bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
1Title
2Look before you jump: jumping spiders discriminate different ants by visual cues
3Sajesh Vijayan1,3, Chethana Casiker2,3 and Divya Uma3*
4
51. School of Biology, Indian Institute of Science Education and Research,
6Thiruvananthapuram, Kerala
72. Ashoka Trust for Research in Ecology and the Environment, Bangalore
83. School of Liberal Studies, Azim Premji University, Bangalore
9*Correspondence: [email protected]
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1 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
24Abstract
25
26Ants, being ubiquitous, aggressive, and top predators, play a predominant role in terrestrial
27ecosystems. Jumping spiders are another prominent invertebrate predator that are present in
28similar habitats as that of ants. Most jumping spiders are thought to avoid ants, yet little is known
29if they discriminate among them. In this study we examined the response of jumping spider
30genus Plexippus towards three different ant species (Oecophylla smaragdina, the weaver ants;
31Camponotus sericeus the golden-back carpenter ants, and Leptogenys processionalis, the
32procession ants). In a behavioral assay that excluded tactile and chemical cues, we tested if
33spiders distinguish the three ants by visual cues alone. We recorded and analysed behaviors such
34as ‘look’, ‘approach’, 'stalk', 'attack', and ‘avoidance’ by spiders towards ants. Our results show
35that the three ants differ in their color, movement and aggressive behavior. Spiders gave 'short
36looks' to live ants, suggesting movement is important in detecting ants. Furthermore, spiders
37gave significantly more ‘long looks’ to procession and golden-back ants compared to weaver
38ants. Spiders approached, stalked and attacked procession ants more compared to weaver ants.
39Numerous jumping spiders and ants overlap in their habitat, and it is advantageous to selectively
40avoid some ants over others. Our results suggests that jumping spiders can indeed distinguish
41ants that co-occur in their habitat by visual cues alone, however, the precise nature of visual cues
42warrants further studies.
43
44Running title: Look before you jump
45Key Words: Jumping spiders, Weaver ant, Procession ant, Carpenter ant, Visual cues bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
46Introduction
47Ants play a prominent role in ecological communities. They are widely distributed, and top
48invertebrate predators in tropical ecosystems (Hölldobler, & Wilson, 1990; Roslin et al., 2017).
49They compete for resources (including prey or nesting areas) with other arthropods, leading to
50non-consumptive effect on arthropod communities (Halaj et al., 1997; Ibarra-Isassi, & Oliveira,
512018). Ants also serve as partners in mutualistic interactions, and their colonies sometimes serve
52as exploitable resources. Because of their ubiquitous presence, aggressive nature and communal
53defense, ants also serve as suitable models for numerous arthropod mimics (McIver, &
54Stonedahl, 1993; Cushing, 1998, 2012)
55
56Since many invertebrate and vertebrate species have overlapping habitats with ants, and many
57even engage in various kinds of interactions with them, it may be advantageous to identify and
58distinguish among different ants. For example, coastal horned lizards selectively consume some
59ants over others and their preferences seem to be influenced by ant size (Suarez et al., 2000).
60Butterflies avoid egg laying on plants frequented by predaceous ants, and this is influenced by
61ant size and shape (Sendoya et al., 2009). Praying mantids avoid ant mimicking spiders because
62of their close resemblance to ants (Ramesh et al., 2016).
63
64Jumping spiders are another top arthropod predator in the terrestrial ecosystem, often found in
65similar habitat as that of ants. Earlier research suggests that many jumping spiders have an innate
66avoidance of ants or subsequently learn to avoid them (Edwards, & Jackson, 1994; Nelson et al.,
672006). However, it is not clear if jumping spiders can distinguish among different kinds of ants.
3 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
68Why might such a discrimination be important? Ants and jumping spiders overlap in their
69habitats, and are often intraguild predators (Okuyama, 2002; Sanders, & Platner, 2006; Sanders
70et al., 2008). Furthermore, some ants may be more aggressive than others: For example, the
71weaver ant (Oecophylla smaragdina) is territorial and highly aggressive towards intruders. In
72such cases, it makes sense for spiders to employ a ‘safety first’ approach, and stay away from
73these ants. However, other ants may not be as aggressive, and jumping spiders could afford to
74forage closer to ant colonies, and sometimes even prey on insects tended by ants (Del-Claro, &
75Oliveira, 2000). In such cases, it may be beneficial for spiders to discriminate one ant from
76another. Jumping spiders may use various subcomponents of the visual signal such as movement,
77color and/or shape to distinguish between ants. Yet, the proximate visual cues these spiders use
78to discriminate different ants are not known. Multimodal cues used by these spiders in the visual,
79chemical or vibrational domain will have important implication for ecology and evolution of ant-
80spider interactions (Clark et al., 2000; Allan, & Elgar, 2001; Nelson, & Jackson, 2006a), &, &.
81The vision of jumping spiders is unique in the animal world, as they have achieved a high degree
82of spatial resolution due to their ‘principal eyes’ (Harland et al., 2012), as well as a wide field of
83view due to ‘secondary eyes’ (Land, 1972; Forster, 1985; Jackson, & Pollard, 1996). Jumping
84spiders use shape, movement, and color to recognize a potential prey: for example, Yllenus
85arenarius rely on the direction of movement, and head-indicating features such as eye spots to
86strike a potential prey (Bartos, & Minias, 2016). Evarcha culicivora, which preys on female
87Anopheles mosquitoes appear to make discriminations based on relative orientation of body parts
88(Dolev, & Nelson, 2014). While movement is more essential than shape for predatory response
89in Phidippus audax (Bednarski et al., 2012) the ant-eating jumping spider Habrocestum pulex
90seems to distinguish its prey even in the absence of motion cues (Li et al., 1996). Both naïve and bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
91field-caught Habronattus pyrrithrix avoided red colored crickets (Taylor et al., 2014), but they
92could also be trained to prefer them (Taylor et al., 2016), demonstrating flexibility in color
93learning during foraging. Although these studies indicate the factors that influence prey
94recognition, cues that salticids use to identify a potential enemy/threat such as an ant have not
95been explicitly examined. Visual predators such as praying mantids avoided red colored ant
96mimicking spiders more than black colored ant mimics (Ramesh et al., 2016). But, it is not
97known if salticids generalize all ants as dangerous or if they can distinguish between different
98species of ants.
99
100The objective of the present study is to examine if jumping spiders can distinguish between three
101species of ants that co-occur in their habitat. We choose three ant species of comparable sizes:
102weaver ants (Oecophylla smaragdina Fabricius 1775), carpenter ants (Camponotus sericeus
103Fabricius 1798) and procession ants (Leptogenys processionalis Jerdon 1851) that differ in color,
104movement and aggression (Table 1). We first establish aggressive behavior, and movement of
105the three ants were indeed different. We then examined if jumping spiders differentiate these ants
106using vision alone by performing behavioral assays where tactile and chemical cues were
107controlled. Finally, by controlling for ant movement, we discuss the importance of color and
108movement cues influencing spider responses.
109
110Methods
111Study species
5 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
112Subadult and adult jumping spiders were collected from in and around Bengaluru, Karnataka
113from Oct 2016 to July 2017. After capture, the spiders were individually housed in plastic boxes
114with holes for ventilation, and a moistened piece of cotton for hydration. Spiders were captured
115two days prior to the behavioral assays. For the assays spiders in the genus Plexippus (Plexippus
116petersi Karsch 1878 and Plexippus paykulli Audouin 1826) were used, as they were
117predominantly found in the same habitat as that of ants. Foragers of all three species of ants and
118houseflies (used as control) were collected on the day of the experiment. Ant and representative
119spider pictures are shown in Fig. 1. All test animals were maintained and released back into their
120natural habitat in compliance with the laws of the country.
121
122Ant aggression towards spiders
123Ant aggression was calculated by restricting an ant (weaver, carpenter, or procession ants n=16
124for each ants) and a spider in a 5 cm3 arena and recording the latency of the ant to bite the spider.
125Ants which took more time to bite (increased latency) were considered to be less aggressive.
126
127Response of spider towards ants
128We examined the response of spiders towards ants, where trials were carried out in an 8cm
129diameter Petri dish. The dish was modified to include an inner transparent ring made with a strip
130of transparent sheet into which the ant was released. The transparent ring neither allowed ants
131nor chemical cues to get into the outer compartment, thus allowing only visual cues to be seen
132from either side of the ring. An individual ant was introduced first, followed by a spider into the
133outer portion, upon which the trial commenced, and was video recorded using a Sony handycam bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
134(HDR-PJ600VE). At the end of 5 minutes, the trial was stopped and the test subjects were
135released back into the storage boxes. Trials were carried out from 9 am to 4 pm, well illuminated
136with a 4.5watt LED lamp. After each trial, the image of the spider was recorded using a Leica
137EZ4E microscope connected to a host computer, later used for identification. Each spider was
138presented with a weaver ant, carpenter ant or a procession ant (n = 29, n = 28 and n = 30 trials
139respectively). Using the same experimental setup, a different set of trials were carried out pairing
140a spider with a housefly to make sure spiders exhibited a typical hunting behavior (n = 25). This
141was to ensure that the set up did not hamper vision-mediated behavior of the spiders. Spiders or
142ants were used only once for all the trials.
143
144Spider responses towards ants/flies were coded as follows: a) Short look: spider orients its
145cephalothorax towards the stimulus for a short amount of time (0-7 seconds) and then leaves b)
146Long look: spider orients its cephalothorax towards the stimulus for longer than 7 seconds and
147then leaves c) Approach: spider moving towards the stimulus, d) Stalk: spider moves slowly
148towards the stimulus, while visually oriented towards it e) Attack: spider pouncing on the wall of
149the outer ring at the stimulus f) Avoid: spider moving away from the stimulus. Short and long
150looks were used by Huang et. al. (2011) to classify jumping spider behavior towards different
151prey. Short looks in our study, on average, lasted for 1.5 secs (median 2.29 secs, range 0 to 7
152secs) across all trials, and followed a beta distribution (Suppl Fig S1). Anything more than 7
153seconds was considered long look, and it ranged from 7 to 112 seconds (average 20.3 secs,
154median 14.1seconds). The proportion of time spent and the frequency of these behaviors in 5
155minutes was measured.
156
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157Difference in ants' movements
158Ant movement was calculated as the amount of time the ants spent moving inside the inner
159transparent ring. The movement scores were obtained from the same trials where spider
160responses (see above) were scored. The presence of spiders outside the ring did not influence ant
161movement.
162
163Responses of spiders in the absence of ant movement
164To control for the movement of ants, in a separate experiment, we created ‘lures’ of ants where
165freshly killed ants were positioned in a life-like manner inside the transparent barrier. Except
166movement cues, color and textural cues were intact. Spider responses (short and long looks)
167towards weaver, procession and carpenter ant lures were compared.
168
169Statistical analyses
170We compared the aggression levels of three ants towards spiders, and movement of ants inside
171the ring by Dunn's-test for multiple comparisons of independent samples with Bonferroni
172correction (Pohlert, 2014). We compared spider responses towards 1) weaver ant and carpenter
173ant 2) weaver ant and procession ant and, 3) carpenter ant and procession ant. The frequency of
174spider responses (approach, attack and avoid) and the proportion of time spent stalking and
175looking towards different species of ants were compared using Kruskal-Wallis test followed by
176post-hoc analysis with Dunn's-test. Behavioral responses of the jumping spiders towards live ants bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
177and ant-lures; and house flies and ants were compared using Mann-Whitney U test. All statistical
178tests were carried out on R version 3.3.2 (R Core Team, 2018).
179
180Results
181Ants differ in their aggression towards spiders
182Our results show that weaver ants, carpenter ants and procession ants differ in their aggression
183towards jumping spiders. Weaver and procession ants were very aggressive, and bit spiders in
184less than a minute after they were introduced as indicated by their average latency time to bite
185the spiders (Table 2). However, carpenter ants were not aggressive: 12 out of 16 trials did not
186elicit any aggressive response from the ants. Carpenter ants are observed to feed on extra floral
187nectaries and tend plant hoppers in the wild, and are known to be less aggressive (Mody, &
188Linsenmair, 2003).
189
190Ants differ in their movement
191Weaver ants and procession ants moved significantly more than carpenter ants when placed
192inside the ring (Table 3). While weaver and procession ants moved continuously, carpenter ants
193moved in a staggered manner, where they would stop and move multiple times. This behavior is
194similar to what is observed in the wild (pers. obs.).
195
196Spiders differentially respond to three ants
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197When controlled for aggression (by putting the ants inside a ring), the jumping spiders
198differentiated the ants by looks. Spiders gave quick short looks (median 2.29 sec) to all the three
199ants (Fig 2. Kruskal-Wallis χ2 = 1.337, df = 2, p-value = 0.5). On the other hand, spiders gave
200significantly longer looks (median 14.1 sec) to golden-backed carpenter and procession ants
201compared to weaver ants (Fig 3. Kruskal-Wallis χ2= 20.029, df= 2, p < 0.001. p values for post
202hoc Dunn’s test: carpenter ant vs weaver ant = 0.001, carpenter ant vs procession ant = 1 and
203procession ant vs weaver ant < 0.001). Furthermore, spiders approached, stalked and even
204attacked procession ants more than weaver ants. But these behaviors were not significantly
205different between carpenter ants and procession ants or carpenter ants and weaver ants (Table 4).
206Since ants were enclosed inside a barrier, they did not impose any threat towards spiders, and
207hence the spider sometimes approached and stalked these ants. When barrier was removed, the
208same spiders avoided these ants (unpublished results).
209Not surprisingly, jumping spiders, readily differentiated ants from the flies, corroborating that
210spiders can indeed differentiate prey from non-prey within our setup. When placed with a fly, all
211spiders exhibited a typical hunting behavior where they stalked (Mann-Whitney U = 665.5, p <
2120.001) and attacked (Mann-Whitney U = 539.5, p< 0.001) the fly significantly more than they
213attacked any of the ants.
214
215 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
216Spiders responses in the absence of ant movement
217In the absence of ant movement, spiders still looked at the ants, but did not differentiate them
218based on looks, suggesting that movement plays a crucial role in differentiating the three ants.
219Dunn's test for looks by spiders towards lures gave non-significant results across all
220comparisons.
221
222Discussion
223Most of the earlier studies suggest that jumping spiders avoid ants, but few have examined the
224response of spiders towards different ants. Our findings indicate that jumping spiders can
225distinguish and respond differentially to different species of ants. Weaver ants, procession ants,
226and carpenter ants differed in their color, aggression and movement, and spiders visually
227discriminated them. In this study we show that both movement and color of the ants influence
228the response of spiders.
229Spiders when placed inside the Petri dish stayed predominantly at the periphery, and responded
230towards ants around 10-20 % of the time during which they differentiated between the ants. Our
231results suggest that movement of the ants is crucial for spiders to detect them. An earlier study
232had shown that jumping spiders innately avoided motionless dead lures of ants by visual cues
233alone (Nelson, & Jackson, 2006). Researchers found that spiders, at the end of 10 hrs, chose to
234stay away from the lures of ants devoid of chemical, movement, and textural cues. Although our
235study is not directly comparable to theirs, in the absence of movement cues the spiders failed to
236differentiate the three ants. Jackson, & Tarsitano (1993) found that out of 11 species of salticids,
237only one ant-specialist spider (Corythalia canosa) attacked motionless ant lures. Movement of an
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238object, be it an enemy, prey or a mate is one of the primary cues that elicit spiders to orient
239towards the signal (Forster, 1977; Jackson, & Pollard, 1996; Bednarski et al., 2012; Bartos, &
240Minias, 2016).
241Our study suggests that while movement of ants elicit detection, other sub-components of visual
242cues (color, brightness, texture and/or behavior) enables spiders to distinguish between them.
243Spiders barely gave long looks to weaver ants compared to golden-back and procession ants.
244Additionally, our preliminary studies (not shown here) with seven other species (Hyllus sp.,
245Menemerus sp., and from five other unidentified species) suggest a similar pattern. We have not
246explicitly tested whether spiders in our study perceive the red, black or black-gold colors of these
247ants, but recent studies have shown that spiders can see red (Zurek et al., 2015), and avoid red
248colored prey (Taylor et al., 2016). Regardless of Plexippus sp can see red, black or black-gold, it
249can nevertheless discriminate the ants.
250While spiders seem to detect ants using short looks, they discriminate ants by giving long looks.
251What may be the significance of such a distinction? First, by giving frequent short looks spiders
252constantly scan their surrounding environment, keeping the target in its field of view. All three
253ants were moving (either continuously or in a staggered manner) inside the transparent ring, thus
254eliciting repeated short looks from spiders. Jumping spiders are known to continuously scan
255novel objects by looking for features like ‘legs’ or ‘head spots’ (Land, 1999; Bartos, & Minias,
2562016). Moreover, jumping spiders in Huang et al.'s (2011) study gave more number of short
257looks to ants and ant mimicking spiders than non-mimetic spiders. Second, quick decisions have
258to be made when an enemy or a potential prey is dangerous or fast moving. For example,
259bumblebees make speedy decisions to avoid potentially dangerous foraging patches (Chittka et bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
260al., 2009). Spiders may give frequent short looks to all ants and make a decision early on if it is a
261potential threat.
262In addition to movement, spiders could use color to distinguish ants. Spiders gave more number
263of long looks to black colored ants; black color could belong to a potential prey or a predator.
264Spiders also approached, stalked and even attacked the black colored procession ants
265significantly more than the red colored weaver ants. This suggests that red colored ants could
266pose a stronger threat than black colored ones. Spiders' response toward black-gold colored
267carpenter ants fell in between their response towards red and black ants. A recent study has
268suggested that gold/yellow color is also aposematic to a wide range of invertebrate and vertebrate
269predators (Pekár et al., 2017). Whether color could play a significant component in
270distinguishing ants remains to be tested.
271If predators with high visual acuity can differentiate ants , do they also respond differently
272towards different ant-mimicking spiders? Interestingly, praying mantids approached spiders that
273mimic black colored carpenter ants significantly more than the spiders that mimic red colored
274weaver ants (Ramesh et al., 2016). That walking like an ant is indeed protective has been shown
275in recent studies (Pekár, & Jarab, 2011; Nelson, & Card, 2016; Shamble et al., 2017). Whether
276visual predators respond to the mimics in the same way, by giving longer looks to some and
277short looks to others needs to be further tested.
278There are over 5000 species of jumping spiders and most of them share habitats with different
279kinds of ants, especially in the tropics. Clearly it is advantageous to differentiate among ants
280rather than to avoid all of them and their mimics. How a spider perceives some ants as
281potentially dangerous is governed by dynamic interaction of multiple sensory modalities and
282contexts. We have shown the importance of visual cues in distinguishing among ants.
13 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
283
284
285Acknowledgement
286This research was funded by a DBT BT/Bio-CARe/04/9809/2013-14, and APU grant to DU.
287Funding for CC and SV came from the DBT grant. We would like to thank Aranya Bagchi who
288helped collect spiders, and Dr. Deepak Deshpande for taking ant pictures.
289Contribution of authors
290CC, SV and DU were involved in project design, data collection and analysis. SV and DU wrote
291the paper.
292
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19 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
402
403Figure legends
404
405Figure 1: Three ant species and the jumping spider used in the study: A. Aggressive fast-
406moving weaver ants B. Aggressive, fast-moving procession ants C. Docile, staggered-moving
407carpenter ants D. Plexippus petersi, jumping spider (Photographs not according to scale)
408
409Figure 2: Proportion of time spider spent giving short looks: Proportion of time spent by
410spiders giving short looks to ants did not vary across the different ants. Kruskal-Wallis χ2 =
4111.3377, df = 2, p = 0.5.
412
413Figure 3: Proportion of time spider spent giving long looks: More time was spent giving long
414looks to procession ant and carpenter ants compared to the weaver ant. Kruskal-Wallis χ2 =
41520.029, df = 2, p < 0.001. Post hoc Dunn's-test: Weaver ant vs Carpenter ant p = 0.001, Weaver
416ant vs Procession ant p < 0.001, Procession ant vs Carpenter ant p = 1df = 2.
417
418
419
420
421 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
422
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429
430Table 1: Ants differed in color, aggressive behavior and movement.
Ant Color Aggression Movement Weaver ant Red High Continuous Procession ant Black High Continuous Carpenter ant Black-gold Low Sporadic
431
432Table 2: Aggression of ants, measured as latency to bite a spider: The more aggressive ants
433take less time to bite a spider. Both procession ants and weaver ants are aggressive compared to
434carpenter ants. Dunn's test for ant aggression: Weaver ant vs Carpenter ant p = 0.007, Weaver ant
435vs Procession ant p = 1 and Procession ant vs Carpenter ant p = 0.001
Ant Latency to attack (sec) Weaver ant (n = 16) 63.32 ± 68.67 Procession ant (n = 16) 59.8 ± 77.6 Carpenter ant (n = 16) 156.9 ± 52.1
21 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
436
437Table 3: Percentage of time ants spent moving in experimental arena: Both weaver ants and
438procession ants move continuously, whereas carpenter ants moved sporadically. Dunn's test for
439percentage of time spent moving by the ants: Weaver ant vs Carpenter ant p < 0.001, Weaver ant
440vs Procession ant p = 1 and Procession ant vs Carpenter ant p < 0.001
Ant Percentage of time spent moving Weaver ant (n = 30) 93.48 ± 9.69 Procession ant (n = 29) 86.41 ± 22.79 Carpenter ant (n = 26) 47.69 ± 38.54 441
442Table 4: Spider responses towards three ants: Spiders differed in their approach, stalk and
443attack towards the three ants (Kruskal-Wallis test: Approach: χ2: 7.45, P=0.02; Stalk: χ2: 10.13,
444P=0.006; Attacks: χ2: 6.35, P= 0.04; Avoid χ2: 0.51, P=0.7). Dunn Post hoc P values are
445presented below.
Weaver ant vs Weaver ant vs Procession ant vs
Behavior Carpenter ant Procession ant Carpenter ant Approach 0.29 0.02 0.20 Stalk 0.185 0.005 0.131 Attack 0.47 0.043 0.167 Avoid 1.000 1.000 1.000
446
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449 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
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462 A B 463
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466 C D 467Figure 1
23 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
468
469
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471
472Figure 2
473
474 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
475
476Figure 3.
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25 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
484
485Supplementary information
486
487Figure S1: Histogram of duration of looks by spiders towards ants: The cut-off for
488differentiating between long and short looks has been established as 7 seconds. The normalized
489durations of short looks (duration less than 7 seconds) follow a beta distribution, with a median
490value of 0.02 (equivalent to 2.29 seconds).
491
492
493
494 bioRxiv preprint doi: https://doi.org/10.1101/349696; this version posted June 18, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
495Table S1: Spider responses towards live and dead ants:
496Spiders spent more time giving short looks to the live ants than their lure. Mann-Whitney U test
497scores comparing long and short looks of spiders towards ants and their lures are given below.
Weaver ant vs Weaver Procession ant vs Carpenter ant vs
Behavior ant lure Procession ant lure Carpenter ant lure Short look U = 390, p<0.001 U = 411, p <0.001 U = 262, p = 0.12 Long look U = 298.5, p = 0.062 U = 313, p = 0.084 U = 296, p = 0.012 498
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