B I O L O G I C A L C O N S E RVAT I O N 1 4 2 ( 2 0 0 9 ) 4 1 4 –4 2 3

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The effects of forest fragmentation on euglossine bee

communities (Hymenoptera: Apidae: Euglossini)

Berry J. Brosi*
Department of Biology, Stanford University, 385 Serra Mall, Stanford, CA 94305, USA

A R T I C L E I N F O A B S T R A C T

Article history: Despite scientific and media attention on pollinator declines, there is still only a rudimen-
Received 22 July 2008 tary understanding of the response of bees—the most important group of pollinators
Received in revised form worldwide—to ongoing land use changes. Euglossine bees are an ecologically important
28 October 2008 Neotropical clade of forest-dependent pollinators. Despite the fact that euglossines are
Accepted 4 November 2008 well studied relative to other groups of tropical bees, only three previous studies, all from
Available online 17 December 2008 Brazil, address the response of euglossines to forest fragmentation. In this study, I tripled
the maximum sample size of previous efforts by sampling male euglossines in 22 forest
Keywords: fragments ranging in area from 0.25 ha to 230 ha in southern Costa Rica, using chemically
Forest fragmentation baited Van Someren traps. Abundance of euglossine bees was significantly positively
Costa Rica related to forest fragment size, negatively related to shape (edge:area ratio), and margin-
Orchid bees ally related to fragment isolation. Euglossine species richness showed similar, but weaker
Pollinators trends: richness was significantly positively related to the quantity of forest edge, margin-
Tropical wet forest ally negatively related to fragment area, and not related to fragment isolation. The positive
Van Someren trapping relationship between euglossine richness and abundance and forest fragment edge is
consistent with other studies that have found high euglossine density in secondary or
disturbed forest. The data suggest that individual euglossines move between forest frag-
ments, as has been shown in other systems. Still, forest fragmentation appears to affect
euglossine bees more strongly than other bee groups in the study region. Their large flight
range and positive relationship with forest edges may help to buffer the negative effects of
fragmentation, allowing euglossines to utilize even the very smallest forest fragments in
the study area.
Ó 2008 Elsevier Ltd. All rights reserved.

‘‘The dwindling and extinction of orchid bees, and their
plants, is one of the most striking traits of the decay of for-
est fragments melting on the Neotropical agroscape
today’’—Daniel Janzen (in Roubik and Hanson, 2004, p. 12)
1. Introduction
Given the importance of bees—the most important taxon of
pollinators—to both the persistence of native plant communi-
ties and the success of the human agricultural enterprise, it is
imperative that we better understand their responses to
ongoing global changes. This is particularly true in light of
the lack of scientific consensus on putative pollinator declines
(e.g. Cane, 2001; Ghazoul, 2005; Steffan-Dewenter et al., 2005).
Though land-use changes such as deforestation could hypo-
thetically have major impacts on bee communities (Buch-
mann and Nabhan, 1996; Allen-Wardell et al., 1998),
empirical studies of the effects of habitat fragmentation on
bees are scarce (Cane, 2001). Unfortunately, this paucity of

* Tel.: +1 650 450 3715; fax: +1 650 723 5920.

E-mail address: bbrosi@stanford.edu
0006-3207/$ - see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biocon.2008.11.003
 B I O L O G I C A L C O N S E RVAT I O N
studies is especially pronounced in the tropics, where nearly
all tropical-rainforest trees are animal-pollinated (Bawa, 1990)
and where pollen limitation can be particularly severe
(Vamosi et al., 2006).
The euglossine, or orchid, bees (Hymenoptera: Apidae:
Euglossini) are a particularly important group of Neotropical
pollinators. Euglossines pollinate many tropical plant groups,
including many economic plants (Dressler, 1968; Dressler,
1982), and are ecologically critical in their role in long-dis-
tance transport of pollen between widely spaced individuals
of low-density tropical plants (Janzen, 1971; Williams and
Dodson, 1972). Most euglossines spend the bulk of their time
in tropical forest habitats (Dressler, 1982), and as such are
thought to be particularly vulnerable to the effects of defores-
tation (Roubik and Hanson, 2004, p. 38). This is especially true
relative to other bee groups that extensively nest and forage
in human-altered habitats (Banaszak, 1992; Marlin and La-
Berge, 2001; Tylianakis et al., 2005; Cane et al., 2006; Brosi
et al. 2007, 2008). Still, a long-term effort in a region of Pan-
ama with relatively stable land cover found no declines in
euglossine bees over 21 years of regular sampling (Roubik,
2001). Euglossines are also well suited for forest fragmenta-
tion studies, because they are well described taxonomically
in relation to many other bee groups (particularly for the tro-
pics), and because euglossine males are attracted to chemical
baits and thus can be sampled thoroughly and easily (Dress-
ler, 1982; Ackerman, 1983).
Despite the importance of euglossine bees, only three pub-
lished studies have directly examined how they are affected
by forest fragmentation (Powell and Powell, 1987; Becker
et al., 1991; Tonhasca et al., 2002a,b). These studies were well
designed and executed, but wider inference from them is lim-
ited in several ways. First, all three took place in Brazil,
including two (Powell and Powell, 1987; Becker et al., 1991)
at the same site, the biological dynamics of forest fragments
(BDFF) project near Manaus (Bierregaard, 2001). The work of
Powell and Powell (1987) took place immediately after experi-
mental deforestation at the BDFF site, and the acute distur-
bance of forest clearing was likely the predominant factor
shaping their results (Becker et al., 1991; Cane, 2001). Second,
all three studies were based on a small number of sample
sites (Powell and Powell, four; Becker et al., seven; and Tonha-
sca et al., nine, three in forest fragments and six in continu-
ous forest). Third, while all three studies examined the
effects of forest fragment size, none of them assessed other
potentially important spatial factors, such as fragment shape,
isolation, or landscape context. Fourth, results from the stud-
ies, taken as a whole, are inconclusive. Becker et al. and
Tonhasca et al. found no significant relationships between
forest fragment area and euglossine diversity or abundance,
while Powell and Powell (whose results were likely dominated
by disturbance) did find a significant relationship. Finally, all
took place in landscapes dominated by large, contiguous
patches of forest. Since some euglossine bees possess flight
ranges of tens of kilometers (Janzen, 1971), these studies
may tell us little about the impacts of fragmentation on
euglossine bees in much more extensively deforested
landscapes.
In order to address these issues to improve understanding
of the responses of euglossine bees to forest fragmentation, I
1 4 2 ( 2 0 0 9 ) 4 1 4 –4 2 3 415
sampled male euglossine bees in 22 forest fragments ranging
over four orders of magnitude in area in a largely deforested
landscape in southern Costa Rica, thus tripling the sample
size of any previous study. I hypothesized that euglossines
would be more diverse and abundant in forest patches that
were: (1) larger in area; (2) less isolated from other forest; (3)
relatively low in edge:area ratio; and (4) in areas with more
surrounding forest cover.
2. Methods
2.1. Site selection
I sampled male euglossines in the landscape around the Las
Cruces Biological Station (8°47’N, 82°57’W) in the Valle de Coto
Brus, Puntarenas province, Costa Rica (Fig. 1). The 22 sampled
forest fragments ranged in area from 0.25 ha to 290 ha and
in altitude from 900 to 1400 m above sea level (Table 1). All
study fragments were situated in a matrix of primarily cattle
pasture (>60% of perimeter boundary). I chose sites separated
by at least 500 m (maximum site separation 13.5 km) and with
a minimum distance of 500 m to the nearest other forest frag-
ment of any size. The study fragments were all isolated by
deforestation approximately 40 years ago, when a wave of
European settlement in the 1960s reduced forest cover in
the region to the approximately 15% that it is today. Locally
collected pollen records, however, indicate several thousand
years of forest clearing, burning, and agriculture by indige-
nous people (Clement and Horn, 2001). The predominant land
uses in the region are cattle pastures, coffee fields, and small
agricultural plots for subsistence crops like corn, beans, and
bananas. See Brosi et al. (2008) for more details on the study
area and sampled fragments.
Euglossine bees are strongly forest-associated in this study
region. Extensive and taxonomically broader bee community
surveys in the same landscape (Brosi et al., 2007, 2008), sam-
pled no euglossines outside of forests in work that tallied
>4000 bee individuals. In thousands of hours of field time in
the study region, I have observed (non-baited) euglossine bees
foraging outside of tree-covered areas fewer than five times.
At least some species of euglossines, however, can be at-
tracted to chemical baits set out in deforested areas (B. Brosi,
pers. obs.; Milet-Pinheiro and Schlindwein, 2005).
While an ideal study design would sample each fragment
proportionally to its area, limited time and resources dictated
that I maximize the true sample size of the study (number of
forest fragments sampled) by sampling each fragment in one
place. The sole exception was the large (230 ha) forest frag-
ment at the Las Cruces Biological Station, which I sampled in
two sites 1 km apart.
2.2. Trapping
I sampled bees with traps, which allowed me to sample in
multiple sites simultaneously. I did not sample by hand-net-
ting at chemical baits, since in test trials a large proportion
of bees (commonly 20% or more) eluded either capture or
in-flight identification. Instead, I used Van Someren traps,
which are commonly used to sample butterflies (e.g. Horn-
er-Devine et al., 2003) and which consist of a cylinder of mesh
416 B I O L O G I C A L C O N S E RVAT I O N 1 4 2 ( 2 0 0 9 ) 4 1 4 –4 2 3

nicaragua

atlantic ocean

san josé

costa rica
pacific ocean

las cruces

panamá

Fig. 1 – Map of study site within Costa Rica.

Table 1 – Study sites. Sites 20 and 21 both represent the forest fragment at the Las Cruces Biological Station; all other
samples occur in unique fragments.

Site Area (Ha) Perimeter (m) Isolation (m) Elevation (m asl)

1 0.24 242.99 1203.4 1180

2 0.47 419.98 829.8 1089
3 0.53 354.54 778.0 963
4 0.65 409.06 832.9 1195
5 1.05 538.86 1838.3 1100
6 1.06 484.91 874.2 1027
7 1.13 543.01 1076.3 1115
8 1.18 509.40 1461.9 1160
9 1.46 711.29 507.0 1387
10 2.14 617.75 770.6 1284
11 3.27 1000.86 571.6 1184
12 3.39 879.19 850.2 898
13 4.67 1273.65 505.0 1000
14 6.83 1115.19 587.0 899
15 8.36 1706.42 569.7 1103
16 24.76 3730.86 512.0 1479
17 24.96 2791.96 519.0 1181
18 28.79 3532.11 769.4 916
19 55.19 5800.42 571.9 942
20 236.62 9737.84 599.5 1201
21 236.62 9737.84 548.4 1273
22 296.49 17,134.14 812.2 1414

cloth, raised 4 cm over a platform on which bait is placed. traps for sampling bees; Smith-Pardo (1999) used such traps,
Insects are attracted to the bait and fly upward, becoming baited with rotten fish, to sample bee communities (but not
trapped in the mesh cylinder, from when they can be col- specifically euglossines) in Colombian dry forest. I did not
lected alive. This is the second recorded use of Van Someren use McPhail traps (Bennett, 1972) because of their low catch
 B I O L O G I C A L C O N S E RVAT I O N
rates and the wetted, damaged bee specimens they yield,
which can be difficult to identify (Becker et al., 1991; Roubik,
2001).
For each sampling event, I set one Van Someren trap
15 m inside the forest fragment between 7:00 and 10:30am
and collected it approximately 3 h later, before daily rains
commenced. I rotated the temporal order of site sampling
within days, as well as the choice of essential oil bait, to min-
imize biases. I divided the sites into four ‘‘routes’’ of five or six
nearby sites and sampled each route on the same day, thus
running traps simultaneously in the sites nearest one
another.
I baited the Van Someren traps with either methyl salicy-
late (wintergreen oil, four drops) or cineole (eucalypus oil,
six drops) placed on paper towels in a small tupperware-style
plastic container on the trap base. I choose these two baits be-
cause in combination they attract nearly all recorded species
in the nearby (though lower-elevation) Osa Peninsula (Janzen,
1981). I purposefully utilized smaller quantities of essential oil
baits than have been used in some previous studies. In typical
euglossine sampling, a drink coaster or paper towel is com-
pletely saturated with essential oil, and bees are netted by
hand. In many contexts, this technique can lure hundreds
of bees in a very short time frame (e.g. Janzen, 1981). Because
destructive sampling was necessary to ensure accurate iden-
tifications, a larger bait quantity combined with the repeated
sampling protocol I used could have had significant deleteri-
ous impacts on local euglossine populations (Roubik, 1993).
Additionally, though the attractive radius of essential oil baits
has not been studied, a smaller bait quantity should hypo-
thetically have a smaller radius of attraction, thus allowing
for a more precise spatial focus on the targeted forest frag-
ments (rather than luring individuals from more distant,
non-target areas).
I sampled bees from June to September 2004, during the
wet season, the time of year in which Becker et al. (1991)
found the greatest richness and abundance of orchid bees,
and repeated trapping six times in each site. I collected all
bees within the traps and retained all specimens to maximize
the quality of identifications and to minimize bias from bees
returning to traps. I identified bee specimens using Roubik
and Hanson (2004); I. Hinojosa, University of Kansas, reviewed
and corrected species determinations and D. Roubik, Smith-
sonian Tropical Research Institute, reviewed the final species
list. Some specimens are deposited at the University of Kan-
sas Entomological Museum (SEMC); the remainder are held
at the Biology Department, Stanford University.
2.3. Measurement of forest characteristics
I measured area and perimeter by outlining forest fragments
from georeferenced, orthorectified aerial photographs (cour-
tesy G. Durán, Organization for Tropical Studies) in ArcMap
9.1 (ESRI, Redlands, CA, USA). For fragments <1 ha, I com-
pared these measurements to those made on the ground with
compass, laser range finder, and clinometer; the two methods
generated measurements within 5% of one another. As a
measure of forest fragment shape, I used the ratio of perime-
ter2:area, which is constant for circles of varying area. I de-
fined isolation as distance to the nearest forest fragment of
1 4 2 ( 2 0 0 9 ) 4 1 4 –4 2 3 417
any size, and calculated isolation distances using the Nearest
Features 3.8a (Jenness, 2004) extension for ArcView 3.2 (ESRI,
Redlands, CA, USA). I defined landscape context as the pro-
portion of forest from landsat data (classification details in
Daily et al., 2001) surrounding sample points at radii from
200 to 2000 m, which I quantified using ArcMap 9.1.
2.4. Data analysis
I compared forest characteristics (forest fragment area,
shape, isolation, and context) with bee relative abundance
(raw number of trapped bees) and diversity (measured by
both species richness and the Shannon–Weiner diversity in-
dex, hereafter SWDI). I used mixed-effects modeling to ana-
lyze data because of the hierarchical nature of the sampling
(repeated trapping sub-samples within sites). Such models
allow for the use of all data while correcting for pseudore-
plication below the site level (e.g. Crawley, 2002, 669). Forest
fragment area, shape index, and isolation distance were all
significantly correlated with one another, which would have
led to poorly-conditioned multivariate models. I therefore
tested these landscape factors in separate models and Bon-
ferroni-corrected the resulting P-values. The type of essen-
tial oil bait (cineole vs. methyl salicylate) was strongly
related to both euglossine diversity and abundance in indi-
vidual trapping trials, so I included bait type as an explan-
atory variable along with the individual landscape elements
in each of the analyses. Since the data are count-based and
sample variance increased with sample mean, I used gener-
alized linear models with Poisson errors in the mixed-effect
models (e.g. Olofsson and Shams, 2007), with site as a ran-
dom effect.
To assess the effect of landscape context on the euglossine
community, I calculated the correlation between the propor-
tion of forest surrounding sample points and euglossine spe-
cies richness and abundance, at scales from 200 to 2000 m
surrounding sample points (Ricketts et al., 2001; Steffan-Dew-
enter et al., 2002). To assess differences in community compo-
sition, I used Mantel and Partial Mantel tests to compare pair-
wise, standardized differences in landscape factors with Mor-
isita–Horn community similarity matrices. I used Moran’s I to
test for spatial autocorrelation in bee diversity and abun-
dance between sites.
To avoid pseudoreplication, I averaged data within sites for
all analyses except the mixed-effects models, in which I spec-
ified the hierarchical nature of the data without averaging. I
analyzed data using the R statistical programming language
(R Development Core Team, 2008), using the lme4 package
(Bates, 2008) for mixed-effects generalized linear models
and the vegan package (Oksanen et al., 2006) for community
ecological analyses.
3. Results
3.1. Overview
I sampled 412 euglossine bees in the genera Euglossa (18 spe-
cies) and Eulaema (5 species), Table 2. Two species, Euglossa
championi and Eulaema bombiformis, accounted for almost half
of the sampled bee individuals; Fig. 2 shows the rank abun-
418 B I O L O G I C A L C O N S E RVAT I O N 1 4 2 ( 2 0 0 9 ) 4 1 4 –4 2 3

Table 2 – Species list. Species in alphabetical order, sorted by genus. Proportion is proportion of total captures represented
by that species. See Roubik and Hanson (2004) for nomenclatural authorities.

Genus species Captures

Cineole Meth. Salic. Proportion (%) # Sites

Euglossa allosticta 1 0.2 1

Euglossa asarophora 1 0.2 1
Euglossa bursigera 1 0.2 1
Euglossa championi 12 107 26.1 18
Euglossa cognata 1 0.2 1
Euglossa crassipunctata 1 0.2 1
Euglossa cybelia 19 4.2 11
Euglossa despecta 1 0.2 1
Euglossa erythrochlora 83 18.2 16
Euglossa flammea 19 4.2 12
Euglossa gorgonensis 8 1.8 7
Euglossa heterosticta 2 2 0.9 3
Euglossa imperialis 1 2 0.7 3
Euglossa maculilabris 39 1 8.8 8
Euglossa mixta 25 5.5 14
Euglossa oleolucens 6 1.3 4
Euglossa sapphirina 3 0.7 3
Euglossa villosiventris 1 0.2 1
Eulaema bombiformis 106 23.2 19
Eulaema cingulata 1 0.2 1
Eulaema meriana 9 2.0 8
Eulaema nigrita 3 0.7 3
Eulaema speciosa 1 0.2 1
Additional species caught with hand netting at baits (not analyzed statistically)
Euglossa cyanura
Euglossa deceptrix
Euglossa dodsoni
Euglossa igniventris
Euglossa tridentata

140 cies in the system. The start time and duration of individual
trapping trials were not related statistically with euglossine
120
bee diversity or abundance.
100

80 3.2. Spatial and altitudinal autocorrelation

60

40 Bee abundance was significantly spatially autocorrelated

20
0 as measured by the Morisita–Horn index, was also signifi-
Fig. 2 – Rank abundance curve.
dance curve. Sub-samples were marked by high variability
within sites. For example, in one site a trap yielded four indi-
vidual bees of three species; 5 days later at the same site, the
trap yielded 38 bees of five species. Overall, there was a 5-fold
difference in species richness and a 15-fold difference in
abundance in the highest and lowest samples within a site.
Species accumulation curves showed a flattening trend
(Fig. 3), but also indicate that I likely did not detect every spe-
(Moran’s I = 0.10, P = 0.030), but bee diversity did not show
any pattern of spatial autocorrelation. Community similarity,
cantly spatially autocorrelated (Mantel r = 0.21; P = 0.013). I
found no relationships between euglossine bee diversity or
abundance and the elevation of sample sites. Community
similarity was, however, strongly related to elevation (Mantel
standardized r = 0.38; P < 0.001).
Because pair-wise site proximities and elevations were
also strongly correlated to one another (Mantel r = 0.43,
P < 0.001), I also assessed the partial correlations of elevation
and geographic distance on euglossine bee community simi-
larity. Holding geographic distance constant, community sim-
ilarity and elevation were still significantly correlated (Mantel
r = 0.33, P = 0.001). In contrast, holding elevation constant, bee
community similarity and geographic distance were no long-
er significantly correlated (Mantel r = 0.052, P = 0.28). There-
fore, the spatial autocorrelation in bee community similarity
appears to be governed by elevation, not geographic distance
 B I O L O G I C A L C O N S E RVAT I O N 1 4 2 ( 2 0 0 9 ) 4 1 4 –4 2 3 419

Number of species 25 Euglossine Species Accumulation 4. Discussion

20
15
10
5 a significant positive association with forest fragment edge, a
0
0 20 40 60 80
Number of samples
Fig. 3 – Species accumulation curve, based on randomizing
sample days 500 times without replacement.
per se. In subsequent analyses, I assumed that sites repre-
sented independent samples.
3.3. Landscape variables
Table 3 and Fig. 4 show the results for relationships between
forest characteristics and euglossine abundance and species
richness. Forest fragment area was significantly positively re-
lated to euglossine abundance (Fig. 4a), and marginally re-
lated to species richness (Fig. 4b). Shape (edge:area ratio) of
fragments was strongly related to both abundance (Fig. 4c)
and species richness (Fig. 4d) of euglossines, though in some-
what unexpected ways. Overall euglossine abundance in-
creased with increasing edge of forest fragments, as did
species richness. The direction of the response of species
richness, however, varied when samples were separated by
bait type. For cineole, richness maintained the same pattern
of increasing with fragment edge, but for methyl salycilate,
richness followed the previously expected pattern of decreas-
ing species richness with increasing edge. Fragment isolation
was marginally related to euglossine abundance (Fig. 4e), and
was unrelated to species richness (Fig. 4f). Euglossine abun-
dance was significantly positively related to proportion of for-
est surrounding sample sites at small scales, in radii around
sample points from 200 to 400 m (Fig. 5). At this scale, how-
ever, landscape context is essentially related only to the area
of the target fragment, since each study forest patch was sep-
arated from other forested areas by at least 500 m.
Euglossine bee communities are affected by forest fragmenta-
tion in southern Costa Rica. Euglossine abundance was signif-
icantly positively related to fragment area and forest edge,
and marginally related to fragment isolation. Euglossine spe-
cies richness showed similar but slightly weaker trends, with
marginal positive relationship to fragment area, and no rela-
100 120 140 tionship with fragment isolation. Euglossine community sim-
ilarity was related only to site elevation, when the correlation
between site proximity and elevation was considered.
This work was intended to examine the differences in
euglossine communities between different forest fragments
in a relatively small region, and not to estimate the true spe-
cies richness or diversity for the euglossine community as a
whole in the study region. Especially because of the novel
sampling method, the purposefully small bait quantities
used, and the incomplete species-accumulation curve from
the sampling (Fig. 3), the results of this work should not be
used to compare the species density of this region with other
areas. In addition, this work should be primarily considered in
the context of the common species that were sampled, as
many of the rare species were trapped in such low numbers
as to make interpretation difficult (Table 2).
The appropriate spatial pattern of sampling of euglossine
bees is the subject of some controversy, and could have
potentially impacted my results. Armbruster (1993) hypothe-
sized that euglossine males have relatively fixed discrete ter-
ritories over which they forage, and thus that sampling in
proximal sites within the same forest could yield substan-
tially different euglossines abundance and diversity. He sub-
stantiated this hypothesis with data from paired sample
sites 200 m apart in several Neotropical regions, which
showed statistically significant differences in the proportions
of euglossine taxa arriving at baits. This is of particular inter-
est here because in the interest of maximizing the true sam-
ple size, I sampled each forest fragment in one place, except
for the 230 ha patch at the Las Cruces Biological Station,
which I sampled in two sites.
The balance of published evidence, however, goes against
Armbruster’s ‘‘within-habitat heterogeneity’’ hypothesis. Ton-
hasca et al. (2002b) disagreed with Armbruster’s choice of sta-

Table 3 – Landscape factors and euglossine bee species richness and relative abundance. Because of strong correlations
between landscape factors, each was tested separately in a model including bait type. Thus, the ‘‘bait type’’ columns
report on the significance of bait type in the model for each landscape factor (rows), and the ‘‘interaction’’ column reports
on the bait type*landscape factor interaction. For all models, df = 20 (22 sample sites), and the Bonferroni-corrected
a = 0.017.

Factor Factor Bait type Interaction

Z P Z P Z P

Species richness Log area 1.73 0.083 5.86 <0.001*** 1.35 0.177
Shape 2.32 0.021 5.39 <0.001*** 2.48 0.013*
Isolation 1.05 0.295 1.57 0.117 0.99 0.322

Relative abundance Log area 3.13 0.002** 8.79 <0.001*** 1.75 0.080.
Shape 4.49 <0.001*** 9.52 <0.001*** 4.93 <0.001***
Isolation 1.74 0.082. 3.23 0.001** 0.81 0.417
420 B I O L O G I C A L C O N S E RVAT I O N 1 4 2 ( 2 0 0 9 ) 4 1 4 –4 2 3

abundance species richness

a Cineole Methyl Salicylate b Cineole Methyl Salicylate

species richness per subsample

abundance per subsample

6 6
30 30
fragment area

4 4
20 20

10 10 2 2

0 0 0 0

1 10 100 1 10 100 1 10 100 1 10 100

area (Hectares; log scale) area (Hectares; log scale)

c Cineole Methyl Salicylate d Cineole Methyl Salicylate

species richness per subsample

aabundance per subsample

6 6
30 30
fragment shape

4 4
20 20

10 2 2
10

0 0 0 0

20 40 60 80 100 20 40 60 80 100 20 40 60 80 100 20 40 60 80 100

shape (edge2:area ratio) 2
shape (edge :area ratio)

e Cineole Methyl Salicylate f Cineole Methyl Salicylate

species richness per subsample

6 6
abundance per subsample

30 30
fragment isolation

4 4
20 20

10 10 2 2

0 0 0 0

500 1000 1500 500 1000 1500 500 1000 1500 500 1000 1500

isolation (m) isolation (m)

Fig. 4 – Euglossine bee responses to landscape factors and bait type. While all subsamples are shown, these were grouped by
site for statistical analyses. Left column is abundance; right column is species richness. (a, b) Log10 area of forest (in hectares).
(c, d) Fragment shape (edge2: area ratio). (e, f) Fragment isolation (meters to nearest forest).

0.6
and euglossine richness or abundance)
abundance
0.5
(r-value; between % forest
0.4
correlation
0.3 richness
0.2
0.1
0
200 400 600
(circle radius in meters)
Fig. 5 – Correlation between landscape context (proportional
area of forest surrounding sample sites) and euglossine
species richness and abundance, at scales from 200 to
2000 m. Horizontal gray line shows cutoff for significant
correlations at P = 0.05 (r = 0.43); thus, only the correlation
between abundance and landscape context at radii from 200
to 400 m is significant.
tistical test, re-analyzed his data, and found no statistically
significant differences between the euglossine communities
in Armbruster’s within-habitat site pairs. Otero and Sandino
(2003) sampled four close-by sites in each of three distinct
habitats repeatedly over nearly two years in Colombia. While
they did not directly address the question of within-habitat
heterogeneity in their paper, Otero and Sandino’s data show
much larger differences between habitats relative to between
sample points within the same habitat. The within-habitat
samples appear from the tables and charts within their work
to have stabilized at relatively similar communities. Finally,
they marked and released each of the >2000 individuals they
caught, and yet recorded no recaptures.
My results and the literature are consistent with the idea
that individual euglossines routinely travel between forest
fragments in the area, rather than remaining resident in
individual forest fragments (or territories within fragments),
for several reasons. First, Tonhasca et al. (2003) recorded
marked euglossines (including some of the same species I
sampled) traveling between forest fragments in Brazil, and
it seems likely that euglossines would have similar behavior
in Central America. Second, I recorded euglossines even in
the very smallest fragments in the study region, which are
less than half of a hectare in area. Fragments of this size
are unlikely to provide all of the life-history requirements
needed for stable populations of several species of large,
fast-flying bees. Third, euglossines are known to have large
flight ranges, being capable of traveling up to 30 km in a
day (Janzen, 1971) and the fragments in this study area have
short isolation distances (less than 2 km in all cases).
Fourth, I did not find strong patterns of euglossine species
richness or abundance with isolation distance, which would
have been expected if euglossines were largely limited to
single fragments and only occasionally dispersed between
fragments. For example, research from true islands (isolated
B I O L O G I C A L C O N S E RVAT I O N
p = 0.05 significance line
800 1000 1200 1400 1600 1800 2000
spatial scale
1 4 2 ( 2 0 0 9 ) 4 1 4 –4 2 3 421
by water) has shown that where habitat isolation distances
are greater than typical flight ranges, isolation can have
strong effects on euglossine communities, particularly in
terms of species richness (e.g. Janzen, 1981). Fifth, it is unli-
kely that up to several hundred euglossines would arrive at
a single baiting station within a few hours (Janzen, 1981) if
they were more or less restricted to a small within-habitat
territory.
While it is thus likely that euglossines are moving be-
tween forest patches—a result that should hypothetically
weaken the relationships between forest characteristics
and the euglossine community—I still found strong relation-
ships between euglossines and both patch area and shape.
In terms of fragment area, given that nesting and foraging
resources are patchily distributed in tropical forests, more
bees should be found in larger fragments due to resource
availability alone. My results are consistent with this idea,
given that I found a significant positive relationship be-
tween forest fragment area and euglossine abundance, and
a marginal relationship with euglossine species richness.
The previous studies of euglossines and forest fragmenta-
tion were focused on fragment area (as opposed to other
landscape factors), and in general have suggested—while
not providing definitive evidence—that patch area impacts
euglossine bee communities. Neither Becker et al. (1991)
nor Tonhasca et al. (2002a,b) found significant differences
in euglossine bee abundance or diversity with forest frag-
ment area, though the smallest fragments in both works
had the lowest levels of euglossine abundance and diversity.
In both studies, the small number of fragments may have
limited their power to find statistical differences. In con-
trast, Powell and Powell (1987) did find significant relation-
ships between fragment area and euglossine richness and
abundance, but they sampled immediately after a major dis-
turbance (experimental deforestation), which is thought to
be the primary factor affecting their results (Becker et al.,
1991; Cane, 2001).
Of the landscape factors I tested, forest fragment shape
had the strongest associations with both euglossine species
richness and abundance. Euglossines are significantly more
abundant and speciose in forest fragments with more edge
relative to their area. This relationship also had a significant
interaction term with bait type for euglossine species rich-
ness. For bees attracted by cineole, the relationship was in
the same direction as the overall pattern (richness increased
with increasing edge), but for bees attracted by methyl salic-
ylate, the direction of the response changed, and species
richness decreased with increasing edge. This difference in
the direction of the response of species richness to edge
was surprising, because abundance and richness are typi-
cally positively related. This pattern could be explained in
part by the fact that the three dominant euglossine species
(which together comprised 2/3 of all sampled bees) were
all attracted primarily to methyl salicylate, and also were
all positively related to increasing forest edge. If in general
forest edges provide greater resource density, the dominant
species could potentially be competitively excluding less-
common species from the ‘‘edgier’’ fragments. Alternatively,
less-common euglossine species attracted by methyl salicy-
late could prefer forest-interior habitats, which are in scar-
422 B I O L O G I C A L C O N S E RVAT I O N
cer supply in ‘‘edgier’’ forest fragments. Why such a pattern
would not be shown for the other attractant (cineole), how-
ever, is puzzling.
Evidence does exist that euglossines will readily utilize re-
sources from less-than-pristine forest. Otero and Sandino
(2003) sampled three habitat types—farm, secondary forest,
and old logged forest—in coastal Colombia and found euglos-
sine abundance was positively related to land-use intensity.
The farm sites had the highest euglossine abundance, fol-
lowed by secondary forest, while the low-disturbance old
logged forest had the lowest euglossine abundance. Similarly,
Rincon et al. (1999) found that logged forest had higher
euglossine abundance than did less-disturbed forest in Costa
Rica. Both of these results are consistent with the idea that
forest edges may be sites of relatively high resource-density
for euglossine bees.
While euglossines are positively associated with forest
edge, they still appear to be more affected by forest frag-
mentation than other clades of bees in the region. Sampling
across all groups of bees in the same region, Brosi et al.
found that bee abundance and diversity were not related
to distance from forest (2007) or forest fragment size
(2008). Species composition, however, did change with both
distance to forest and forest fragment size. Net sampling
results from both within forest fragments and in paired
pasture habitats directly adjacent to them showed that
15% of sampled bees within forest were euglossines, com-
pared to 0% outside of forest (Brosi et al., 2008). Given this
and the positive significant relationship between forest frag-
ment size and euglossine abundance, and the marginal
relationship with species richness, euglossines are appar-
ently more affected by forest fragmentation than other
groups of bees.
A feature of both the work presented here and of previous
studies (e.g. Roubik, 2001) is high sample variability. My sam-
ples, taken typically within four to 6 days of one another in
the same site and repeated six times, show a similar magni-
tude of variation as studies conducted over several years (Rou-
bik, 2001) and indicate that this variability is not necessarily
due to fluctuations in population size. Euglossines may move
large distances in the landscape over short time scales in re-
sponse to floral or other resource availability, and/or may be
affected by relatively subtle changes in weather that are re-
flected in their presence at traps. Thus, while long-term mon-
itoring is indisputably valuable, perhaps the most important
factor for characterizing a euglossine bee community is re-
peated sampling of the same sites, even if those samples are
repeated over a relatively short space of time.
Understanding euglossine responses to habitat fragmen-
tation has been hampered by a small number of studies,
and future work should consider and potentially even focus
on the larger context of study sites. For example, research
in landscapes that are more extensively deforested may re-
veal stronger negative responses of euglossine species rich-
ness than previous work. Euglossine bees are a critically
important group of tropical forest-dependent pollinators,
and given the increasing concern over pollinator declines
worldwide, we must better understand how ongoing land-
use changes affect their abundance, diversity, and commu-
nity composition.
1 4 2 ( 2 0 0 9 ) 4 1 4 –4 2 3
Acknowledgements
I thank the many Costa Rican families who allowed me to
work on their land. G. Daily contributed crucial support, ad-
vice, and funding at all stages of this work. G. Daily, P. Ehrlich,
K. Levy, and R. Pringle gave helpful suggestions on earlier
drafts of the manuscript. I. Hinojosa, University of Kansas,
introduced me to orchid bee biology and identification,
helped inspire this study, and confirmed my species determi-
nations. D. Roubik reviewed and corrected the final species
list. J. DeNoyer, K. Frangioso, B. Graham, J. Ilama, F. Oviedo,
and T. Shih provided superb field assistance. The staff of the
Las Cruces Biological Station and the Organization for Tropi-
cal Studies (especially R. Quirós, E. Ramirez, and Z. Zahawi)
provided cheerful field research support. G. Durán of Las Cru-
ces provided key spatial data. J. Ranganathan provided GIS ad-
vice, data, and assistance. I am grateful for funding from the
Anne M. and Robert T. Bass Stanford Graduate Fellowship in
Science and Engineering, a Teresa Heinz Scholarship for Envi-
ronmental Research, and the Moore Family Foundation; the
Stanford University Field Studies and Human Biology Re-
search Experiences for Undergraduates (HB-REX) Programs;
and Grants to the Center for Conservation Biology at Stanford
University from the Koret, McDonnell, Sherwood, and Win-
slow Foundations and Peter and Helen Bing.
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