A
reci
bo
5 4
0.90
+/-
0.16
0.
0051
+/-
0.00
35
Puer
to R
ico
9 Pe
ñuel
as
8 2
0.25
+/-
0.18
0.
0006
+/-
0.00
06
60
Tab
le 4
. Con
tinue
d.
Isla
nd
Popu
latio
n nu
mbe
r M
unic
ipal
ity
Sam
ple
size
Num
ber
of
uniq
ue
hapl
otyp
es
Hap
loty
pe
dive
rsity
(Hd)
N
ucle
otid
e di
vers
ity (π
)
Puer
to R
ico
10
Juan
a D
íaz
1
7 4
0.86
+/-
0.10
0.
0078
+/-
0.00
48
Puer
to R
ico
11
Cia
les
12
9 0.
95 +
/- 0
.05
0.00
88 +
/- 0.
0050
Puer
to R
ico
12
Juan
a D
íaz
2 10
7
0.87
+/-
0.11
0.
0045
+/-
0.00
28
Puer
to R
ico
13
Man
atí
9 7
0.92
+/-
0.09
0.
0074
+/-
0.00
44
Puer
to R
ico
14
Dor
ado
10
6 0.
89 +
/- 0.
08
0.00
85 +
/- 0.
0049
Puer
to R
ico
15
Aib
onito
1
9 5
0.86
+/-
0.09
0.
0058
+/-
0.00
35
Puer
to R
ico
16
Aib
onito
2
1 —
—
—
Puer
to R
ico
17
Toa
Alta
10
8
0.96
+/-
0.06
0.
0073
+/-
0.00
43
Puer
to R
ico
18
Gua
yam
a 1
8 4
0.82
+/-
0.10
0.
0058
+/-
0.00
36
Puer
to R
ico
19
Gua
yam
a 2
10
6 0.
89 +
/- 0.
08
0.00
53 +
/- 0.
0032
Puer
to R
ico
20
Agu
as B
uena
s 10
10
1.
00 +
/- 0.
04
0.01
21 +
/- 0.
0068
Puer
to R
ico
21
Cay
ey 1
6
6 1.
00 +
/- 0.
10
0.01
10 +
/- 0.
0068
Puer
to R
ico
22
Cay
ey 2
5
5 1.
00 +
/- 0.
13
0.01
04 +
/- 0.
0068
61
Tab
le 4
. Con
tinue
d.
Isla
nd
Popu
latio
n nu
mbe
r M
unic
ipal
ity
Sam
ple
size
Num
ber
of
uniq
ue
hapl
otyp
es
Hap
loty
pe
dive
rsity
(Hd)
N
ucle
otid
e di
vers
ity (π
)
Puer
to R
ico
23
Gur
abo
10
6 0.
87 +
/- 0.
09
0.00
92 +
/- 0.
0053
Puer
to R
ico
24
Car
olin
a 10
7
0.93
+/-
0.06
0.
0111
+/-
0.00
63
Puer
to R
ico
25
Mau
nabo
1
4 4
—
—
Puer
to R
ico
26
Mau
nabo
2
6 5
0.93
+/-
0.12
0.
0066
+/-
0.00
43
Puer
to R
ico
27
Junc
os
10
10
1.00
+/-
0.04
0.
0072
+/-
0.00
42
Puer
to R
ico
28
Río
Gra
nde
10
8 0.
96 +
/- 0.
06
0.00
82 +
/- 0.
0048
Puer
to R
ico
29
Faja
rdo
10
8 0.
93 +
/- 0.
08
0.01
04 +
/- 0.
0060
Vie
ques
1
30
—
10
6 0.
89 +
/- 0.
08
0.00
74 +
/- 0.
0044
Vie
ques
2
31
—
1 —
—
—
Cul
ebra
32
—
10
5
0.84
+/-
0.0
8 0.
0097
+/-
0.00
56
Sain
t Tho
mas
1
33
—
11
2 0.
51 +
/- 0
.10
0.00
00 +
/- 0.
0000
Sain
t Tho
mas
2
34
—
1 —
—
—
62
Tab
le 4
. Con
tinue
d.
Isla
nd
Popu
latio
n nu
mbe
r M
unic
ipal
ity
Sam
ple
size
Num
ber
of
uniq
ue
hapl
otyp
es
Hap
loty
pe
dive
rsity
(Hd)
N
ucle
otid
e di
vers
ity (π
)
Sain
t Cro
ix 1
35
—
5
1 0.
00 +
/- 0.
00
0.00
00 +
/- 0.
0000
Sain
t Cro
ix 2
36
—
11
2
0.18
+/-
0.1
4 0.
0002
+/-
0.00
03
Sain
t Cro
ix 3
37
—
2
—
—
—
Sain
t Joh
n 38
—
10
3
0.69
+/-
0.10
0.
0011
+/-
0.00
09
Jost
Van
Dyk
e 1
39
—
8 2
0.25
+/-
0.18
0.
0003
+/-
0.00
04
Jost
Van
Dyk
e 2
40
—
2 —
—
—
Torto
la 1
41
—
1
—
—
—
Torto
la 2
42
—
6
3 0.
60 +
/- 0.
22
0.00
04 +
/- 0.
0005
63
Tab
le 5
. Pai
rwis
e F s
t val
ues a
mon
g 35
pop
ulat
ions
of L
epto
dact
ylus
alb
ilabr
is re
pres
ente
d by
≥ 5
indi
vidu
als.
Val
ues w
ere
estim
ated
from
322
cyt
ochr
ome
b se
quen
ces.
Popu
latio
ns a
re li
sted
from
wes
t to
east
.
Popu
latio
n C
abo
Roj
o M
oca
Isab
ela
May
agüe
z G
uáni
ca
Yau
co
Hat
illo
Are
cibo
C
abo
Roj
o 0.
00
—
—
—
—
—
—
—
Moc
a 0.
0402
0.
0 —
—
—
—
—
—
Isab
ela
0.07
78
0.07
47
0.0
—
—
—
—
—
May
agüe
z 0.
0687
0.
0159
0.
1063
0.
0 —
—
—
—
Guá
nica
0.
0071
0.
0632
0.
1000
0.
0937
0.
0 —
—
—
Yau
co
0.06
67
0.03
91
0.10
00
0.01
50
0.07
03
0.0
—
—
Hat
illo
0.06
67
0.05
13
0.10
00
0.05
60
0.03
07
0.04
09
0.0
—
Are
cibo
0.
0687
0.
0159
0.
1063
0.
0217
0.
0937
0.
0560
0.
0752
0.
0
Peñu
elas
0.
3748
0.
3929
0.
4092
0.
4815
0.
2748
0.
3420
0.
1924
0.
4815
Juan
a D
íaz
1 0.
0907
0.
0880
0.
1260
0.
1231
0.
0607
0.
0882
0.
0315
0.
1231
Cia
les
0.04
50
0.02
04
0.07
75
0.00
28
0.06
67
0.03
46
0.05
09
0.03
69
Juan
a D
íaz
2 0.
0797
0.
0746
0.
1222
0.
0821
0.
0232
0.
0544
-0
.021
7 0.
1010
64
Tab
le 5
. Con
tinue
d.
Popu
latio
n C
abo
Roj
o M
oca
Isab
ela
May
agüe
z G
uáni
ca
Yau
co
Hat
illo
Are
cibo
M
anat
í 0.
0636
0.
0467
0.
0975
0.
0485
0.
0862
0.
0324
0.
0654
0.
0701
Dor
ado
0.07
78
0.07
47
0.11
11
0.10
63
0.10
00
0.10
00
0.10
00
0.10
63
Aib
onito
0.
0703
0.
0622
0.
1248
0.
0358
0.
0933
0.
0723
0.
0933
0.
0808
Toa
Alta
0.
0444
0.
0402
0.
0778
0.
0687
0.
0278
0.
0476
0.
0071
0.
0687
Gua
yam
a 1
0.10
90
0.10
71
0.14
35
0.14
36
0.06
19
0.09
83
0.02
25
0.14
36
Gua
yam
a 2
0.06
85
0.07
47
0.11
11
0.10
63
0.07
22
0.09
09
0.07
22
0.10
63
Agu
as B
uena
s 0.
0222
0.
0174
0.
0556
0.
0440
0.
0444
0.
0444
0.
0444
0.
0440
Cay
ey 1
0.
0239
0.
0187
0.
0602
0.
0480
0.
0481
0.
0481
0.
0481
0.
0480
Cay
ey 2
0.
0248
0.
0194
0.
0625
0.
0500
0.
0498
0.
0498
0.
0498
0.
0500
Gur
abo
0.08
89
0.07
46
0.12
22
0.08
21
0.05
44
0.06
43
0.01
24
0.10
10
Car
olin
a 0.
0556
0.
0517
0.
0889
0.
0812
0.
0778
0.
0778
0.
0778
0.
0812
Mau
nabo
2
0.05
46
0.05
04
0.09
09
0.08
26
0.04
69
0.06
31
0.03
02
0.08
26
65
Tab
le 5
. Con
tinue
d.
Popu
latio
n C
abo
Roj
o M
oca
Isab
ela
May
agüe
z G
uáni
ca
Yau
co
Hat
illo
Are
cibo
Ju
ncos
0.
0124
0.
0174
0.
0363
0.
0440
0.
0149
0.
0348
0.
0149
0.
0440
Río
Gra
nde
0.02
49
0.04
02
0.07
78
0.06
87
0.05
72
0.06
67
0.06
67
0.06
87
Faja
rdo
0.04
60
0.05
17
0.08
89
0.08
12
0.06
85
0.07
78
0.07
78
0.08
12
Vie
ques
0.
0778
0.
0747
0.
1111
0.
1063
0.
1000
0.
1000
0.
1000
0.
1063
Cul
ebra
0.
1000
0.
0978
0.
1333
0.
1318
0.
1222
0.
1222
0.
1222
0.
1318
Sain
t Tho
mas
0.
2724
0.
2805
0.
3053
0.
3390
0.
2944
0.
2944
0.
2944
0.
3390
Sain
t Cro
ix 1
0.
4158
0.
4381
0.
4521
0.
5500
0.
4399
0.
4399
0.
4399
0.
5500
Sain
t Cro
ix 2
0.
4424
0.
4667
0.
4753
0.
5642
0.
4643
0.
4643
0.
4643
0.
5642
Sain
t Joh
n 0.
1778
0.
1795
0.
2111
0.
2235
0.
2000
0.
2000
0.
2000
0.
2235
Jost
Van
Dyk
e 0.
3748
0.
3929
0.
4092
0.
4815
0.
3977
0.
3977
0.
3977
0.
4815
Torto
la 2
0.
2018
0.
2051
0.
2379
0.
2584
0.
2258
0.
2258
0.
2258
0.
2584
66
Tab
le 5
. Con
tinue
d.
Popu
latio
n Pe
ñuel
as
Juan
a D
íaz
1 C
iale
s Ju
ana
Día
z 2
Man
atí
Dor
ado
Aib
onito
To
a A
lta
C
abo
Roj
o —
—
—
—
—
—
—
—
Moc
a —
—
—
—
—
—
—
—
Isab
ela
—
—
—
—
—
—
—
—
May
agüe
z —
—
—
—
—
—
—
—
Guá
nica
—
—
—
—
—
—
—
—
Yau
co
—
—
—
—
—
—
—
—
Hat
illo
—
—
—
—
—
—
—
—
Are
cibo
—
—
—
—
—
—
—
—
Peñu
elas
0.
0 —
—
—
—
—
—
—
Juan
a D
íaz
1 0.
2761
0.
0 —
—
—
—
—
—
Cia
les
0.35
97
0.02
09
0.0
—
—
—
—
—
Juan
a D
íaz
2 0.
1227
0.
0266
0.
0729
0.
0 —
—
—
—
67
Tab
le 5
. Con
tinue
d.
Popu
latio
n Pe
ñuel
as
Juan
a D
íaz
1 C
iale
s Ju
ana
Día
z 2
Man
atí
Dor
ado
Aib
onito
To
a A
lta
D
orad
o 0.
4092
0.
1260
0.
0775
0.
1222
0.
0663
0.
0 —
—
Aib
onito
0.
4331
0.
1408
0.
0555
0.
0959
0.
0886
0.
1248
0.
0 —
Toa
Alta
0.
2476
0.
0213
0.
0450
0.
0097
0.
0201
-0
.002
4 0.
0596
0.
0
Gua
yam
a 1
0.20
27
0.06
05
0.10
78
0.00
63
0.12
99
0.14
35
0.13
43
0.03
73
Gua
yam
a 2
0.35
44
0.10
03
0.07
75
0.07
60
0.09
75
0.11
11
0.08
41
0.05
90
Agu
as B
uena
s 0.
3520
0.
0674
0.
0232
0.
0667
0.
0411
0.
0556
0.
0684
0.
0124
Cay
ey 1
0.
4106
0.
0736
0.
0249
0.
0725
0.
0445
0.
0602
0.
0568
0.
0073
Cay
ey 2
0.
4370
0.
0767
0.
0258
0.
0752
0.
0461
0.
0625
0.
0561
0.
0248
Gur
abo
0.22
27
0.05
70
0.07
29
0.00
38
0.09
87
0.12
22
0.10
63
0.03
07
Car
olin
a 0.
3863
0.
1024
0.
0558
0.
1000
0.
0749
0.
0889
0.
1022
0.
0556
Mau
nabo
2
0.34
20
0.06
12
0.05
50
0.03
86
0.07
56
0.09
09
0.07
10
0.02
21
68
Tab
le 5
. Con
tinue
d.
Popu
latio
n Pe
ñuel
as
Juan
a D
íaz
1 C
iale
s Ju
ana
Día
z 2
Man
atí
Dor
ado
Aib
onito
To
a A
lta
Ju
ncos
0.
2923
0.
0403
0.
0232
0.
0175
0.
0411
0.
0556
0.
0364
0.
0023
Faja
rdo
0.38
63
0.10
24
0.05
58
0.09
09
0.07
49
0.08
89
0.04
95
0.05
56
Vie
ques
0.
4092
0.
1260
0.
0775
0.
1222
0.
0975
0.
1111
0.
1248
0.
0778
Cul
ebra
0.
4322
0.
1496
0.
0991
0.
1444
0.
1201
0.
1333
0.
1474
0.
1000
Sain
t Tho
mas
0.
6048
0.
3380
0.
2641
0.
3163
0.
2969
0.
3053
0.
3238
0.
2724
Sain
t Cro
ix 1
0.
8446
0.
5143
0.
3983
0.
4643
0.
4488
0.
4521
0.
4791
0.
4158
Sain
t Cro
ix 2
0.
7891
0.
5343
0.
4223
0.
4862
0.
4742
0.
4753
0.
5011
0.
4424
Sain
t Joh
n 0.
5136
0.
2339
0.
1741
0.
2222
0.
1996
0.
2111
0.
2269
0.
1778
Jost
Van
Dyk
e 0.
7500
0.
4603
0.
3597
0.
4207
0.
4047
0.
4092
0.
4331
0.
3748
Torto
la 2
0.
5956
0.
2659
0.
1970
0.
2500
0.
2261
0.
2379
0.
2559
0.
2018
69
Tab
le 5
. Con
tinue
d.
Popu
latio
n G
uaya
ma
1 G
uaya
ma
2 A
guas
Bue
nas
Cay
ey 1
C
ayey
2
Gur
abo
Car
olin
a M
auna
bo 2
C
abo
Roj
o —
—
—
—
—
—
—
—
Moc
a —
—
—
—
—
—
—
—
Isab
ela
—
—
—
—
—
—
—
—
May
agüe
z —
—
—
—
—
—
—
—
Guá
nica
—
—
—
—
—
—
—
—
Yau
co
—
—
—
—
—
—
—
—
Hat
illo
—
—
—
—
—
—
—
—
Are
cibo
—
—
—
—
—
—
—
—
Peñu
elas
—
—
—
—
—
—
—
—
Juan
a D
íaz
1 —
—
—
—
—
—
—
—
Cia
les
—
—
—
—
—
—
—
—
Juan
a D
íaz
2 —
—
—
—
—
—
—
—
70
Tab
le 5
. Con
tinue
d.
Popu
latio
n G
uaya
ma
1 G
uaya
ma
2 A
guas
Bue
nas
Cay
ey 1
C
ayey
2
Gur
abo
Car
olin
a M
auna
bo 2
M
anat
í —
—
—
—
—
—
—
—
Dor
ado
—
—
—
—
—
—
—
—
Aib
onito
—
—
—
—
—
—
—
—
Toa
Alta
—
—
—
—
—
—
—
—
Gua
yam
a 1
0.0
—
—
—
—
—
—
—
Gua
yam
a 2
0.06
16
0.0
—
—
—
—
—
—
Agu
as B
uena
s 0.
0862
0.
0556
0.
0 —
—
—
—
—
Cay
ey 1
0.
0943
0.
0602
-0
.017
0 0.
0 —
—
—
—
Cay
ey 2
0.
0499
0.
0228
0.
0 0.
0 0.
0 —
—
—
Gur
abo
0.02
06
0.07
60
0.06
67
0.07
25
0.05
60
0.0
—
—
Car
olin
a 0.
1205
0.
0889
0.
0333
0.
0360
0.
0174
0.
1000
0.
0 —
Mau
nabo
2
-0.0
244
0.00
78
0.03
07
0.03
33
-0.0
345
0.02
09
0.06
67
0.0
71
Tab
le 5
. Con
tinue
d.
Popu
latio
n G
uaya
ma
1 G
uaya
ma
2 A
guas
Bue
nas
Cay
ey 1
C
ayey
2
Gur
abo
Car
olin
a M
auna
bo 2
Ju
ncos
0.
0258
0.
0162
0.
0 0.
0 -0
.041
7 0.
0278
0.
0236
-0
.019
9
Faja
rdo
0.04
96
0.02
03
0.03
33
0.03
60
-0.0
248
0.07
22
0.05
72
-0.0
370
Vie
ques
0.
1435
0.
1111
0.
0556
0.
0602
0.
0228
0.
1222
0.
0703
0.
0909
Cul
ebra
0.
1666
0.
1333
0.
0778
0.
0848
0.
0881
0.
1444
0.
1111
0.
1154
Sain
t Tho
mas
0.
3480
0.
3053
0.
2505
0.
2828
0.
2969
0.
3163
0.
2834
0.
3127
Sain
t Cro
ix 1
0.
5153
0.
4521
0.
3919
0.
4643
0.
5000
0.
4643
0.
4278
0.
4988
Sain
t Cro
ix 2
0.
5339
0.
4753
0.
4204
0.
4931
0.
5237
0.
4862
0.
4534
0.
5224
Sain
t Joh
n 0.
2482
0.
2111
0.
1556
0.
1724
0.
1252
0.
2222
0.
1638
0.
2029
Jost
Van
Dyk
e 0.
4643
0.
4092
0.
3520
0.
4106
0.
3062
0.
4207
0.
3295
0.
4421
Torto
la 2
0.
2805
0.
2379
0.
1781
0.
2000
0.
0870
0.
2500
0.
1597
0.
2333
72
Tab
le 5
. Con
tinue
d.
Popu
latio
n Ju
ncos
R
io G
rand
e Fa
jard
o V
iequ
es
Cul
ebra
Sa
int T
hom
as
Sain
t Cro
ix 1
Cab
o R
ojo
—
—
—
—
—
—
—
Moc
a —
—
—
—
—
—
—
Isab
ela
—
—
—
—
—
—
—
May
agüe
z —
—
—
—
—
—
—
Guá
nica
—
—
—
—
—
—
—
Yau
co
—
—
—
—
—
—
—
Hat
illo
—
—
—
—
—
—
—
Are
cibo
—
—
—
—
—
—
—
Peñu
elas
—
—
—
—
—
—
—
Juan
a D
íaz
1 —
—
—
—
—
—
—
Cia
les
—
—
—
—
—
—
—
Juan
a D
íaz
2 —
—
—
—
—
—
—
73
Tab
le 5
. Con
tinue
d.
Popu
latio
n Ju
ncos
R
io G
rand
e Fa
jard
o V
iequ
es
Cul
ebra
Sa
int T
hom
as
Sain
t Cro
ix 1
Man
atí
—
—
—
—
—
—
—
Dor
ado
—
—
—
—
—
—
—
Aib
onito
—
—
—
—
—
—
—
Toa
Alta
—
—
—
—
—
—
—
Gua
yam
a 1
—
—
—
—
—
—
—
Gua
yam
a 2
—
—
—
—
—
—
—
Agu
as B
uena
s —
—
—
—
—
—
—
Cay
ey 1
—
—
—
—
—
—
—
Cay
ey 2
—
—
—
—
—
—
—
Gur
abo
—
—
—
—
—
—
—
Car
olin
a —
—
—
—
—
—
—
Mau
nabo
2
—
—
—
—
—
—
—
74
Tab
le 5
. Con
tinue
d.
Popu
latio
n Ju
ncos
R
io G
rand
e Fa
jard
o V
iequ
es
Cul
ebra
Sa
int T
hom
as
Sain
t Cro
ix 1
Junc
os
0.0
—
—
—
—
—
—
Río
Gra
nde
0.00
23
0.0
—
—
—
—
—
Faja
rdo
-0.0
069
0.05
56
0.0
—
—
—
—
Vie
ques
0.
0363
0.
0394
0.
0889
0.
0 —
—
—
Cul
ebra
0.
0778
0.
1000
0.
1111
0.
1333
0.
0 —
—
Sain
t Tho
mas
0.
2505
0.
2724
0.
2834
0.
3053
0.
3272
0.
0 —
Sain
t Cro
ix 1
0.
3919
0.
4158
0.
4278
0.
4521
0.
4766
0.
1791
0.
0
Sain
t Cro
ix 2
0.
4204
0.
4424
0.
4534
0.
4753
0.
4971
0.
1804
-0
.089
1
Sain
t Joh
n 0.
1294
0.
1253
0.
1889
0.
1608
0.
2333
0.
4034
0.
5646
Jost
Van
Dyk
e 0.
2923
0.
2476
0.
3863
0.
2885
0.
4322
0.
6048
0.
8446
Torto
la 2
0.
1220
0.
0842
0.
2138
0.
1247
0.
2622
0.
4543
0.
6739
75
Tab
le 5
. Con
tinue
d.
Popu
latio
n Sa
int C
roix
2
Sain
t Joh
n Jo
st V
an D
yke
Torto
la 2
C
abo
Roj
o —
—
—
—
Moc
a —
—
—
—
Isab
ela
—
—
—
—
May
agüe
z —
—
—
—
Guá
nica
—
—
—
—
Yau
co
—
—
—
—
Hat
illo
—
—
—
—
Are
cibo
—
—
—
—
Peñu
elas
—
—
—
—
Juan
a D
íaz
1 —
—
—
—
Cia
les
—
—
—
—
Juan
a D
íaz
2 —
—
—
—
76
Tab
le 5
. Con
tinue
d.
Popu
latio
n Sa
int C
roix
2
Sain
t Joh
n Jo
st V
an D
yke
Torto
la 2
M
anat
í —
—
—
—
Dor
ado
—
—
—
—
Aib
onito
—
—
—
—
Toa
Alta
—
—
—
—
Gua
yam
a 1
—
—
—
—
Gua
yam
a 2
—
—
—
—
Agu
as B
uena
s —
—
—
—
Cay
ey 1
—
—
—
—
Cay
ey 2
—
—
—
—
Gur
abo
—
—
—
—
Car
olin
a —
—
—
—
Mau
nabo
2
—
—
—
—
77
Tab
le 5
. Con
tinue
d.
Popu
latio
n Sa
int C
roix
2
Sain
t Joh
n Jo
st V
an D
yke
Torto
la 2
Ju
ncos
—
—
—
—
Río
Gra
nde
—
—
—
—
Faja
rdo
—
—
—
—
Vie
ques
—
—
—
—
Cul
ebra
—
—
—
—
Sain
t Tho
mas
—
—
—
—
Sain
t Cro
ix 1
—
—
—
—
Sain
t Cro
ix 2
0.
0 —
—
—
Sain
t Joh
n 0.
5733
0.
0 —
—
Jost
Van
Dyk
e 0.
7891
0.
3454
0.
0 —
Torto
la 2
0.
6609
0.
1882
-0
.001
1 0.
0
78
Table 6. Island, population number, municipality, number of samples, heterozygosity, and
number of loci calculated from 16,225 SNPs yielded from double digest restriction-site
associated DNA sequencing of 114 individuals of Leptodactylus albilabris.
Island Population number Municipality Number of
samples Heterozygosity Number of loci
Puerto Rico 1 Cabo Rojo 4 0.1778 +/- 0.0021 11311
Puerto Rico 2 Moca 3 0.1737 +/- 0.0021 14700
Puerto Rico 3 Isabela 4 0.174 +/- 0.002 14118
Puerto Rico 4 Mayagüez 2 0.206 +/- 0.0027 12951
Puerto Rico 6 Yauco 5 0.1793 +/- 0.0018 13206
Puerto Rico 10 Juana Díaz 5 0.1818 +/- 0.0018 13042
Puerto Rico 11 Ciales 4 0.2016 +/- 0.0021 12564
Puerto Rico 13 Manatí 4 0.2023 +/- 0.002 14590
Puerto Rico 14 Dorado 5 0.1894 +/- 0.0019 14419
Puerto Rico 15 Aibonito 3 0.2196 +/- 0.0023 14643
Puerto Rico 19 Guayama 5 0.1914 +/- 0.0018 14413
Puerto Rico 20 Aguas Buenas 4 0.2325 +/- 0.0023 12343
Puerto Rico 24 Carolina 5 0.1786 +/- 0.0018 13483
Puerto Rico 28 Río Grande 5 0.1919 +/- 0.0018 14689
Puerto Rico 29 Fajardo 5 0.2030 +/- 0.0018 14241
79
Table 6. Continued.
Island Population number Municipality Number of
samples Heterozygosity Number of loci
Vieques 1 30 — 5 0.1722 +/- 0.0018 14069
Culebra 32 — 5 0.1214 +/- 0.0017 14003
Saint Thomas 1 33 — 7 0.1085 +/- 0.0016 14129
Saint Croix 2 36 — 7 0.0979 +/- 0.0015 14112
Saint John 38 — 10 0.1039 +/- 0.0015 13637
Jost Van Dyke 1 39 — 11 0.0993 +/- 0.0015 14140
Tortola 2 42 — 6 0.1326 +/- 0.0017 14752
80
Figure 1. Map of the major islands of the Puerto Rican Bank, in the eastern Caribbean Sea.
Orange circles indicate the approximate geographic locations of the sampling localities of
Leptodactylus albilabris. The approximate land configuration at maximum sea level lowering
(ca. –134 m) during the Last Glacial Maximum (ca. 29,000 – 21,000 ya; Lambeck et al., 2014) is
shown in grey. Note the changes in area, isolation, and connectivity of islands. AN = Anegada,
CU = Culebra, JVD = Jost Van Dyke, PR = Puerto Rico, STC = Saint Croix, STJ = Saint John,
STT = Saint Thomas, TO = Tortola, VG = Virgin Gorda, VI = Vieques.
81
82
Figure 2. Bayesian Inference tree (generated by MrBayes 3.2.6; Ronquist et al., 2012) estimated
for 140 unique cytochrome b haplotypes of Leptodactylus albilabris, using L. poecilochilus as
the outgroup. Numbers at nodes designated by black circles are ≥ 90% posterior probability
values. Colored (green, yellow, blue, red, grey) branches represent the physiographic region(s) of
Puerto Rico where the haplotype occurs; white branches indicate that the haplotype is present in
one or more Eastern Island (CU = Culebra, VI = Vieques, TO = Tortola, STT = Saint Thomas,
STC = Saint Croix, STJ = Saint John, JVD = Jost Van Dyke). Taxon labels consist of the
population number(s) where a particular haplotype was detected, followed by the specific
haplotype number (preceded by the letter H) in parentheses. Asterisks indicate the two most
inclusive, well-supported phylogroups. The light grey box indicates the clade consisting of
Vieques 1, 2 (Populations 30, 31, respectively). PG = phylogroup.
83
84
Figure 2. Continued.
85
Figure 3. Maximum parsimony network (estimated with NETWORK 5.0.0.3; Bandelt et al.,
1999) representing the relationships among 140 unique cytochrome b haplotypes recovered from
322 samples of Leptodactylus albilabris. Colors indicate the island or region where the
haplotypes occur (i.e. Puerto Rico, Vieques, Culebra, Saint Thomas, Saint John, Saint Croix, Jost
Van Dyke, Tortola). Numbered circles represent unique haplotypes; hatch marks indicate single
mutations; black squares indicate missing (i.e. unsampled or extinct) haplotypes. Circle size is
proportional to haplotype frequency. The three haplotypes with an inner circle (i.e. 1, 88; 17, 73;
28, 108, 121) represent merged haplotypes, for NETWORK recognizes these haplotypes as
equivalent. Specifically, haplotypes 1 and 88 and haplotypes 17 and 73 differ by a single,
unspecified base pair (N), whereas haplotypes 28, 108, and 121 differ by three unspecified base
pairs.
86
87
Figure 4. Genetic clustering of 16,140 single nucleotide polymorphic loci estimated by the R
package LEA (Frichot and François, 2015). The five genetically distinct groups are color coded
(light grey, yellow, blue, red, dark grey). Pie charts indicate the approximate geographic
locations of the sampling localities, and illustrate the proportion of each genetic group in a given
population. Sparse non-negative matrix factorization (snmf) analysis shows the assignment of
each individual sample in a population to a particular genetic group. Populations are listed below
the bar plots, from west to east. PR = Puerto Rico, VI = Vieques, CU = Culebra, STT = Saint
Thomas, STJ = Saint John, STC = Saint Croix, JVD = Jost Van Dyke, TO = Tortola.
88
89
Figure 5. Approximate configurations of the Puerto Rican Bank (PRB) at six different sea levels
(-10 m – -60 m) during the Last Glacial Maximum (ca. 29,000 – 21,000 ya; Lambeck et al.,
2014). Bathymetric data were obtained from the General Bathymetric Chart of the Oceans
https://www.gebco.net/.
A) 0 m. Present-day sea levels, illustrating the separation of the largest islands of the PRB.
B) -10 m. Saint John, Jost Van Dyke, Tortola, Virgin Gorda, and Anegada are connected by a
land bridge.
C) -20 m. Puerto Rico and Vieques are now connected by a land bridge; Saint Thomas is now
partially connected to Saint John by a land bridge.
D) -30 m. Puerto Rico, Vieques, and Culebra, are now connected by a land bridge; Saint Thomas
is now completely connected to Saint John, Jost Van Dyke, Tortola, Virgin Gorda, and Anegada
by a more extensive land bridge.
E) -40 m. All the islands of the PRB are now connected by a land bridge.
F) -50 m. The PRB has slightly greater subaerial exposure.
G) -60 m. The configuration and exposure of the PRB are very similar to those during maximum
sea level lowering (-134 m; Figure 1), due to the steep slopes of the PRB (Heatwole &
MacKenzie, 1967). Note that Saint Croix is never connected to the PRB.
AN = Anegada, CU = Culebra, JVD = Jost Van Dyke, PR = Puerto Rico, STC = Saint Croix,
STJ = Saint John, STT = Saint Thomas, TO = Tortola, VG = Virgin Gorda, VI = Vieques.
90
91
Figure 5. Continued.
92
References
Balinsky, J. B. 1981. Adaptation of nitrogen metabolism to hyperosmotic environment in
Amphibia. Journal of Experimental Zoology 215:335–350.
Bandelt, H. J., Forster, P., and A. Röhl. 1999. Median-joining networks for inferring intraspecific
phylogenies. Molecular Biology and Evolution 16:37–48.
Baptestini, E. M., de Aguiar, M. A. M., and Y. Bar–Yam. 2013. Conditions for neutral speciation
via isolation by distance. Journal of Theoretical Biology 335:51–56.
Barbour, T. 1930. Some faunistic changes in the Lesser Antilles. Proceedings of the New
England Zoologists’ Club 11:73-85.
Barker, B. S., and J. A. Rodríguez-Robles. 2017. Origins and genetic diversity of introduced
populations of the Puerto Rican Red-eyed Coquí, Eleutherodactylus antillensis, in Saint
Croix (U.S. Virgin Islands) and Panamá. Copeia 105:220–228.
Barker, B. S., Rodríguez-Robles, J. A., Aran, V. S., Montoya, A., Waide, R. B., and J. A. Cook.
2012. Sea level, topography, and island diversity: phylogeography of the Puerto Rican
Red-eyed Coquí, Eleutherodactylus antillensis. Molecular Ecology 21:6033–6052.
Barrow, L. N., Soto-Centeno, J. A., Warwick, A. R., Lemmon, A. R., and E. M. Lemmon. 2017.
Evaluating hypotheses of expansion from refugia through comparative phylogeography
of south-eastern Coastal Plain amphibians. Journal of Biogeography 44:2692–2705.
Bell, R. C., Drewes, R. C., Channing, A., Gvoždík, V., Kielgast, J., Lötters, S., Stuart, B. L., and
K. R. Zamudio. 2015. Overseas dispersal of Hyperolius reed frogs from Central Africa to
the oceanic islands of São Tomé and Príncipe. Journal of Biogeography 42:65–75.
Bintanja, R., van de Wal, R. S. W., and J. Oerlemans. 2005. Modelled atmospheric temperatures
and global sea levels over the past million years. Nature 437:125–128.
93
Calsbeek, R., and T. B. Smith. 2003. Ocean currents mediate evolution in island lizards. Nature
426:552–555.
Censky, E. J. 1989. Eleutherodactylus johnstonei (Salientia: Leptodactylidae) from Anguilla,
West Indies. Caribbean Journal of Science 25:229-230.
Censky, E. J., Hodge, K., and J. Dudley. 1998. Over-water dispersal of lizards due to hurricanes.
Nature 395:556.
Clouse, R. M., Janda, M., Blanchard, B., Sharma, P., Hoffman, B. D., Andersen,
A.N., Czekanski Moir, J. E., Krushelnycky, P., Rabeling, C., Wilson, E. O., Economo, E.
P., Sarnat, E. M., General, D. M., Alpert, G. D., and W. C. Wheeler. 2015. Molecular
phylogeny of Indo Pacific carpenter ants (Hymenoptera: Formicidae, Camponotus)
reveals waves of dispersal and colonization from diverse source areas. Cladistics 31:424–
437.
Cochran, D. M. 1923. A new frog of the genus Leptodactylus. Journal of the Washington
Academy of Sciences 13:184–185.
Cochran, D. M. 1941. The herpetology of Hispaniola. Smithsonian Institution, Bulletin of the
United States National Museum 177:1–398.
da Fonesca, R. R., Albrechtsen, A., Themudo, G. E., Ramos-Madrigal, J., Sibbesen, J. A.,
Maretty, L., Zepeda-Mendoza, M. L., Campos, P. F., Heller, R., and R. J. Pereira. 2016.
Next-generation biology: Sequencing and data analysis approaches for non-model
organisms. Marine Genomics 30:3–13.
Darriba, D., Taboada, G. L., Doallo, R., and D. Posada. 2012. jModelTest 2: more models, new
heuristics and parallel computing. Nature Methods 9:772.
94
Davison, A., and S. Chiba. 2008. Contrasting response to Pleistocene climate change by ground-
living and arboreal Mandarina snails from the oceanic Hahajima archipelago.
Philosophical Transactions of the Royal Society B 363:3391–3400.
Deli, T., Kiel, C., and C. D. Schubart. 2019. Phylogeographic and evolutionary history analyses
of the warty crab Eriphia verrucosa (Decapoda, Brachyura, Eriphiidae) unveil genetic
imprints of a late Pleistocene vicariant event across the Gibraltar Strait, erased by
postglacial expansion and admixture among refugial lineages. BMC Evolutionary
Biology 19: https://doi.org/10.1186/s12862-019-1423-2.
de Aguiar, M. A. M., Baranger, M., Baptestini, E. M., Kaufman, L., and Y. Bar–Yam. 2009.
Global patterns of speciation and diversity. Nature 460:384–387.
de Sá, R. O., Grant, T., Camargo, A., Heyer, W. R., Ponssa, M. L., and E. Stanley. 2014.
Systematics of the neotropical genus Leptodactylus Fitzinger, 1826 (Anura:
Leptodactyilidae): phylogeny, the relevance of non-molecular evidence, and species
accounts. South American Journal of Herpetology 9:1–128.
Dent, J. N. 1956. Observations on the life history and development of Leptodactylus albilabris.
Copeia 4:207–210.
Duellman, W. E., and L. Trueb. 1994. Biology of Amphibians. The John Hopkins University
Press, Baltimore, Maryland.
Duryea, M. C., Zamudio, K. R., and C. A. Brasileiro. 2015. Vicariance and marine migration in
continental island populations of a frog endemic to the Atlantic Coastal forest. Heredity
115:225–234.
95
Ewel, J. J., and J. L. Whitmore. 1973. The Ecological Life Zones of Puerto Rico and the U.S.
Virgin Islands. USDA Forest Service, Institute of Tropical Forestry, Research Paper:1–
72.
Excoffier, L., and H. E. L. Lischer. 2010. Arlequin Suite ver 3.5, a new series of programs to
perform population genetics analyses under Linux and Windows. Molecular Ecology
Resources 10:564–567.
Ezer, T. 2019. Numerical modeling of the impact of hurricanes on ocean dynamics: sensitivity of
the Gulf Stream response to storm’s track. Ocean Dynamics 69:1053–1066.
Falk, B. G., and S. L. Perkins. 2013. Host specificity shapes population structure of pinworm
parasites in Caribbean reptiles. Molecular Ecology 22:4576
Fernández-Palacios, J. M., Rijsdijk, K. F., Norder, S. J., Otto, R., de Nascimento, L., Fernández-
Lugo, S., Tjørve, E., and R. J. Whittaker. 2016. Towards a glacial-sensitive model of
island biogeography. Global Ecology and Biogeography 25:817–830.
Flores-Nieves, C. Z., Logue, D. M., and C. J. Santos-Flores. 2014. Native White-lipped frog
larvae (Leptodactylus albilabris) outcompete introduced Cane toad larvae (Rhinella
marina) under laboratory conditions. Herpetological Conservation and Biology 9:378–
386.
Frankham, R. 1997. Do island populations have less genetic variation than mainland
populations? Heredity 78:311–327.
Frichot, E., and O. François. 2015. LEA: An R package for landscape and ecological association
studies. Methods in Ecology and Evolution 6:925–929.
Frichot, E., Mathieu, F., Trouillon, T., Bouchard, G., and O. François. 2014. Fast and efficient
estimation of individual ancestry coefficients. Genetics 196:973–983.
96
Furlan, E., Stoklosa, J., Griffiths, J., Gust, N., Ellis, R., Huggins, R. M., and A. R. Weeks. Small
population size and extremely low levels of genetic diversity in island populations of the
platypus, Ornithorhynchus anatinus. Ecology and Evolution 2:844–857.
Garcia-Porta, J., Litvinchuk, S. N., Crochet, P. A., Romano, A., Geniez, P. H., Lo-Valvo, M.,
Lymberakis, P., and S. Carranza. 2012. Molecular phylogenetics and historical
biogeography of the west-palearctic common toads (Bufo bufo species complex).
Molecular Phylogenetics and Evolution 63:113–130.
Garg, K. M., Chattopadhyay, B., Wilton, P. R., Prawiradilaga, D. M., and F. E. Rheindt. 2018.
Pleistocene land bridges act as semipermeable agents of avian gene flow in Wallacea.
Molecular Phylogenetics and Evolution 125:196–203.
Gehara, M., Garda, A. A., Werneck, F. P., Oliviera, E. F., da Fonseca, E. M., Camurugi, F.,
Magalhães, F. M., Lanna, F. M., Sites, J. W., Marques, R., Silveira-Filho, R., São Pedro,
V. A., Colli, G. R., Costa, G. C., and F. T. Burbrink. 2017. Estimating synchronous
demographic changes across populations using hABC and its application for a
herpetological community from northeastern Brazil. Molecular Ecology 26:4756–4771.
Gill, I. P., Hubbard, D. K., McLaughlin, P., and C. Moore. 1989. Sedimentological and tectonic
evolution of Tertiary St. Croix, pp. 49–71. In: Hubbard, D. K. (ed.), 12th Caribbean
Geological Conference, West Indies Laboratory. Teague Bay, Saint Croix.
Gómez-Mestre, I., and M. Tejedo. 2003. Local adaptation of an anuran amphibian to osmotically
stressful environments. Evolution 57:1889–1899.
Goudet, J. 2005. HIERFSTAT, a package for R to compute and test hierarchical F-statistics.
Molecular Ecology Notes 5:184–186.
97
Guindon, S., and O. Gascuel. 2003. A simple, fast and accurate method to estimate large
phylogenies by maximum-likelihood. Systematic Biology 52:696–704.
Hassine, J. B., Gutiérrez-Rodríguez, J., Escoriza, D., and I. Martínez-Solano. 2016. Inferring the
roles of vicariance, climate and topography in population differentiation in Salamandra
algira (Caudata, Salamandridae). Journal of Zoological Systematics and Evolutionary
Research 54:116–126.
Heatwole, H., De Austin, S. B., and R. Herrero. 1968. Heat tolerances of tadpoles of two species
of tropical anurans. Comparative Biochemistry and Physiology 27:807–815.
Heatwole, H., and R. Levins. 1972. Biogeography of the Puerto Rican Bank: flotsam transport of
terrestrial animals. Ecology 53:112–117.
Heatwole, H., and F. Mackenzie. 1967. Herpetogeography of Puerto Rico. IV. Paleogeography,
faunal similarity and endemism. Evolution 21:429–438.
Hedges, S. B. 1999. Distribution Patterns of Amphibians in the West Indies, pp. 211–254. In:
Duellman, W. E. (ed.), Regional Patterns of Amphibian Distribution: a Global
Perspective. Johns Hopkins University Press, Baltimore, Maryland.
Hedges, S. B., Hass, C. A., and L. R. Maxson. 1992. Caribbean biogeography: molecular
evidence for dispersal in West Indian terrestrial vertebrates. Proceedings of the National
Academy of Sciences, USA 89:1909–1913.
Hedges, S. B. and M. P. Heinicke. 2007. Molecular phylogeny and biogeography of West Indian
frogs of the genus Leptodactylus (Anura, Leptodactylidae). Molecular Phylogenetics and
Evolution 44:308–314.
98
Heinicke, M. P., Duellman, W. E., and S. B. Hedges. 2007. Major Caribbean and Central
American frog faunas originated by ancient oceanic dispersal. Proceedings of the
National Academy of Sciences, USA 104:10092–10097.
Helmer, E. H., Ramos, O., López T. D. M., Quiñones, M., and W. Diaz. 2002. Mapping the
forest type and land cover of Puerto Rico, a component of the Caribbean biodiversity
hotspot. Caribbean Journal of Science 38:165–183.
Henderson, R. W., and S. B. Hedges. 1995. Origin of West Indian populations of the
geographically widespread Boa Corallus enhydris inferred from mitochondrial DNA
sequences. Molecular Phylogenetics and Evolution 4:88–92.
Henderson, R. W., and R. Powell. 2009. Natural History of West Indian Reptiles and
Amphibians. University Press of Florida, Florida
Hewitt, G. M. 1996. Some genetic consequences of ice ages, and their role in divergence and
speciation. Biological Journal of the Linnean Society 58:247–276.
Hewitt, G. M. 2000. The genetic legacy of the Quaternary ice ages. Nature 405:907–913.
Hewitt, G. M. 2004. Genetic consequences of climatic oscillations in the Quaternary.
Philosophical Transactions of the Royal Society of London B-Biological Sciences
359:183–195.
Heyer, W. R. 1978. Systematics of the fuscus group of the frog genus Leptodactylus (Amphibia,
Leptodactylidae). Science Bulletin, Natural History Museum of Los Angeles County
29:1–85.
Hirano, T., Kameda, Y., Saito, T., and S. Chiba. 2019. Divergence before and after the isolation
of islands: Phylogeography of the Bradybaena land snails on the Ryukyu Islands of
Japan. Journal of Biogeography 46:1197–1213.
99
Hoffman, E. A., and M. S. Blouin. 2004. Evolutionary history of the Northern Leopard frog:
reconstruction of phylogeny, phylogeography, and historical changes in population
demography from mitochondrial DNA. Evolution 58:145–159.
Hohenlohe, P. A., Bassham, S., Etter, P. D., Stiffler, N., Johnson, E. A., and W. A. Cresko.
2010. Population genomics of parallel adaptation in threespine stickleback using
sequenced RAD tags. PLoS Genetics 6:e1000862
https://doi.org/10.1371/journal.pgen.1000862.
Hyseni, C., and R. C. Garrick. 2019. The role of glacial interglacial climate change in shaping
the genetic structure of eastern subterranean termites in the southern Appalachian
Mountains, USA. Ecology and Evolution 9:4621–4636.
Joglar, R. 2005. Anfibios, pp. 39–96. In: Joglar, R. (ed.), Biodiversidad de Puerto Rico:
Vertebrados Terrestres y Ecosistemas. Instituto de Cultura Puertorriqueña. San Juan,
Puerto Rico.
Jukes, T. H., and C. R. Cantor. 1969. Evolution of Protein Molecules, pp. 21–132. In: Munro, H.
N. (ed.), Mammalian Protein Metabolism. Academic Press, New York.
Kaiser, H., Barrio-Amorós, C. L., Trujillo, J. D., and J. D. Lynch. 2002. Expansion of
Eleutherodactylus johnstonei in northern South America: Rapid dispersal through human
interactions. Herpetological Review 33:290-294.
Kidd, D. M., and M. G. Ritchie. 2006. Phylogeographic information systems: putting the
geography into phylogeography. Journal of Biogeography 33:1851–1865.
Kimura, M. 1980. A simple method for estimating evolutionary rates of base substitutions
through comparative studies of nucleotide sequences. Journal of Molecular Evolution
16:111–120.
100
Kimura, M., and G. H. Weiss. 1964. The stepping stone model of population structure and the
decrease of genetic correlation with distance. Genetics 49:561–576.
Krysko, K. L., Nuñez, L. P., Lippi, C. A., Smith, D. J., and M. C. Granatosky. 2016. Pliocene–
Pleistocene lineage diversifications in the Eastern Indigo Snake (Drymarchon couperi) in
the southeastern United States. Molecular Phylogenetics and Evolution 98:111–122.
Lambeck, K., Rouby, H., Purcell, A., Sun, Y., and M. Sambridge. 2014. Sea level and global ice
volumes from the Last Glacial Maximum to the Holocene. Proceedings of the National
Academy of Sciences, USA 111:15296–15303.
Leal, B. S. S., da Silva, C. P., and F. Pinheiro. 2016. Phylogeographic studies depict the role of
space and time scales of plant speciation in a highly diverse neotropical region. Critical
Reviews in Plant Sciences 35:215–230.
Lever, C. 2003. Naturalized reptiles and amphibians of the world. Oxford University Press.
Oxford, United Kingdom.
López, P. T., Narins, P. M., Lewis, E. R., and S. W. Moore. 1988. Acoustically induced call
modification in the White-lipped Frog, Leptodactylus albilabris. Animal Behavior
36:1295–1308.
Losos, J. B. 2011. Convergence, adaptation, and constraint. Evolution 65:1827–1840.
Losos, J. B. and R. E. Ricklefs. 2009. Adaptation and diversification on islands.
Nature 457:830–836.
Luu, K., Bazin, E., and M. G. B. Blum. 2016. pcadapt: an R package to perform genome scans
for selection based on principle component analysis. Molecular Ecology Resources
17:67–77.
101
MacArthur, R. H., and E. O. Wilson. 1967. The Theory of Island Biogeography. Princeton
University Press. Princeton, New Jersey.
MacLean, W. P. 1982. Reptiles and amphibians of the Virgin Islands. Macmillan Education Ltd.,
London.
Messer, P. W., and D. A. Petrov. 2013. Population genomics of rapid evolution adaptation by
soft selective sweeps. Trends in Ecology & Evolution 28:659–669.
Miller, M. P. 2005. Alleles in Space (AIS): Computer software for the joint analysis of
interindividual spatial and genetic information. Journal of Heredity 96:722–724.
Morales, M. 2017. Sciplot: scientific graphing functions for factorial designs. The R
Development Core Team and with general advice from the R-help listserv community.
Moriyama, J., Takeuchi, H., Ogura-Katayama, A., and T. Hikida. 2018. Phylogeography of the
Japanese ratsnake, Elaphe climacophora (Serpentes: Colubridae): impacts of Pleistocene
climatic oscillations and sea-level fluctuations on geographical range. Biological Journal
of the Linnean Society 124:174–187.
Murphy, P. G., Lugo, A. E., Murphy, A. J., and D. C. Nepstad. 1995. The dry forests of Puerto
Rico’s south coast, pp. 178–209. In: Lugo, A. E., and C. Lowe (ed.), Tropical Forests:
Management and Ecology. Springer–Verlag, New York.
Nguiffo, D. N., Mpoame, M., and C. S. Wondji. 2019. Genetic diversity and population structure
of goliath frogs (Conraua goliath) from Cameroon. Mitochondrial DNA Part A 30:657–
663.
Nei, M., Maruyama, T., and R. Chakraborty. 1975. The bottleneck effect and genetic variability
in populations. Evolution 29:1–10.
102
Papadopoulou, A., and L. L. Knowles. 2015. Genomic tests of the species pump hypothesis:
Recent island connectivity cycles drive population divergence but not speciation in
Caribbean crickets across the Virgin Islands. Evolution 69:1501–1517.
Papadopoulou, A., and L. L. Knowles. 2017. Linking micro- and macroevolutionary perspectives
to evaluate the role of Quaternary sea-level oscillations in island diversification.
Evolution 71:2901–2917.
Parvizi, E., Naderloo, R., Keikhosravi, A., Solhjouy-Fard, S., and C. D. Schubart. 2018. Multiple
Pleistocene refugia and repeated phylogeographic breaks in the southern Caspian Sea
region: Insights from the freshwater crab Potamon ibericum. Journal of Biogeography
45:1234–1245.
Perry, G., Powell, R., and H. Watson. 2006. Keeping invasive species off Guana Island, British
Virgin Islands. Iguana: Conservation, Natural History, and Husbandry of Reptiles
13:273–277.
Petit, R. J., Aguinagalde, I., de Beaulieu, J., Bittkau, C., Brewer, S., Cheddadi, R., Ennos, R.,
Fineschi, S., Grivet, D., Lascoux, M., Mohanty, A., Müller-Starck, G., Demesure-Musch,
B., Palmé, A., Martín, J. P., Rendell, S., and G. G. Vendramin. 2003. Glacial refugia:
hotspots but not melting pots of genetic diversity. Science 300:1563–1565.
Platenberg, R. J. 2007. Impacts of Introduced Species on an Island Ecosystem: Non-native
Reptiles and Amphibians in the U.S. Virgin Islands, pp. 168–174. In: Witmer, G. W.,
Pitt, W. C., and K. A Fagerstone (eds.), Managing Vertebrate Invasive Species:
Proceedings of an International Symposium. USDA/APHIS/WS National Wildlife
Research Center. Fort Collins, Colorado.
103
Polzin. T., and S. V. Daneshmand. 2003. On Steiner trees and minimum spanning trees in
hypergraphs. Operations Research Letters 31:12–20.
Potter, S., Xue, A. T., Bragg, J. G., Rosauer, D. F., Roycroft, E. J., and C. Moritz. 2018.
Pleistocene climatic changes drive diversification across a tropical savanna. Molecular
Ecology 27:520–532.
Powell, R. 2006. Conservation of the herpetofauna on the Dutch Windward Islands: St.
Eustatius, Saba, and St. Maarten. Applied Herpetology 3:293–306.
Powell, R., Henderson, R. W., Farmer, M. C., Breuil, M., Echternacht, A. C., van Buurt, G.,
Romagosa, C. M., and G. Perry. 2011. Introduced amphibians and reptiles in the greater
Caribbean: patterns and conservation implications, pp. 63–144. In: Hailey A., Wilson,
B.S., and J. A. Horrocks. (eds.), Conservation of Caribbean Island Herpetofaunas
Volume 1: Conservation Biology and the Wider Caribbean. Brill, Boston.
Powell, R., Henderson, R.W., and J. S. Parmerlee, Jr. 2005. Reptiles and Amphibians of the
Dutch Caribbean: St. Eustatius, Saba, and St. Maarten. St. Eustatius National Parks
Foundation, Gallows Bay, St. Eustatius, Netherlands Antilles.
Pregill, G. K., and S. L. Olson. 1981. Zoogeography of West Indian vertebrates in relation to
Pleistocene climatic cycles. Annual Review of Ecology and Systematics 12:75–98.
Provan, J., and K. D. Bennett. 2008. Phylogeographic insights into cryptic glacial refugia. Trends
in Ecology & Evolution 23:564–571.
Puritz, J. B., Hollenbeck, C. M., and J. R. Gold. 2014a. dDocent: a RADseq, variant-calling
pipeline designed for population genomics of non-model organisms. PeerJ 2:e431
https://doi.org/10.7717/peerj.431.
104
Puritz, J. B., Matz, M. V., Toonen, R. J., Weber, J. N., Bolnick, D. I., and C. E. Bird. 2014b.
Demystifying the RAD fad. Molecular Ecology 23:5937–5942.
Rambaut, A. and A. J., Drummond. 2010. FigTree v1.3.1. Institute of Evolutionary Biology,
University of Edinburgh, Edinburgh. http://tree.bio.ed.ac.uk/software/figtree/.
Reilly, S. B., Stubbs, A. L., Karin, B. R., Arida, E., Iskandar, D. T., and J. A. McGuire. 2019.
Recent colonization and expansion through the Lesser Sundas by seven amphibian and
reptile species. Zoologica Scripta 48:614–626.
Renken, R. A., Ward, W. C., Gill, I. P., Gómez-Gómez, F., and J. Rodríguez-Martínez. 2002.
Geology and hydrology of the Caribbean islands aquifer system of the Commonwealth of
Puerto Rico and the U.S. Virgin Islands. Regional Aquifer-System Analysis—Caribbean
Islands. U.S. Geological Survey Professional Paper 1419, U.S. Department of the
Interior, Reston, Virginia.
Reynolds, R. G., Strickland, T. R., Kolbe, J. J., Falk, B. G., Perry, G., Revell, L. J., and J. B.
Losos. 2017. Archipelagic genetics in a widespread Caribbean anole. Journal of
Biogeography 44:2631–2647.
Ricklefs, R., and E. Bermingham. 2008. The West Indies as a laboratory of biogeography and
evolution. Philosophical Transactions of the Royal Society B-Biological Sciences
363:2393–2413.
Ricklefs, R. E. and I. J. Lovette. 1999. The roles of island area per se and habitat diversity in the
species-area relationships of four Lesser Antillean faunal groups. Journal of Animal
Ecology 68:1142–1160.
Rochette, N. C., and J. M. Catchen. 2017. Deriving genotypes from RAD-seq short-read data
using Stacks. Nature Protocols 12:2640–2659.
105
Rödder, D. 2009. Human footprint, facilitated jump dispersal, and the potential distribution of
the invasive Eleutherodactylus johnstonei Barbour 1914 (Anura Eleutherodactylidae).
Tropical Zoology 22:205–217.
Rodríguez-Robles, J. A., Jezkova, T., Fujita, M. K., Tolson, P. J., and M. A. García. 2015.
Genetic divergence and diversity in the Mona and Virgin Islands Boas, Chilabothrus
monensis (Epicrates monensis) (Serpentes: Boidae), West Indian snakes of special
conservation concern. Molecular Phylogenetics and Evolution 88:144–153.
Roesti, M., Salzburger, W., and D. Berner. 2012. Uninformative polymorphisms bias genome
scans for signatures of selection. BMC Evolutionary Biology 12:94.
Rohling, E. J., Fenton, M., Jorissen, F. J., Bertrand, P., Ganssen, G., and J. P. Caulet. 1998.
Magnitudes of sea-level lowstands of the past 500,000 years. Nature 394:162–165.
Ronquist, F., Teslenko, M., van der Mark, P., Ayres, D. L., Darling, A., Höhna, S., Larget, B.,
Liu, L., Suchard, M. A., and J. P. Huelsenbeck. 2012. MrBayes 3.2: Efficient Bayesian
phylogenetic inference and model choice across a large model space. Systematic Biology
61:539–542.
Royer, A., Malaizé, B., Lécuyer, C., Queffelec, A., Charlier, K., Caley, T., and A. Lenoble.
2017. A high-resolution temporal record of environmental changes in the Eastern
Caribbean (Guadeloupe) from 40 to 10 ka BP. Quaternary Science Reviews 155:198–
212.
Rozas, J., Ferrer-Mata, A., Sánchez-Del Barrio, J. C., Guirao-Rico, S., Librado, P., Ramos-
Onsins, S. E., and A. Sánchez-Gracia. 2017. DnaSP 6: DNA Sequence polymorphism
analysis of large datasets. Molecular Biology and Evolution 34:3299–3302.
106
Sánchez-Montes, G., Recuero, E., Barbosa, A. M., and Í. Martínez-Solano. 2019.
Complementing the Pleistocene biogeography of European amphibians: Testimony from
a southern Atlantic species. Journal of Biogeography 46:568–583.
Schwartz, A., and R. W. Henderson. 1991. Amphibians of the West Indies: Descriptions,
Distribution, and Natural History. University of Florida Press, Gainesville.
Silva, G. A. R., Antonelli, A., Lendel, A., Moraes, E. D. M., and M. H. Manfrin. 2017. The
impact of early Quaternary climate change on the diversification and population
dynamics of a South American cactus species. Journal of Biogeography 45:76–88.
Slatkin, M. 1987. Gene flow and the geographic structure of natural populations. Science
236:787–792.
Taberlet, P., Fumagalli, L., Wust-Saucy, A., and J. Cosson. 2002. Comparative phylogeography
and postglacial colonization routes in Europe. Molecular Ecology 7:453–464.
Tajima, F. 1989. Statistical method for testing the neutral mutation hypothesis by DNA
polymorphism. Genetics 123:585–595.
Thomas, R. 1999. The Puerto Rico Area, pp. 169–179. In: Crother, B. I. (ed.), Caribbean
Amphibians and Reptiles. Academic Press, San Diego.
Townsend, J. H., Eaton, J. M., Powell, R., Parmerlee, Jr., J. S., and R. W. Henderson. 2000.
Cuban Treefrogs (Osteopilus septentrionalis) in Anguilla, Lesser Antilles. Caribbean
Journal of Science 36:326–328.
Truong, H. T., Ramos, A. M., Yalcin, F., de Ruiter, M., van der Poel, H. J. A., Huvenaars, K. H.
J., Hogers, R. C. J., van Enckevort, L. J. G., Janssen, A., van Orsouw, N. J., and M. J. T.
van Eijk. 2012. Sequence-based genotyping for marker discovery and co-dominant
107
scoring in germplasm and populations. PLoS One 7:e37565
https://doi.org/10.1371/journal.pone.0037565.
Vasconcellos, M. M., Colli, G. R., Weber, J. N., Ortiz, E. M., Rodrigues, M. T., and D. C.
Cannatella. 2019. Isolation by instability: Historical climate change shapes population
structure and genomic divergence of treefrogs in the Neotropical Cerrado savanna.
Molecular Ecology 28:1748–1764.
Vences, M., Vieites, D. R., Glaw, R., Brinkmann, H., Kosuch, J., Veith, M., and A. Meyer. 2003.
Multiple overseas dispersal in amphibians. Proceedings of the Royal Society of London
B-Biological Sciences 270:2435–2442.
Wade, M. J., and D. E. McCauley. 1988. Extinction and recolonization: their effects on the
genetic differentiation of local populations. Evolution 42:995–1005.
Wang, S., Zhu, W., Gao, X., Li, X., Yan, S., Liu, X., Yang, J., Gao, Z., and Y. Li. 2014.
Population size and time since island isolation determine genetic diversity loss in insular
frog populations. Molecular Ecology 23:637–648.
Warken, S. F., Scholz, D., Spötl, C., Jochum, K. P., Pajón, J. M., Bahr, A., and A. Mangini.
2019. Caribbean hydroclimate and vegetation history across the last glacial period.
Quaternary Science Reviews 218:75–90.
Weigelt, P., Steinbauer, M. J., Cabral J. S., and H. Kreft. 2016. Late Quaternary climate change
shapes island biodiversity. Nature 532:99–102.
White, T. A., and J. B. Searle. 2007. Genetic diversity and population size: island populations of
the common shrew, Sorex araneus. Molecular Ecology 16:2005–2016.
108
Whitlock, M. C., and D. E. McCauley. 1990. Some population genetic consequences of colony
formation and extinction: Genetic correlations within founding groups. Evolution
44:1717–1724.
Wright, S. 1931. Evolution in Mendelian populations. Genetics 16:97–159.
Wright, S. 1943. Isolation by distance. Genetics 28:114–138.
Zeisset, I., and T. J. C. Beebee. 2008. Amphibian phylogeography: a model for understanding
historical aspects of species distributions. Heredity 101:109–119.
109
Curriculum Vitae
Andre Nguyen
Personal Information Email: [email protected]
Education SAN DIEGO STATE UNIVERSITY, San Diego, CA. Bachelor of Science in Biology, Emphasis in Zoology, Spring 2015 UNIVERSITY OF NEVADA LAS VEGAS, Las Vegas, NV. Master of Science in Biology (Ecology and Evolution): Fall 2017 - present (Research Advisor: Dr. Javier A. Rodríguez) Publications Nguyen A., Richart, C.H. & Burns K.J. (2015) Black-chested Mountain-Tanager
(Cnemathraupis eximia), Neotropical Birds Online (Schulenberg TS, Ed). Ithaca: Cornell Lab of Ornithology, Ithaca, New York, USA.
Professional Experience 11/2019 Assistant Instructor, Data Carpentry Genomics Workshop 8/2019 – present Discussion Section Instructor, Biology 415 - Evolution University of Nevada, Las Vegas 3/2019 – 5/2019 Field technician BEC Environmental INC., Las Vegas, NV. 9/2017 – 6/2019 Laboratory Course Instructor, Biology 197 - Principles of Modern Biology II University of Nevada, Las Vegas
9/2016 – 3/2017 Naturalist Arrowhead Ranch Outdoor Science School, Lake Arrowhead, CA.
Responsible for teaching 5th, 6th, and 7th grade science curriculum provided by the California State Board of Education
3/2016 – 5/2016 Avian Field Technician TW Biological Services, Oceanside/Camp Pendleton, CA.
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10/2015 – 11/2015 Research Associate Great Basin Institute, Death Valley, CA. 5/2015 – 08/2015 Intern (Research Assistant) Death Valley National Park, CA. Awards
Awards: Dean’s List – Fall 2014 & Spring 2015 (San Diego State University)
2018 Aquatic Biology Endowment ($5000)
2018 Graduate Professional Student Association Research Sponsorship ($689)
2018 Nevada INBRE Service Award ($5000)
2018 Nevada INBRE Service Award ($2300)