TROPICAL ECOLOGY, ASSESSMENT, AND MONITORING

(TEAM) INITIATIVE

ANT MONITORING PROTOCOL

Author
Puja Batra, Ph.D., Project Director
 Introduction
Ants constitute a staggeringly high proportion of the earth’s biomass (Fittkau and Klinge 1973) and species
diversity (Erwin 1989; Holldobler and Wilson 1990). They provide a number of crucial services important
to overall functioning of ecosystems, such as soil enrichment and turning, seed dispersal, control of
herbivorous insects, symbiotic interactions (Holldobler and Wilson 1990 and references therein), and have
been the subject of a vast amount of research over several decades which has increased our understanding
of ecology, evolutionary biology, animal behavior, and the list goes on. Thus, a multi-taxon monitoring
program that aims to capture change in elements known to be critical to overall biodiversity and ecosystem
health must necessarily include ants.

In addition to the potential value of ants as a surrogate group for overall diversity (see (Alonso 2000) for a
detailed discussion), they have great promise as indicators of ecological change (Nepstad 1995). Ant
community structure has been shown to be sensitive to heterogeneity in a number of environmental factors.
For example, data from both within and across site studies have shown relationships between ground
dwelling ant communities and soil properties (Catangui et al. 1996; Bandeira and Harada 1998;
Bestelmeyer and Wiens 2001), seasonality and climate factors (Bestelmeyer and Wiens 1996; Feener and
Schupp 1998; Kaspari and Weiser 2000), vegetation density (Bestelmeyer and Wiens 2001), vegetation
communities (Majer and Delabie 1994; Majer et al. 1997; Morrison 1998; Bestelmeyer and Wiens 2001),
and plant productivity (Kaspari et al. 2000; Kaspari et al. 2000). Many of these factors have been shown
to be currently undergoing measurable changes due to climate and land use change (Root and Schneider
2002; Walther et al. 2002; Parmesan and Yohe 2003), and these effects are predicted to continue into the
foreseeable future (Bazzaz 1998; Enquist 2002).

Many aspects of these factors will be monitored by the TEAM Initiative. Thus, the correlation of multiple
data sets taken in the same areas over the same time span will allow not only intrasite correlations between
ant communities and various biological, climatic, and physio-chemicalfactors, but also will allow for
detection of intersite trends that may be occuring regionally. Furthermore, by monitoring communities of
ants over time, we will add enormously to our understanding of the baseline levels of fluctuations in ant
diversity and turnover at different spatial scales.

Specific questions
By monitoring leaf litter ants several times per year, we will address the following questions:

1) What is the baseline level of fluctuation in leaf litter ant communities in tropical forests? How
does that baseline differ across regions?
2) Are there trends in indices of species richness or relative abundance over time? If so, do those
trends correlate with trends in climate measures, soil properties, tree growth rates,
phenology/litterfall, or measures of land-use change? At what spatial scale do trends occur over?
Are similar trends occurring across regions?
3) Is community composition changing over time? If so, in what way? (e.g., Are species being lost
or gained? Are there invasions of exotics occurring? Is community structure, i.e., dominance and
sub-dominance shifting? Are species which are relatively restricted in geographic range more
vulnerable to population changes?)
4) At what spatial scale does turnover in litter ant community composition occur?

Methods for monitoring ants

Because of their ubiquity and high species richness, there are many techniques and microhabitats useful for
ant collection. These include sampling from the leaf litter and/or soil using a variety of extractor types,
using pitfall traps to collect ants from the soil surface, setting baits to attract various foraging guilds,
canopy fogging, beating vegetation to collect ants in the shrub layer, hand searching, and still others. The
utility of these various techniques depends entirely on the questions being addressed, and results

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quantifying aspects of diversity may or may not be comparable across sites due to differences in collector
experience, uneven presence of certain guilds, etc. For the purposes of TEAM monitoring, the technique
chosen must provide a method which minimizes the effects of varying collector experience, maximizes
efficiency in species: effort ratio while also reliably capturing ants within the same species guild.

In a massive study conducted in the Atlantic forest region of Brazil which compared the efficiency in
capturing species richness of 17 different ant collection methods, Delabie, et al ( 2000) concluded that the
combination of pitfall traps and leaf litter sampling using Winkler extractors captured 50% of the total
species richness found. A standard protocol was agreed upon by many members of the myrmecologist
community (Agosti et al. 2000), and this protocol, known as the Ants of the Leaf Litter (ALL) method is
rapidly becoming the preferred method used around the world for inventory and monitoring of ants.
TEAM monitoring will utilize the ALL method, with some minor modifications in transect length and
spatial design. The modifications will not alter the comparability of TEAM data to other studies examining
species richness and abundance, since such estimates are typically made on a per sample basis, and not a
per transect basis. TEAM monitoring using ALL will be conducted at a massive scale, sampling four
transects of 100m length in each Integrated Monitoring Array (IMA) four times per year.

Spatial layout
The ant transects will run along side soil termite transects, thus this layout was designed to accommodate
both types. The sampling design for leaf litter ants consists of replicated 100 meter transects, with
sampling stations spaced at ten meter intervals along the transect. Each sampling season, four 100 m
transects will be placed inside the 1km2 Integrated Monitoring Array (IMA) (figure 1). For the purposes of
placement of ant (and termite) transects, the IMA can be considered to be divided into four quadrants of
equal area, with invisible lines running down the central horizontal and vertical axes. One transect will be
randomly placed within each of these quadrants. Constraining the placement of each transect’s potential
location to only one fourth of the IMA will allow for some degree of randomization while also achieving
even coverage of the array over time, thus maximizing the chance of capturing some of the heterogeneity of
microenvironments that the large IMA contains.

Transects will be placed in the grid square between the pair of (x,y) coordinates chosen and the next higher
number of each. For example, if the (x,y) coordinates chosen randomly are (04, 600), the transect will be
placed in the grid square that falls between points (04, 600) and (05, 700). The divisions between
quadrants run along line 06 and line 500 (figure 1.) Using a random number table, four sets of randomly
chosen (x,y) coordinates must be selected, with the coordinates falling between the following numbers:
01-05, 000-400
01-05, 500-900
06-10, 000-400
06-10, 500-900
Once a grid square is used, it will never be re-used until all possible locations have been used. Thus, after
the first season of sampling, it will be necessary to carry a list in the field of all the locations which have
been previously used and are therefore, not repeatable. If the grid square selected has previously been
sampled, random number selection must be repeated until an appropriate square is found.

If there is standing water in the middle of the grid square, shift anywhere within the same square to a dry
area so that the 100 meter transect can be placed perpendicular to the trail. If the amount or distribution of
standing water excludes the possibility of placingthe transect anywhere in the grid square, repeat the
random number selection process until a dry grid square is found. If all available choices (i.e., all non-
repeatable locations) have standing water in them, make a note of this in your notebook, and select a dry
repeat location. Similarly, if you reach the grid square and find that it is being swarmed by army ants,
make a note of it and repeat the selection process. The reason for this is not simply to avoid sampling an
overabundance of army ants, but more to facilitate the field work in the leaf litter without the obvious
problems to collectors that army ants might cause.

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The transect should run such that it starts along a major trail, i.e., perpendicular to the trail, halfway (50 m)
between the two corners. The first sampling station should be inside the grid square, 10 meters from the
trail, so that the last sampling station falls at the edge of the grid square where there may or may not be a
temporary maintenance trail running between IMA numbering stakes.

To avoid confusion, flagging for different groups of TEAM variables is color coded. All insect sampling
locations and/or entrance points from the trail should be marked with blue flagging only.

Equipment
(see Appendix 1 for a list of equipment suppliers)

For litter sampling (per transect):

random number table
sighting compass
100 m nylon rope knotted at 10 m intervals
blue flagging
1 1m2 quadrat made of wood or PVC (movable joints; one corner should be openable)
1 machete
2 pair gloves
1 small hand rake
1 small hand trowel
1 watch with seconds or stopwatch
1 litter sifter
10 litter stuff sacks (included with the mini-Winkler apparatus) and some extras
2 waterproof notebooks
pencils
large plastic bag, large enough to carry 10 mini-Winkler samples without compressing them

For pitfall traps (per transect):

soil auger (or sharp edged pipe with diameter of the pitfall cups) to dig pitfall trap holes; must be strong
enough to cut through roots.
10 durable plastic cups, 7-8 cm diameter, 8-10 cm depth, with no ridges inside or at mouth
1 liter 75% Ethanol
5 ml liquid detergent
10 small plastic plates, few cm larger than diameter of cups; used as a roof for the pitfall trap
thin wooden skewers
labels (card stock)
squeeze bottle
forceps
waterproof and Ethanol proof ink pen

For measuring ecological conditions

Kestrel 3000 Pocket Weather Station
Electronic soil probes for pH, temperature, humidity
Small ruler (15 cm)

For processing mini-Winkler litter samples and pitfall trap contents (per transect):
2 pieces nylon rope (5 m)
1 2x3 m plastic sheet (only 2 are needed in total)
10 mini-Winkler apparatus (each mini-Winkler contains a mesh bag, and the external cloth portion) and a
few extra
2 light colored plastic trays
medium width (5 cm) paintbrush

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fine (2-3 mm) paintbrush
forceps
10 plastic cups with no ridges inside or at mouth
2 liters 95% Ethanol (source alcohol, not denatured EtOH)
squirt bottle
10 vials (25-50 ml) with tight caps
1 large ziploc bag
labels (card stock)
waterproof and EtOH proof ink pen
scissors

Methods
Field sampling
Running the transect
Use a compass and a 100m nylon rope that has already been measured and marked with knots at 10 m
intervals. The first sampling station will be 10 m from the trail so tie the rope to a tree just inside the grid
off the trail. With the rope to help you run a straight line, sight intermediate landmarks through the sighting
compass that fall in the desired direction. The heterogeneity of microhabitats is important in maintaining
species diversity, so do not avoid any types of microhabitats that fall in the straight line, even though some
may be difficult to sample in. Place a piece of blue flagging at every 10 m increment until you have placed
10 such flags, all in a straight line. It is best to leave the rope in place, tied to trees at either end so that you
can easily find the next point without the need to search for the flagging. If there are trees, fallen logs, etc
in the line, the transect must include them. The knots (and flagging) will be the points at which you will
sample leaf litter and place pitfall traps.

Leaf litter samples

(modified, but largely after Bestelmeyer et al. (2000))

The leaf litter sampling is to be done when the litter is somewhat damp, but not saturated. Wait a few hours
after a heavy rain to collect litter. Transects are best done with two people working on the litter samples
and one person simultaneously placing the pitfall traps. Hereafter, the terms “Winkler” and “mini-
Winkler” are interchangeable.

Try as much as possible to walk on the side of the rope that will not be sampled, to avoid trampling or
disturbing the litter. At each flagged location on the transect, place the 1 m2 quadrat on the ground. To
ensure that your bias of having seen the entire transect while running the line does not influence where you
place the quadrat, decide in advance of running the transect line where relative to the 10m marks you will
place the 1m2 litter sampling quadrat. For example, decide on the rule that you will always place the
quadrat on the right side of the line, with the knot on the rope falling in the bottom corner of the quadrat. A
rule against this type of bias will result in being able to sample many of the microhabitats that you may
encounter on the transect such as treefall gaps, fallen logs, etc. The quadrat has one corner left open so that
it can be put around trees and shrubs that will fall in the sampling spot. If there are trees too large to put the
quadrat around, shift the quadrat so that one edge is touching the tree trunk. If there is a fallen tree on the
ground, include it if possible since it will probably have an accumulation of litter on it. If it is too large,
place the quadrat as close to it as possible so that one edge is touching the fallen log.

It is highly advisable to wear gloves while handling the leaf litter. Remove large sticks in the quadrat. Use
a machete to chop open decaying logs inside the quadrat. Scoop litter inside the quadrat toward the center,
using the hand trowel and hand rake (figure 2). Break open twigs and clods of soil. Tie off the bottom of
sifter, and place litter in top of sifter up to ¾ full. Break up any more twigs or pieces of rotting wood that
are in the sifter. Hold both handles and shake the sifter in all directions continuously for 30 seconds (figure

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3). This shakes the smaller litter and invertebrates in it to the bottom part. Thoroughly turn the litter over
by hand and shake for another 30 seconds.

Mark the cloth stuff sack with a label with the following information, written in waterproof and alcohol
proof ink:
Date: mm/dd/yr
Array: 1-6
Transect grid square coordinates: (x,y)
Sample number (1-10)
Winkler
Place the litter stuff sack below the sifter sack, and untie the sifter sack. Shake all contents of sifter into the
litter stuff sack (figure 3), shaking the cloth to dislodge any insects crawling up the sides of the sifter. Close
the sample bag tightly.

When carrying the litter samples back to the field station in a large plastic bag, take care not to compress
them or place anything on top of them. After returning to the field station, the samples will be placed into
mini-Winkler sacks and left for 48 hours during which time the ants will migrate out of the leaf litter into a
collection cup or bag. This process is described in a later section.

Pitfall traps
Pitfall traps will be placed along the same transect and at the same time as the litter sample collection, and
left in place for 48 hours. Place pitfall traps in a consistent place relative to the quadrat, about 10-15 cm in
front of the litter quadrat, in approximately the midpoint of the front edge of the quadrat. Using a soil auger
or metal pipe whose diameter is roughly the same as the plastic cups, dig a hole that is the same depth as
the cup. You will have to cut through roots in many instances. If the roots are impossible to cut through,
shift the location of the hole by a few centimeters. If possible, cut the hole without removing the litter.
This will make it easy to use the edge of the hole to measure litter depth (described below).

Place a plastic cup in the hole, flush with the ground (figure 4). If the hole is too wide, you will need to
pack in soil around the sides to keep it in place. In order to keep soil out of the cup (which gets into the
sample and is difficult to remove during sample processing), stack a second cup inside the one that will be
the trap while you arrange the soil around it. Also rearrange the leaf litter so that it comes up to the edge of
the pitfall trap.

On a label, using waterproof and alcohol proof ink, record the following information:
Date: dd/mm/yr
Array number: 1-6
Transect grid square coordinates: (x,y)
Sample number: 1-10
Pitfall
Place the label in the cup and pour in about 2-3 cm depth of 75% ethanol that has been mixed with a few
drops of liquid dish detergent (about 5 ml detergent per liter). The detergent breaks the surface tension so
that insects which fall into the trap actually sink into the liquid, and also prevents rapid evaporation of the
ethanol during the 48 hours that the trap is left in the field.

Cover the cup with a “roof” that is held a few centimeters above the cup. This can be made of easily
available materials, such as a small plastic plate held up with thin wooden skewers (figure 4). The roof
should be only slightly larger in diameter than the cup.

Leave traps in place for 48 hours. Collect them on day 3 by transferring the contents of the trap to a 50 ml
tube with the aid of a squeeze bottle of 75% ethanol and forceps, putting a label inside the tube, and closing
the tube tightly. Any labeling information that you write on the outside of the tube is likely to be erased if
the alcohol drips or spills on it, so be sure there is a clearly written label inside the tube, preferably
positioned so that it can be read from the outside of the tube.

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Ecological data
While collecting litter samples and placing pitfall traps, it is crucial to collect some basic data on ecological
conditions on the transect. Appendix 2 contains a copy of the data sheet that must be filled in completely
when a new transect is sampled.

When starting the sampling, using the Kestrel 3000 Pocket Weather Station, record the air temperature and
relative humidity. In three 10 m x 1m sections along the transect, collect data on the vegetation, canopy,
ground habitat, and leaf litter. The ten-meter sections where you will collect these data are in from meters
1-10, meters 40-50, and meters 90-100 along the transect line in a 1m wide band.

The vegetation description is a quantification of the number of live stems in each of the 10m2 area sections.
Count and record the number of live stems in three different height categories and one girth category:
< 1m height
< 2 m height
>2 m height
> 10cm DBH
Within any given section, if trees land in partially inside and partially outside the 10m2 area, alternate
whether you count it or not, i.e., count the first such tree in your description, exclude the second such tree,
and do on.

For canopy description, estimate the height in meters of the main canopy layer and the emergent layer.
Qualitatively describe the canopy cover as one of the following categories:
Closed
Partially open
Open with sub-canopy secondary growth
Open with very little or no sub-canopy growth (new gap)

For ground habitat description, estimate in increments of 10% the amount of ground cover of each of the
following categories:
Bare soil
Stone
Live vegetation, including roots
Decomposing wood
Leaf litter
In this case, ground cover refers literally to what is touching the ground. For example, if there is a treefall
whose trunk is not touching the ground, and under the trunk there is bare soil, the relevant category would
be bare soil. In cases where a category is represented by less than 10% but more than zero, record 1% as a
way to indicate its presence in very low relative amounts.

Leaf litter conditions also need to be recorded, as they are important in determining the immediate
environment that the ants experience. Using electronic soil probes, record the following:
Soil pH
Soil temperature
Soil humidity
Also measure the litter depth in cm with a small ruler. The easiest way to do this is from the hole that you
have dug for the pitfall trap, if you are able to cut the hole without disturbing the litter around it.

Extraction of ants from litter samples using Mini-Winkler apparatus

The following can be done in the field under a large tarp in a place that is protected from the wind and rain.
However, it is much easier if you bring the samples back to the field station. This process should be done
on the same day as the samples were collected in the field, and will be left in place for 48 hours. It is very
useful to make a note of which species of ants may be foraging in the room where you are working. They
are often invasives or extremely common ants that can be easily identified (W. Overal, pers.comm.) At
some later date, this information may be useful in discerning a contamination problem. If you are working
with the samples in a field shelter and not an enclosed building, you will need to be extremely vigilant
about making sure that ants do not crawl on the ground sheet on which you are working. They will be

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easily confused with escapees from your sample otherwise, and you will contaminate the samples with ants
that did not come from your transect.

Hang ten mini-Winklers from a rope strung horizontally in an area of a room where they will not be moved,
blown by the wind, or disturbed (figure 6). Tie an empty cup in the bottom of each. It will later be filled
with ethanol. In most instances, one mesh sack will be sufficient to hold a single litter sample. However,
in the event that the litter from one sample does not fit entirely in one mesh sack, it will be useful to have a
few extra mini-Winklers on hand.

Place a large plastic sheet on the ground. From now on, do all work on it so that escaping ants can be seen
and captured. Working above a plastic tray to catch litter that falls through the mesh, transfer the litter
sample from a single litter stuff sack to a mesh inlet sack. Use the 5 cm width paintbrush to collect the
litter that has fallen in the tray and transfer it back into the mesh sack (figure 5). You will have to repeat
this process several times to get all of the fine litter particles in. If ants escape during this process, pick
them up with either forceps or a fine paintbrush moistened with alcohol or water and put them into a vial or
back into the litter. Periodically shake the mesh sack to let the contents settle and to remove air pockets. Do
not leave any material in the tray or on the ground sheet. The mesh sack should remain flat even with the
litter in it in order for it to be effective, so do not stuff it so full that it bulges. Between samples, be sure to
clean the brushes and trays of any remaining debris particles, so that tiny ants which may be lodged in them
do not end up in the wrong sample.

Carefully hang the mesh sack inside the external mini-Winkler sack (figure 6). The mesh sack should not
touch the walls of the mini-Winkler sack. If any debris falls into the cup, empty the cup back into the mesh
sack. Partially fill the cup with 95% ethanol and tie this to the bottom of the mini-Winkler. Put the sample
label in the cup, and also transfer into the cup any ants that you caught while transferring the litter. The
ants will fall out of the mesh sack into the alcohol filled cup.

After 24 hours, each mesh sack must be carefully taken out, emptied into a plastic tray and put back into
the mesh sack again, following the same procedure outlined above. This process stirs up the litter and
allows more ants to be collected.

Leave the mini-Winkler apparatus in place for another 24 hours. At the end of the period, remove the cup,
pour the contents of it into a 50 ml plastic tube, rinsing cup with EtOH. Be sure to include label with the
sample. Close the tube tightly. Keep all tubes from a transect together in a large ziploc bag, also labeled.

Personnel needed for field work and sampling rotation

The entire process of running the transect line collecting and sifting 10 litter samples, placing 10 pitfall
traps, and taking ecological data takes a team of four people about two hours, after the initial process has
been learned. Two transects can be done in a single day.

Transferring 10 litter samples to mini-Winkler extractors takes two people about two hours also. Thus, a
typical day in the field and lab will take a team of four people 8-10 hours, including travel time to the
arrays, grid coordinate selection, and walking to the grid locations.

Since transects need to be revisited to pick up the pitfall traps 48 hours after the transect is run, the most
efficient way to arrange sampling of different arrays is to do it in such a way as to minimize the number of
trips to a single array. The following chart illustrates an efficient sampling rotation.

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IMA # transect # Day 1 Day 2 Day 3 Day 4 Day 5 (etc)
1 1 Transect Pick up pitfalls
1 2 Transect Pick up pitfalls
2 1 Transect Pick up pitfalls
2 2 Transect Pick up pitfalls
1 3 Transect Pick up pitfalls
1 4 Transect Pick up pitfalls
2 3 Transect
2 4 Transect
3 1 Transect
3 (etc) 2 Transect

Here, on days 1, 2, 3, and 4 only one array is visited. On day 5, two arrays are visited, and so on. In this
rotation, starting to sample transects in IMA 2 on day 2, before all transects on IMA 1 have been done
prevents an extra visit to IMA 1 since you can pick up pitfalls from transects 1 and 2 on IMA 1 at the same
time that you sample transects 3 and 4.

Sampling will be done four times a year: early rainy season, peak rainy season, early dry season, and peak
dry season.

Specimen processing and identification

Samples should be identified by an expert taxonomist, or under the supervision of one. Details regarding
this process will be specified in later versions of this document.

Field data forms, database forms, and data entry

A sample data sheet for collection of ecological data on the transects is included in Appendix 2. All data
should be entered into the TEAM database as soon as possible after collection. The fields in the database
are often constrained to a pulldown menu of only a given set of available choices. For example, The
following fields will be included in the database, along with the possible choices and their definitions. The
types of data fields that appear on the forms is of four types: transect information, ecological information,
specimen information, and species information. The data forms in the database are divided somewhat
differently, but the fields that appear are listed.

Information about the transect and its location

FIELD CHOICES DEFINITION
Array ID 1-6 Identity of the array

Transect Coordinates Identifies the grid square where

trap is located. One set of
coordinates identifies the square
because the second set of
coordinates is assumed to be the
next higher number of each.
Sampling type Winkler Type of sampling
Pitfall
Section of transect 1-10 Section of transect where
40-50 ecological data was taken
90-100

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Information about ecological conditions at time of sampling. (This will also include all the data listed on
field data sheet in Appendix 2.)
FIELD CHOICES DEFINITION
Date dd/mm/yr Date of sampling
Time of day hh:mm Time at start of sampling
Names of collectors Name1 Names of field workers
Name 2
Air temperature XX.X° C Air temperature at onset of
transect sampling
Relative humidity XX.X % Relative humidity at onset of
transect sampling
# stems < 2 m height Count data Number of stems counted in this
(etc for ecological data) category

Information about specimens sampled

Genus and species (list of possible species, including Identity of each specimen
morphospecies A,B, etc) collected. If a species is
encountered that does not appear
on the list, you must first enter the
species’ information in the
species table below. It will then
automatically appear in the
pulldown list.
Sex M Sex of specimen, if known
F
undetermined
Date determined dd/mm/yr Date of the identification of the
specimen
Determined by Name 1 Name of person who identified
etc the specimen

Information about species

FIELD CHOICES DEFINITION
Order Hymenoptera Only one order sampled

Family Formicidae Formicidae only is the target

group.

Sub-family list Sub-family

Genus and species Each species, even

morphospecies, needs a new entry
before being able to record its
specimens
Species authority Name of the author who first
described the species

There are many more fields for species specific information that is necessary for taxonomists and museum
collections, but for the purposes of this protocol, what is listed above is sufficient.

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 Data analysis
See Avian Protocol for a discussion of analytical methods of species richness estimators. Further analyses
will include ordination methods of diversity indices along axes that describe climate, soil, and vegetation
measures; examination of differences in community composition between years, and between regions using
rarefaction methods (Gotelli and Graves 1996), similarity indices, and ordination (Kremen 1992; Kremen
1994). More in-depth explanations of data analyses will be given in future versions of this document.

Acknowledgements
Discussions with Leeanne Alonso, Roberto Brandão, Brian Fisher, Ana Harada, and Jack Longino have
contributed to the ideas and methods described. Field tests with Ana Harada and Bill Overal helped to fine
tune many of important details of the protocol.

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 Literature Cited
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Monitoring Biodiversity, Smithsonian Institution Press.
Alonso, L. (2000). Ants as Indicators of Diversity. Ants: Standard Methods for
Measuring and Monitoring Biodiversity. D. Agosti, J. D. Majer, L. E. Alonso and
T. R. Schultz. Washington and London, Smithsonian Institution Press.
Bandeira, A. and A. Y. Harada (1998). "Densidade e distribuição vertical de
macroinvertebrados em solos argilosos e arenosos na Amazônia central." Acta
Amazonica 28(2): 191-204.
Bazzaz, F. A. (1998). "Tropical forests in a future climate: Changes in biological
diversity and impact on the global carbon cycle." Climatic Change 39(2-3): 317-
336.
Bestelmeyer, B. T., D. Agosti, et al. (2000). Field techniques for the study of ground
dwelling ants: an overview, description, and evaluation. Ants: Standard Methods
for Measuring and Monitoring Biodiversity. D. Agosti, J. D. Majer, L. E. Alonso
and T. R. Schultz. Washington and London, Smithsonian Institution Press.
Bestelmeyer, B. T. and J. A. Wiens (1996). "The effects of land use on the structure of
ground-foraging ant communities in the Argentine Chaco." Ecological
Applications 6(4): 1225-1240.
Bestelmeyer, B. T. and J. A. Wiens (2001). "Ant biodiversity in semiarid landscape
mosaics: The consequences of grazing vs. natural heterogeneity." Ecological
Applications. 11(4): 1123-1140.
Catangui, M. A., B. W. Fuller, et al. (1996). "Abundance, diversity, and spatial
distribution of ants (Hymenoptera: Formicidae) on mixed-grass rangelands treated
with diflubenzuron." Environmental Entomology 25(4): 757-766.
Delabie, J. H. C., B. L. Fisher, et al. (2000). Sampling effort and choice of methods.
Ants: Standard Methods for Measuring and Monitoring Biodiversity. D. Agosti, J.
D. Majer, L. E. Alonso and T. R. Schultz. Washington and London, Smithsonian
Institution Press.
Enquist, C. A. F. (2002). "Predicted regional impacts of climate change on the
geographical distribution and diversity of tropical forests in Costa Rica." Journal
of Biogeography 29(4): 519-534.
Erwin, T. (1989). "Sorting tropical forest canopy samples." Insect Collection News 2(1):
8.
Feener, D. and E. Schupp (1998). "Effect of treefall gaps on the patchiness and species
richness of Neotropical ant assemblages." Oecologia 116(1-2): 191-201.
Fittkau, E. J. and H. Klinge (1973). "On biomass and trophic structure of the central
Amazonian rain forest ecosystem." Biotropica 5(1): 2-14.
Gotelli, N. J. and G. G. Graves (1996). Null models in ecology. Washington, Smithsonian
Institution Press.
Holldobler, B. and E. O. Wilson (1990). The Ants. Cambridge MA, Belknap Press of
Harvarad University Press.
Kaspari, M., L. Alonso, et al. (2000). "Three energy variables predict ant abundance at a
geographical scale." Proceedings of the Royal Society of London, Series B:
Biological Sciences. 267(1442): 485-489.

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Kaspari, M., S. O'Donnell, et al. (2000). "Energy, Density, and Constraints to Species
Richness: Ant Assemblages along a Productivity Gradient." American Naturalist.
155(2): 280-293.
Kaspari, M. and M. D. Weiser (2000). "Ant Activity along Moisture Gradients in a
Neotropical Forest." Biotropica 32(4a): 703-711.
Kremen, C. (1992). "Assessing the Indicator Properties of Species Assemblages for
Natural Areas Monitoring." Ecological Applications 2(2): 203-217.
Kremen, C. (1994). "Biological Inventory Using Target Taxa: A Case Study of the
Butterflies of Madagascar." Ecological Applications 4(3): 407-422.
Majer, J. D. and J. H. C. Delabie (1994). "Comparison of the ant communities of
annually inundated and terra firme forests at Trombetas in the Brazilian Amazon."
Insectes Sociaux 41: 343-359.
Majer, J. D., J. H. C. Delabie, et al. (1997). "Ant litter fauna of forest, forest edges and
adjacent grassland in the Atlantic rain forest region of Bahia, Brazil." Insectes
Sociaux 44(3): 255-266.
Morrison, L. (1998). "The spatiotemporal dynamics of insular ant metapopulations."
Ecology 79(4): 1135-1146.
Nepstad, D. C. (1995). Forest recovery following pasture abandonment in Amazonia:
Canopy seasonality, fire resistance, and ants. Evaluating and monitoring the
health of large-scale ecosystems. C. L. Rapport, C. L. Gaudet and P. Calow.
Berlin and Heidelberg, Springer-Verlag.
Parmesan, C. and G. Yohe (2003). "A globally coherent fingerprint of climate change
impacts across natural systems." Nature 421(6918): 37-42.
Root, T. L. and S. H. Schneider (2002). Climate Change: Overview and Implications for
Wildlife. Wildlife Responses to Climate Change: North American Case Studies.
S. H. Schneider and T. L. Root. Washington, Island Press.
Walther, G. R., E. Post, et al. (2002). "Ecological responses to recent climate change."
Nature 416(6879): 389-395.

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 Appendix I
Equipment list and Suppliers in U.S.
For litter sampling (per transect):
random number table
sighting compass
100 m nylon rope knotted at 10 m intervals
blue flagging
1 1m2 quadrat made of wood or PVC (movable joints; one corner should be openable)
1 machete
2 pair gloves
1 small hand rake
1 small hand trowel
1 watch with seconds or stopwatch
1 litter sifter
10 litter stuff sacks (included with the mini-Winkler apparatus) and some extras
2 waterproof notebooks
pencils
large plastic bag, large enough to carry 10 mini-Winkler samples without compressing them

For pitfall traps (per transect):

soil auger (or sharp edged pipe with diameter of the pitfall cups) to dig pitfall trap holes; must be strong
enough to cut through roots.
10 durable plastic cups, 7-8 cm diameter, 8-10 cm depth, with no ridges inside or at mouth
1 liter 75% Ethanol
5 ml liquid detergent
10 small plastic plates, few cm larger than diameter of cups; used as a roof for the pitfall trap
thin wooden skewers
labels (card stock)
squeeze bottle
forceps
waterproof and Ethanol proof ink pen

For measuring ecological conditions

Kestrel 3000 Pocket Weather Station
Electronic soil probes for pH, temperature, humidity
Small ruler (15 cm)

For processing mini-Winkler litter samples and pitfall trap contents (per transect):
2 pieces nylon rope (5 m)
1 2x3 m plastic sheet (only 2 are needed in total)
10 mini-Winkler apparatus (each mini-Winkler contains a mesh bag, and the external cloth portion) and a
few extra
2 light colored plastic trays
medium width (5 cm) paintbrush
fine (2-3 mm) paintbrush
forceps
10 plastic cups with no ridges inside or at mouth
2 liters 95% EtOH (source alcohol, not denatured EtOH)
squirt bottle
10 vials (25-50 ml) with tight caps
1 large ziploc bag
labels (card stock)
waterproof and EtOH proof ink pen
scissors

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Many of the items on the list are readily available in most cities. Some catalogue or web order suppliers for
items which may be difficult to find in some areas are listed below.

For mini-Winkler apparatus:

Marizete Pereira dos Santos

Km 22 rodovia Ilhéus-Itabuna
cx Postal-07 Cep-45600-000
Ministério da Agricultura - CEPLAC
Centro de Pesquisa do Cacau
SECEN

Residence address:
Rua do Coqueiro nº60
Bairro- Conquista
Ilhéus - Bahia
Cep - 45 650-000
Email: "Paulo Roberto Pires Santos" <[email protected]>

For waterproof pens, waterproof notebooks, specimen processing and identification tools, and any other
general entomological equipment:

BioQuip Products
2321 Gladwick Street
Rancho Dominguez, CA 90220
USA
Telephone: 310-667-8800
Fax: 310-667-8808
e-mail: [email protected]
web: www.bioquip.com

For compass, flagging, waterproof notebooks, pocket weather station, and other general field supplies:

Forestry Suppliers, Inc.

Post Office Box 8397
Jackson Mississippi 39284-8397
USA
Telephone (USA): 800-647-5368
Fax (USA): 800-543-4203
Telephone (International): 601-354-3565
e-mail for international customers: [email protected]
Web: www.forestry-suppliers.com

Ben Meadows Company

PO Box 5277
Janesville WI 53547-5277
USA
Telephone (USA & Canada): 800-241-6401
Telephone (Worldwide): 608-743-8001
Fax (USA & Canada): 800-628-2068
Fax (Worldwide): 608-743-8007
E-mail for international customers: [email protected]
Web: www.benmeadows.com

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 Appendix II
Field data form for leaf litter transects
DATE: (date-Month-year)
NAMES OF COLLECTORS:

ARRAY #:
COORDINATES OF TRANSECT:
STARTING TIME: (hh:mm)
AIR TEMPERATURE AT STARTING TIME:
RELATIVE HUMIDITY AT STARTING TIME:

VEGETATION DESCRIPTION (NUMBER OF LIVE STEMS)

INTERVAL 1-10 m INTERVAL 40-50 m INTERVAL 90-100 m
<1 m HEIGHT

< 2 m HEIGHT

>2 m HEIGHT

>10 cm DBH

CANOPY DESCRIPTION
INTERVAL 1-10 m INTERVAL 40-50 m INTERVAL 90-100 m
APPROXIMATE HEIGHT
(m)
APPROXIMATE HEIGHT
OF CANOPY
EMERGENTS (m)
AMOUNT OF COVER
(CLOSED, PARTIALLY
OPEN, OPEN GAP, ETC)

HABITAT DESCRIPTION (% GROUND COVER OF EACH TYPE)

INTERVAL 1-10 m INTERVAL 40-50 m INTERVAL 90-100 m
SOIL

LIVE VEGETATION
INCLUDING ROOTS
DECOMPOSING WOOD

STONE

LEAF LITTER

LEAF LITTER CONDITIONS

INTERVAL 1-10 m INTERVAL 40-50 m INTERVAL 90-100 m
pH
TEMPERATURE
RELATIVE HUMIDITY
DEPTH (cm)

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 Figures

Figure 1. Placement of ant and termite transects in 1 km2 array. Dotted

lines indicate the divisions among four quadrants of the IMA, within each
of which will be a randomly placed. transect Blue lines indicate transects
which run parallel to each other 3 meters apart.

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 Figure 2. 1m2 quadrat for leaf litter ant sampling. Litter has been
collected in center of square.

Figure 3. Sifting litter (left), and fine litter after having been sifted (right).
The litter sample will be further processed in the lab to extract the ants from it.

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Figure 4. Pitfall trap with mouth flush with the ground (left), and roof of pitfall trap made of plastic plate
and wooden skewers.

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 Figure 5. Transferring litter samples into mini-Winkler mesh sack. Care must be
taken to transfer all the contents of the litter sample, using a paintbrush to sweep
fine particles that fall through back into the mesh.

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 Figure 6. Mini-Winkler extractors (upper). The mesh sack contains the litter sample, and remains
relatively flat as it hangs vertically, centered inside the mini-Winkler. (Lower left photograph is a
view from the top down into the mini-Winkler). The bottom of the mini-Winkler holds a small
cup with a sample label and alcohol (lower right), into which the ants fall when they exit the mesh
sack.

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