Original Article

Title
Ant communities and their trophobionts shape the incidence of pests and diseases in tropical coffee
agroforestry system

Name(s) of author(s)
Faiz Nashiruddin MUHAMMAD
Akhmad RIZALI
Bambang Tri RAHARDJO

Key words: aphids, coffee berry borrer, dominant, flatids, leaf rust, twig borrer

Address(es) of author(s)
Department of Plant Pests and Diseases, Faculty of Agriculture, University of Brawijaya, Malang
65145, Indonesia. E-mail: arizali@ub.ac.id

Abstract
Ants have very complex role in coffee agroecosystem. Their presence known to have an impact on
biological control in coffee plantation. The purpose of this study was to find out how the ant
community and its trophobiont shape the incidence of pests and diseases on tropical coffee
agroecosystem. Three plots were selected based on age similarity and uniform habitat. Each plot
consisted maximum of 100 coffee trees. Observation was conducted from January to March 2022.
Ant community observation was done three times with once per month frequency using three
methods: visual, tuna bait trap, and live bait trap. Pest and disease observation were also done three
time and once a month. Leaf rust disease, twig borer, and coffee berry borer (CBB) attack intensity
were observed. Five of 21 species of ants were categorized as dominant species. The most common
subfamily found was Dolichoderinae followed by Formicinae. Coffee pest and disease attack were
affected by either ants or their trophobionts hemipteran. Twig borer attack were positively affected
by all hemipteran presence, but negatively affected by D. thoracicus SMITH, 1860 presence. While
CBB were positively affected by flatids and aphids presence, but negatively affected by Camponotus
sp.3 presence. D. thoracicus and Tetramorium spp. were positively affected Coccus viridis GREEN,
1889 that can increase leaf rust disease and twig borer attack intensity. D. thoracicus was also
positively affected flatids and aphids that can increase twig borer and CBB attack intensity. While
Technomyrmex albipes (SMITH, 1861) can indirectly either reduce or increase attack intensity of twig
borer and CBB. Coffee pest and disease attack intensity were also been able to affect each other. In
conclusion, coffee pest and disease incidence were directly and indirectly affected by either ants or
their trophobionts hemipteran.

Introduction
The presence of ants has known to have an impact on biological control that occurs in
agroecosystems (PHILPOTT & ARMBRECHT 2006, WIELGOSS & al. 2013). Ants have a very wide gradient of
roles from cryptic herbivores to specialist predators (DAVIDSON & al. 2003). Ants can have a positive
impact on plant cultivation in agroecosystems if they act as natural enemies or biological control
agents. Plants and ants form a mutualistic symbiosis where ants will provide protection to plants
from herbivorous attacks, while plants will provide shelter, food sources, and breeding grounds for
ant colonies (DIAMÉ & al. 2017). The rate of attack of herbivores on plants not associated with ants
was twice as high as on plants associated with ants (ROSUMEK & al. 2009). Ants can consume large
number of pest, and can also interfere with feeding and oviposition (DRUMMOND & CHOATE 2011). In
addition, ants with their hygienic behavior are also known to suppress the spread of plant pathogens
(OFFENBERG & DAMGAARD 2019). However, the beneficial role of ants is sometimes disturbed by the
dominance of an ant species. In the ant community, the dominant species exhibits aggressive
behavior and can monopolize a food source (STUBLE & al. 2017, LESSARD & al. 2020). Non-dominant
species can coexist with dominant species by foraging in different niches and at different times (VAN
OUDENHOVE & al. 2018).
Besides being beneficial, the presence of an ant species can also be detrimental to plants. Ants with
cryptic herbivore roles will form a trophobiotic relationship with hemipterans (Auchenorryncha and
Sternorryncha) (DELABIE & al. 2001). The relationship of dominant ants with hemipteran species,
some of which are pests, can threaten the sustainability of plant cultivation activities (WIELGOSS & al.
2013). However, ants also choose which species they can become trophobionts (SANCHEZ & al. 2020).
In addition, several dominant ant species are also known to be vectors of the spread of pathogens
(WIELGOSS & al. 2013, RIZALI & al. 2017). The danger from invasive ants is also important. The
presence of invasive ants can damage ecosystems and displace local native species (ANGULO & al.
2022).
Coffee is one of the important plantation commodities in Indonesia besides cocoa and oil palm.
Coffee production in Indonesia reached 762,000 tons in 2020 (BPS 2021). This makes Indonesia
ranked fourth as a coffee exporter in the world (STATISTA 2022). However, coffee cultivation often
faces problems from pests and diseases attack (WALLER & al. 2007). Leaf rust (TALHINHAS & al. 2017),
twig borer (LAN & WINTGENS 2004), and coffee berry borer (VEGA & al. 2009, JOHNSON & al. 2020) are
important pests and diseases of coffee plants in the world that can be found in Indonesia. CBB
attacks in Indonesia range between 10% -20% (WIRYADIPUTRA & al. 2008). Meanwhile, coffee leaf rust
can cause crop losses in the range of 10% -70% (JHA & al. 2014).
Ants in the coffee agroecosystem have a very complex role. VANDERMEER & al. (2010) reported that
ants can also support or interfere with the cultivation of coffee plants directly or indirectly. Ants can
also interact directly or indirectly with pests and diseases of coffee plants. In previous studies, key
ant species from the genus Azteca which are dominant in the South American region can interfere
with CBB colonization (VANDERMEER & al. 2010, GONTHIER & al. 2013, MORRIS & al. 2015). In addition,
ants from the genus Plagiolepis in Kenya can also be predators of X. compactus (EGONYU & al. 2015).
PHILPOTT & ARMBRECHT (2006) also reported that several genera of ants such as Dorimyrmex, Pheidole,
Gnamptogenys, Tetramorium, and Solenopsis can act as predators of coffee plant pests.
The purpose of this study was to find out how the ant community and its trophobiont shape the
incidence of pests and diseases on tropical coffee agroforestry system. Research on the role of ants
in the coffee agroecosystem has never been done in Indonesia. Nearly all publications about the role
of ants as biocontrol of coffee pests and diseases came from the Neotropics and were very limited to
the South East Asian region which is known as one of the largest coffee producing regions in the
world. (PHILPOTT & ARMBRECHT 2006, MORRIS & al. 2018). The results of this study can be used as
recommendations for farmers in the practice of biological control of pests and diseases of coffee
plants.

Materials and methods

Study site: Research was conducted in three coffee plantations at UB Forest, Malang Regency, East
Java, Indonesia (Fig 1). UB Forest is an educational forest managed by University of Brawijaya
covering an area of 554 hectares on the slopes of Mount Arjuno. The altitude range of UB Forest
were from 800 to 1600 meter a.s.l. There were several land use in UB Forest such as coffee
agroforestry, pine forest, natural habitat, annual cropland, and settlement.
Three 30 m x 30 m plot of coffee plants were selected for observation. Each plot consisted maximum
of 100 coffee trees. Plots were selected based on age similarity and uniform habitat. The observed
age range of coffee was 7-10 years. Canopy trees of all plots were pines. The coffee tree in all plots
were also well managed. Fertilizing and pruning were applied once a year.
Ant and trophobiont hemipteran observation: Observation was conducted from January to March
2022. Ant community observation was done three times and once a month with three methods:
visual, tuna bait trap, and live bait trap. Each methods were done in different day. Visual method
was used to determine which species was dominant by observing each tree for 5 minutes. When
observing with tuna bait trap, 200 ml of plastic glass were placed in primary branch. A half
tablespoon of tuna based catfood was put in plastic glass for 2 hours. After that, the ants were
observed which one was dominant and then taken with a brush and then put into 1.5 ml microtube.
While live bait trap was replicated in day and night on 1-day cycle. Two live larvae of Tenebrio
molitor LINNAEUS, 1758 were put in plastic glass for 1 hour. Then, the ants were also observed which
one was dominant and then put it in 1.5 ml microtube. The dominant ants were taken as many as
three individuals, while the non-dominant ants were taken 1 individual per species. Ants were
identified to genera using (NAZARETTA & al. 2021) and then separated to morphospecies based on
external morphology (LATTKE 2000). The purpose of this kind of observation was so that the
dominant ant colony is not disturbed in the future observation.
Hemipteran observation was only done with visual. Each tree were observed in 5 minutes. Every
hemipteran that associate with ants (trophobiosis) were directly recorded and photographed. There
were 5 species of hemipteran that associated with ants: two species of flatids, two species of aphid,
and green scale Coccus viridis GREEN, 1889. The observeation were also done three times, once a
month.
Pest and disease incidence observation: Pest and disease observation were done three time and
once a month. Leaf rust disease (Hemileia vastatrix BERK & BROOME, 1969), twig borer (Xylosandrus
compactus (EICHHOFF, 1875)), and CBB/coffee berry borer (Hypothenemus hampei (FERRARI, 1867))
attack intensity were observed. The incidence of each pest and disease were count based on the
intensity. Four branch samples were determined based on point of compass (North, West, South,
East). The intensity of leaf rust was calculated by comparing the number of symptomatic leaves with
the total number of leaves on the sample branches. The intensity of twig borer attack was calculated
by comparing the number of wilted twigs with the total number of twigs in the sample branches.
While the intensity of CBB attack was calculated by comparing the number of hollow fruits with the
total fruit in the sample branches. The intensity value was obtained by dividing the number of
symptomatic plant parts by the total plant parts, then multiplied by 100.
Data analysis: All data were organized as database using Google Spreadsheet first. Ant and
hemipteran observation results from three times observation were combined to get occurences and
present-absent data. While intensity value of pest and disease were averaged from three times
observations. Differences in attack intensity of each pest and disease of coffee plants were analyzed
by Analysis of Variance (ANOVA). If significant difference was found, then Tukey's post-hoc test were
conducted. Before conducting ANOVA, the data was tested for normality with the Shapiro-Wilk Test.
If the data was not normally distributed, then the data was transformed using log(x). The
relationship between each dominant ants and hemipteran presence were analyzed using
Generalized Linear Model (GLM) (ZUUR & al. 2009) with quasipoisson distribution to account for
overdispersion. GLM was also used to test the relationship of presence/absence of ant and
hemipteran to coffee pest and disease attack intensity. Pearson Product Moment Correlation Test
were also conducted to analyzed interaction between each pest and disease attack intensity. All
analysis were conducted using R Statistic version 4.2.0 with additional package vegan (R CORE TEAM
2022).

Results
Ant and trophobiont hemiptera community: The total ants found at the study site were five
subfamilies, 13 genera, and 21 mor-phospecies (Tab 1). The most common subfamily found was
Dolichoderinae (46.5% of total tree) followed by Formicinae (23.7%), Myrmicinae (17.6%),
Pseudomyrmicinae (10.1%) and Ponerinae (1.8%). In the comparison of each plot, Dolichoderinae
ants was also the most commonly found. At the species level, Dolichoderus thoracicus SMITH, 1860
and Technomyrmex albipes (SMITH, 1861) were the most abundant species. The highest occurrence
of ant species in plots A and C was D. thoracicus. While in plot B, the highest species occurrence was
T. albipes. Eleven species were shared on all three plots.
Five of 21 ant were categorized as dominant ant in coffee canopy. They have ability to build
connected nest (polidomous nest), have huge number of workers, and dominant in bait trap. All
dominant ant can be found in all plots except Crematogaster sp.1. Other ant that have similar
characteristic with dominant ant were Polyrachis armata (LE GUILLOUU, 1842), Tetramorium spp., and
Pheidole sp.1. They were found in huge number of worker but their nest were not in coffee canopy.
P. armata built their nest in pine trees canopy, while Tetramorium and Pheidole ants built their nest
under the ground near coffee main stem. P. armata and Tetramorium spp. were found foraging in
canopy but they didn’t dominate the bait trap. While Pheidole sp.1 rarely found foraging in canopy
but they can dominate the bait trap.
Each dominant species have different type of nest in canopy. Dolichoderus thoracicus only built their
nest by attaching some dried coffee leaves. Technomyrmex albipes built their nest on the bended
leaf, under the bark, and lichen. Camponotus sp.3 only built their nest on the lower leaf surface.
They attached some soil to build their nest. Crematogaster sp.1 also built the nest by attaching some
soils on the primary branch and twig. Their center nest was in the primary branch, while many
secondary nests were built in the twig segment. Crematogaster sp.1 nest size was also bigger than
other dominant ant nest. Tetraponera sp.1 was very different than the other dominant ants. They
used the tunnels in the twig that were made by coffee twig borer. All dominant ants have ability to
explore entire canopy.
Due to the difference type of nest, dominant species can share the space to other dominant species
(Fig 2). One tree can contain up to four dominant species. Some trees were only occupied by non-
dominant species and a few trees were not occupied by ants.
Five species of trophobiont hemipteran were found during sampling (Tab 2). The most abundant
trophobiont hemipteran was Flatidae sp.1 followed by green scale Coccus viridis (Hemiptera:
Coccidae). Flatidae sp.1 was very abundant in plot A, while C. viridis was most common in plot B and
C. Flatids and C. viridis can be found at twig and leaf stalk (petiole). While aphids can be found only
at lower leaf surface.
All thropobiont hemipteran presence were affected by dominant ants (Tab 3). Coccus viridis were
positively affected by Dolichoderus thoracicus and Tetramorium spp.. Flatids presence were
positively affected by D. thoracicus, but negatively affected by T. albipes. While aphids were
positively affected by D. thoracicus and Technomyrmex albipes.
Effect of ant and hemipteran on coffee pest and disease attack: The highest attack intensity was
caused by twig borer (16.08%), followed by leaf rust (13.34%) and CBB (11.44%). When compared,
there was no difference in the intensity of attack by leaf rust (F 2.253 = 0.033; P = 0.968) and twig borer
(F2.253 = 1.996; P = 0.138) in each field. Whereas in CBB, the attack intensity was different in each field
(F1.161 = 8.828; P <0.001), and the highest attack occurred in plot B.
Coffee pest and disease incidence were affected by either ants or their trophobiont hemipteran (Tab
4). Twig borer and CBB attack intensity were affected by both ants and hemipteran. While leaf rust
disease attack were only affected by hemipteran. Leaf rust disease intensity were only positively
affected by the presence of Coccus viridis. Twig borer attack were positively affected by all
hemipteran presence, but negatively affected by Dolichoderus thoracicus presence. While CBB were
positively affected by flatids and aphids presence, but negatively affected by Camponotus sp.3
presence.
Interaction between ants, hemipteran, and coffee pest disease: The connection between ants,
hemipteran, and coffee pest and disease has formed a complex interaction (Fig 3). Ants presence can
affect directly and indirectly to coffee pest and disease. Only two of seven ants that analysed were
known to directly reduce attack intensity of coffee pest. While three of seven ants were known to
indirectly affect the attack intensity of coffee pest and disease.
Dolichoderus thoracicus and Tetramorium spp. were positively affected Coccus viridis that can
increase leaf rust disease and twig borer attack intensity. D. thoracicus was also positively affected
flatids and aphids that can increase twig borer and CBB attack intensity. While Technomyrmex
albipes can indirectly either reduce or increase attack intensity of twig borer and CBB. Coffee pest
and disease attack intensity were also been able to affect each other. Positive correlations were also
found in the interaction of each coffee pest and disease. Based on correlation analysis result,
positive correlation was found between leaf rust disease with twig borer (r = 0.311; P < 0.001), leaf
rust disease with CBB (r = 0.189; P = 0.015), and twig borer with CBB (r = 0.302; P < 0.001).

Discussion
The dominant ants that was found during observation belonged to the functional groups Dominant
Dolichoderinae (Dolichoderus thoracicus), Opportunist (Technomyrmex albipes), Subordinate
Camponotini (Camponotus sp.3), Generalized Myrmicinae (Crematogaster sp.1, Pheidole spp.,
Tetramorium spp.), and Specialist (Tetraponera sp.1) based on ANDERSEN (2000) and BHARTI & al.
(2013). The only invasive species found was T. albipes (PFEIFFER & al. 2008). Although no other
invasive ants such as Anoplolepis gracilipes (SMITH, 1857) and Paratrechina longicornis (SMITH, 1802)
were found, the presence of Dominant Dolichoderinae and some Generalized Myrmicinae proved
that stress and disturbance by invasive ant species in the observed area was quite high (MCGLYNN
1999).
This research was evidence that ants can act as biological controllers for pests and diseases that
attack coffee plants. At this study, two species of ants (Dolichoderus thoracicus and Camponotus
sp.3) were able to control two species of ambrosia beetle (Hypothenemus hampei as CBB and
Xylosandrus compactus as twig borer) which act as pests on coffee plants. Some ant species in the
research of PHILPOTT & ARMBRECHT (2006) are known to act as biological controllers in coffee plants.
However, that study was only conducted in the Africa (Afrotropical) and American (Neotropical)
regions. Tetramorium spp. and Pheidole spp. were included in the list as CBB predators, but in this
study the presence of these two genera did not affect the intensity of coffee pests and diseases. In
CBB attacks, these results prove that the presence of ants can reduce CBB attacks. This is in
accordance with the experiment by GONTHIER & al. (2013) whose results proved that some ant
species can prevent CBB from colonizing. In addition, in previous studies, ants can also reduce the
attack of Xylosandrus compactus as coffee twig borer. GIANNETTI & al. (2022) research also proved
that female X. compactus avoids twigs containing foraging ants as hosts.
Regarding the relationship between ants and hemipteran, ants chose which hemipteran species that
can become their trophobiont. In previous studies, an antagonistic mechanism was found between
ants and the hemipteran. The results of the study by BILLICK & al. (2007) showed that Formica
obscuripes FOREL, 2006 can move aphids and reduce feeding intensity. Ants also have a preference
for certain hemipterans. The results of the study of (SANCHEZ & al. 2020) showed that the dominant
species Lasius grandis FOREL, 2006 could reduce the population of Cacopsylla sp. in the pear plant.
However, on the other hand, ants can make some species of hemipteran into their trophobiont. The
ants provide protection to the hemipteran. In return, the hemipteran provide honeydew to the ants.
According to (KATAYAMA & al. 2013), ants switch from extrafloral nectar (EFN) to aphids honeydew
because the aphids provide more rewards for the ants. Even ants can turn to prey on aphids when
an artificial sugar solution has been provided in the field (PÉREZ-RODRÍGUEZ & al. 2021). Ants need
honeydew, EFN or sugar to meet their carbohydrate needs (RUDOLPH & PALMER 2013). In coffee
plants, Coccus viridis has been recorded as the trophobiont of many ant species (BACH 1991, PHILPOTT
& ARMBRECHT 2006). In the Neotropics, Azteca instabilis interferes the predation process of the
coccinelid beetle Azya orbigera on C. viridis (PERFECTO & al. 2014).
The positive correlations between leaf rust, twig borer, and CBB is evidence that pest and disease
attacks can reduce plant resistance. Weak plant resistance causes plants to be easily attacked by
pests and diseases. Pathogenic fungi can take up nutrients and interfere with plant physiological
processes (DIVON & FLUHR 2007). HONORATO JÚNIOR & al. (2015) also explained that Hemileia vastatrix
leaf rust attack can reduce the rate of photosynthesis of coffee plants. In addition, the twig-boring
ambrosia beetle is also associated with several fungal species that can grow in boreholes [3]. Several
types of microbes known to be associated with ambrosia include: Aspergillus spp., Penicillium spp.,
Trichoderma spp., Fusarium spp., Acremonium spp., and Gliocladium spp. (TARNO & al. 2016).
Fusarium and Aspergillus were also known to be found in coffee fruit infected with CBB (ALVES DA
SILVA & al. 2020). According to HUBER & al. (2011), pest and disease attacks can reduce the supply of
nutrients for plants, resulting in a decrease in the level of plant resistance. Another reason for the
positive correlation between pests and diseases was proven by (DE BOBADILLA & al. 2022) who
explained that a plant's resistance can be determined by sequential attacks from pests. The positive
correlation between hemipteran trophobion also supports the presence of sequential attacks on
coffee plants. Ants, which mostly have a positive correlation with hemipteran also indirectly
contribute to pest and disease attacks. Only T. albipes showed a negative correlation with one of the
hemipteran trophobiont.
In conclusion, in an ant community, there were dominant and non-dominant species. The presence
of ants and their trophobes affected the incidence of coffee pests and diseases. Ants had either
positive or negative impact on the incidence of coffee pests and diseases according to previous
research by (VANDERMEER & al. 2010). Each ant species has a different impact on pests and diseases of
coffee plants, directly or indirectly. All of the Hemiptera trophobiont only had negative impact on
the incidence of coffee pests and diseases. The absence of the antagonistic fungus Akanthomyces
lecanii (ZIMMER, 1898) associated with Coccus viridis (JACKSON & al. 2012) made the role of C. viridis as
an indirect biological control of coffee leaf rust absent.

Acknowledgments
Thanks to our field assistants, Reza Diaz F. R., Bagas Agung P., Enjang Sutrisno, Minhajul Qowim, Mei
Irawan, Bayu Budi P., and Adilla Haqi. Thanks to the owner of coffee agroforestry, Pak Ngateri, Pak
Suyadi, and Pak Riyadi. Thanks also to the staff of the UB Forest, especially to Pak Yohannes and Pak
Bambang for allowing us to do research in UB Forest area.