Behavioral Manipulation of the Spider Macrophyes Pacoti (Araneae: Anyphaenidae) by the Araneopathogenic Fungus Gibellula Sp

Canadian Journal of Zoology

Behavioral manipulation of the spider Macrophyes pacoti (Araneae: Anyphaenidae) by the araneopathogenic fungus Gibellula sp. (Hypocreales: Cordycipitaceae)

Journal: Canadian Journal of Zoology

Manuscript ID cjz-2020-0232.R2

Manuscript Type: Article

Date Submitted by the 14-Dec-2020 Author:

Complete List of Authors: Arruda, Italo; Universidade Federal do Ceará, Centro de Ciências, Departamento de Biologia; Universidade da Integração Internacional da Lusofonia Afro-Brasileira, Laboratório de Ecologia e Evolução, Instituto de CiênciasDraft Exatas e da Natureza Villanueva-Bonilla, German; Universidade Estadual de Campinas Faustino, Marcio; Universidade da Integração Internacional da Lusofonia Afro-Brasileira, Instituto de Ciências Exatas e da Natureza Sobczak, Jullyana; Universidade Federal do Ceará, Centro de Ciências, Departamento de Biologia Sobczak, Jober; Universidade da Integração Internacional da Lusofonia Afro-Brasileira, Instituto de Ciências Exatas e da Natureza

Is your manuscript invited for consideration in a Special Not applicable (regular submission) Issue?:

Host Manipulation, Entomopathogenic fungi, Massif of Baturité, ghost Keyword: spider, Macrophyes pacoti, Gibellula

Behavioral manipulation of the spider Macrophyes pacoti (Araneae:

Anyphaenidae) by the araneopathogenic fungus Gibellula sp.

(Hypocreales: Cordycipitaceae)

I.D.P. Arruda

Universidade Federal do Ceará, Centro de Ciências, Departamento de Biologia,

Programa de Pós-Graduação em Ecologia e Recursos Naturais, Fortaleza, CE, Brazil.

[email protected]

G.A. Villanueva-Bonilla

Programa de Pós-graduação em Biologia Animal, Instituto de Biologia, Universidade Estadual de Campinas, CP 6109, CEPDraft 13083-970, Campinas, SP, Brazil. [email protected]

M.L. Faustino

Universidade da Integração Internacional da Lusofonia Afro-Brasileira – UNILAB,

Instituto de Ciências Exatas e da Natureza, Laboratório de Ecologia e Evolução, R. José

Franco de Oliveira, s/n, Redenção, Ceará, 62790-972, Acarape, CE, Brazil.

[email protected]

J.C.M.S. Moura-Sobczak

Universidade Federal do Ceará, Centro de Ciências, Departamento de Biologia,

Programa de Pós-Graduação em Ecologia e Recursos Naturais, Fortaleza, CE, Brazil.

[email protected]

1 © The Author(s) or their Institution(s) Canadian Journal of Zoology Page 2 of 30

J.F. Sobczak

Laboratório de Ecologia e Evolução, Instituto de Ciências Exatas e da Natureza,

Universidade da Integração Internacional da Lusofonia Afro Brasileira, Rodovia CE-

060, Km 51, s/n, 62785-000, Acarape, CE, Brazil. [email protected]

Corresponding author: German Antonio Villanueva-Bonilla. Instituto de Biologia,

Universidade Estadual de Campinas, CP 6109, CEP 13083-970, Campinas, SP, Brazil.

Phone number: +55 (19) 981937682. E-mail: [email protected].

Draft

2 © The Author(s) or their Institution(s) Page 3 of 30 Canadian Journal of Zoology

I.D.P. Arruda, G.A. Villanueva-Bonilla, M.L. Faustino, J.C.M.S. Moura-Sobczak, J.F.

Sobczak

Behavioral manipulation of the spider Macrophyes pacoti (Araneae:

Anyphaenidae) by the araneopathogenic fungus Gibellula sp. (Hypocreales:

Cordycipitaceae)

Abstract

Host manipulation has already been documented in several distinct host-parasite

associations, covering all major phyla of living organisms. While in animals we know

that several species have the ability to manipulate their hosts for the benefit of the parasite, in arthropopathogenic fungi there Draft is very little knowledge about possible behavioral manipulation. We report for the first time the interaction between the araneopathogenic

fungus Gibellula sp. Cavara and the spider Macrophyes pacoti Brescovit, Oliveira,

Sobczak & Sobczak, 2019 (Anyphaenidae) in addition to investigating the potential

change in behavior of spiders infected by the parasitic fungus. We also investigated

whether the rainfall regime influences the abundance of infected spiders and the

parasitism rate by the araneopathogenic fungus. Our results corroborated our hypothesis

that the fungus induces vertical segregation in the spider population, inducing infected

spiders to be at higher heights than uninfected ones. Dead infected spiders were found in

a stretched position that probably helps in fixing the carcass on the leaves by increasing

the contact surface between the host and the substrate. Our results also confirm the

positive relationship between the rainy season and the greater number of parasitic spiders

and the parasitism rate.

Keywords: Host Manipulation, Entomopathogenic fungi, Massif of Baturité. ghost

spider, Macrophyes pacoti, Gibellula.

3 © The Author(s) or their Institution(s) Canadian Journal of Zoology Page 4 of 30

INTRODUCTION

Parasites are of great importance for the functioning of communities and ecosystems, and can influence the population dynamics of their hosts, as well as act on the persistence of these species through their pathogenic effects (Lefèvre et al. 2009;

Hatcher et al. 2012). Some species of parasites have the ability to manipulate the behavior of their hosts to survive, transmit, and complete their life cycle (Moore 1984;

Poulin et al.1994). Host manipulation has already been documented in several distinct host-parasite associations, covering all major phyla of living organisms (Moore 2002).

In animals, we know that some species have the evolutionary characteristic of an extended phenotype, which is the ability to manipulate their host in a way that benefits the parasite. For example, moth caterpillars Spodoptera exigua Hübner, 1808, normally live hidden in the soil, but when infectedDraft with baclovirus they rise in the treetops where they die, melt and release their virions that will infect new caterpillars (Van Houte et al.

2014). The fish Fundulus parvipinnis Girard, 1854, known as “killifish” infected by the trematode Euhaplorchis californiensis Martin, 1950 has its behavior altered where its movements become more conspicuous and makes it more visible. This altered behavior makes them more easily depredated upon, favoring the trophic transmission of the trematodes from the intermediate host, fish, to the definitive host, bird (Lafferty and

Morris 1996).

Entomopathogenic fungi also stand out as a group of parasites that manipulate the behavior of their hosts, with some examples already studied including flies (Elya et al. 2018), beetles (Steinkraus et al. 2017), and, mainly ants, which are popularly known as “zombie ants” (Andersen et al. 2009; Pontoppidan et al. 2009; Hughes et al. 2011; De

Bekker 2019; Kurze et al. 2020). In the literature, two types of manipulative effects can be observed in the hosts, (1) induce the host to select a different habitat which will

4 © The Author(s) or their Institution(s) Page 5 of 30 Canadian Journal of Zoology

benefit the spore dispersion (e.g. Maitland 1994) and (2) modify the final posture of the

host to increase the surface of contact and adhesion to the substrate (Hughes et al.

2011).

There are species of fungi that infect some arachnids such as mites (Trandem et

al. 2015), harvestmen (Barbosa et al. 2015; Santamaria et al. 2017) and spiders, the

latter being called araneopathogenic fungi (Evans and Samson 1987; Evans 2013;

Hughes et al. 2016). Records of spiders being infected with fungi are abundant, but so

far none of these studies have tested the possibility of behavioral manipulation of

infected spiders (Evans and Samson, 1987; Evans 2013; Costa 2014; Hughes et al.

2016; Brescovit et al. 2019). Among the araneopathogenic fungi, some are considered

specific to spiders, the best known being grouped in the genera Torrubiella Boud.,

which comprises the sexual teleomorphicDraft stage, and Gibellula Cavara, which represent

the asexual anamorphic stage, both included in the Cordycipitaceae family (Johnson et

al. 2009). Currently these genera of fungi comprise 23 and 80 species described

respectively with worldwide distribution (Kobayasi and Shimizu 1982; Kirk 2020).

Several of the host spider species are adult individuals from the Salticidae family

(Evans and Samson 1987; Samson and Evans 1992), but in many records the material

collected describes the host as an “unidentified spider” (Johnson et al. 2009) which

limits the degree of knowledge about possible host specificity of the parasitic fungus.

Recently, Brescovit et al. (2019) described a new species of spider in the family

Anyphaenidae, and recorded the interaction between this species and the

araneopathogenic fungus Gibellula sp. but without any details about the possible

behavioral manipulation. The Anyphaenidae family is represented by small to medium

wandering hunting spiders. The group is relatively uniform and well defined,

morphologically and geographically (Ramírez 2003). Currently, this family is composed

5 © The Author(s) or their Institution(s) Canadian Journal of Zoology Page 6 of 30

of 56 genera and 572 species described (World Spider Catalog 2020). The greatest diversity of Anyphaenidae occurs in the New World, especially in South America, with

29 endemic genera (World Spider Catalog 2020). Interactions between Anyphaenidae and araneopathogenic fungi are scarce due to the difficulty of correctly identifying the host spider because sometimes it is completely covered by the parasitic fungus (Hughes et al. 2016). However, a few records reported that Anyphaenidae represents one of the most frequent spider families parasitized by Gibellula in the inventories, with the species Iguarima sensoria Keyserling, 1891 being the most commonly identified (Costa

2014; Hughes et al. 2016).

In one of the Atlantic Forest remnants of the state of Ceará-Brazil, we registered several individuals of the species Macrophyes pacoti (Anyphaenidae) at different heights on tree trunks, some of whichDraft were infected with the fungus Gibellula sp. Thus, the objectives of the present study were: (1) to describe the natural history of the interaction of parasitism between the spider M. pacoti and the fungus Gibellula sp., (2) to investigate potential change in the behavior of spiders infected by the araneopathogenic fungus, (3) assess whether the abundance of spiders parasitized by the fungus varies throughout the year, and (4) evaluate whether the monthly rainfall regime influences the abundance of infected spiders and the parasitic rate of the araneopathogenic fungus. Abundance is defined as the monthly registered amount of spiders parasitized by the Gibellula fungus and the parasitism rate is the percentage of spiders parasitized per month taking into account the total number of spiders registered

(parasitized or not). We expect that infected spiders will have their behavior manipulated and that they occur on average at higher heights in the trunks when compared to the height of spiders not infected by the fungus. We also expect that the abundance of infected spiders and the rate of parasitism will be higher in the period of

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the year with the highest rainfall; high humidity benefits the development of the

araneopathogenic fungus.

MATERIAL AND METHODS

Study area

We conducted the study in a forest in the Environmental Protection Area (APA)

of Serra de Baturité, Ceará, Brazil. In this area, the typical vegetation of the Atlantic

Forest predominates (Oliveira and Araújo 2007), being classified as Wet Forest and Dry

Forest of Cristalino (Moro et al. 2015), also called, respectively, Montana Evergreen

Seasonal Forest and Montana Semideciduous Seasonal Forest (IBGE 2012) with

altitudes ranging from 800 to 890 m asl. The climate of the region is sub-humid tropical.

The average annual temperature variesDraft between 19 and 22 °C and the average annual

precipitation is 1500 mm (Semace 2010) (Fig. 1).

In the APA region of Serra de Baturité, we selected an area known as “Mata do

Purgatório” (Fig.2) (4°13' 21.10'' S, 38°53' 35.80'' W), located at “Sítio São Luís”, in the

municipality of Pacoti-Ceará.

Fungus-spider interaction and behavioral manipulation

In the study area, we demarcated five sample plots of 10 x 10 m (100 m²) each

100 m apart. In these plots, monthly and for a year (from September 2018 to August

2019), we conducted active and detailed searches of M. pacoti infected or not by fungi

Gibellula sp. on the trunk and leaves of the trees from the ground up to 2.5 m in height.

The record was performed up to 2.5 m in height based on the methodology of previous

studies done with the ant-fungus interaction (Araújo et al. 2015; Cardoso-Neto et al.

2015). For each spider collected, we recorded whether the individual was infected or

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not, in addition to recording the height in relation to the ground. To distinguish infected from uninfected spiders, we observed the growth of hyphae in the host's body. If the observed spider did not show growth of hyphae of the fungus, then it was counted as not infected. Additionally, we looked in the field where the parasitic spiders were specifically (trunk or leaves), in parasitic spiders, we observed where the growth of hyphae in the host's body began. We also looked in the field for the spider web shelter.

The collections were carried out during the day (from 10 am to 3 pm), with a sampling effort of five people for approximately one hour for each plot in each month, totaling 60 hours of observations during the period of one year.

In the same plots, we carried out focal observations of the behavior of infected and uninfected spiders as well as the micro-habitat where they were usually observed for later comparisons. The observationsDraft totaled 35 hours of observation, being 25 daytime and 10 nighttime.

Subsequently, to test whether spiders infected by fungi are induced to vertical segregation in the vegetation, we compared the average height between spiders infected and not infected by the fungus using generalized linear models (GLM) after having tested the actual data for not-normality using the Shapiro-Wilk test. For the test, the mean values of height in relation to the ground (cm) of infected and uninfected spiders were considered as the response variable and the condition of parasitism (infected or not infected) was considered the explanatory variable.

Frequency of parasitic fungus-spider interaction

In the plots demarcated in the field, we accounted for individuals of M. pacoti infected or not by fungi Gibellula sp. To estimate the rate of parasitism "RT" in each month of sampling, we divide the number of infected spiders “NIS” among the total

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number of registered spiders (infected or not) “TNS”. Therefore, the value obtained was

multiplied by 100 to know the percentage of parasitism per month:

Subsequently, to find out whether the abundance of infected spiders differs

between the sampled months, we used generalized linear models (GLM) with a Poisson

distribution (count). In addition, to check for marked peaks of infected spiders

throughout the year, we performed circular histograms and circular statistics using the

Rayleigh uniformity test, once the circular normality of the data was tested in the Oriana

circular statistics program version 4.0 (Kovach 2011; Morellato et al. 2010).

Additionally, we use the length of the "vector r" generated by the Oriana software

which indicates how strong the peak of parasitic spiders can be throughout the year.

Synchrony between abundance ofDraft parasitized spiders, parasitism rate and

precipitation

To verify if the monthly variation in the abundance of spiders parasitized by the

fungus and the rate of parasitism are related to the monthly precipitation variation, we

used the Pearson and Spearman correlation test respectively, once the normality of the

data was tested using the Shapiro-Wilk test. The pluviometric data used in this study

were collected by the pluviometric station (Code 105 of Funceme - Cearense

Foundation of Meteorology and Water Resources) in the city of Pacoti-CE.

For all analyzes, except for circular statistics, we used the free software R (R

Core Team 2019) with the “ggpubr” and “DescTools” packages (Kassambara 2019;

Signorell et al. 2019).

RESULTS

Fungus-spider interaction and behavioral manipulation

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In total, we registered 1170 specimens of M. pacoti of which 357 individuals were parasitized by the fungus Gibellula sp. (Fig. 3). All infected spiders were found dead with the ventral part of the body facing the leaf and the dorsal part of the body facing the ground, always on the abaxial surface of the leaves (Fig. 3A-B). The parasite covers the body of the spider partially or integrally with its reproductive structures (Fig.

3C-D), starting from the abdomen and going to the spider's cephalothorax. The mycelium of the parasitic fungus is multiseptate, warty type, which joins in a tangle of hyphae giving rise to synnemata. Synnemata are spread throughout the host's body, with a long and robust stem of a golden-yellow to orange color with white conidiophores formed only at the apex (Fig. 3C - D). After the death of the spiders, the fungus uses the spider's body as a substrate for growth and development of spore-forming structures that will infect new individuals. Draft

Infected spiders are attached to the leaves of plants through fungal hyphae that are projected out of the host's body. Even if the host is dead, its body remains attached to the leaf, keeping it in a stretched position (Fig. 3), where its abdomen and legs maintain this support. This stretched leg position is not observed in dead non-parasitized spiders

In the behavioral observations, we found that the uninfected individuals of M. pacoti are more active at night, and during the day they are in silk shelters that are located in the abaxial part of the leaves of the trees (Fig. 4). On these same leaves we occasionally register infected and uninfected spiders. These silk shelters are usually built in the central region of the leaf, close to the central rib (Fig. 4).

during the day uninfected spiders spend most of the time inside the shelter, with the ventral part facing the ground, leaving only at times when other arthropods come close, such as ants, butterflies, mites and other spiders. During the night, when they are

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most active, we observed several individuals raising their pedipalps when some

dipterans approached the leaf where the spider was. we observed that all spiders

infected by the fungus were always outside the silk shelter and with their legs fully

stretched, a position that we did not observe in uninfected spiders (Fig. 4).

Behavioral manipulation

The infected and uninfected spiders registered occurred at an average height of

74.55 cm ± 37.58 cm SD, ranging from 5 to 240 cm. The average height in relation to

the ground of the infected spiders (99.17 ± 39.61 SD) was significantly higher when

compared to the average height of the uninfected spiders (74.27 ± 35.90 SD) (GLM: t =

101.1628; p = <0.0001) (Fig. 5). Draft

Frequency of parasitic fungus-spider interaction

The abundance of infected M. pacoti individuals did not vary significantly over

the year (GLM: Z = -0.26; p = 0.78). However, we note that there is a peak of

abundance between the months of March and May (Rayleigh Test: z = 14.513; p =

<0.0001) represented by the red cross line outside the circle (Fig. 6) (Table 1), but this

peak was not very strong due to the low value of the length of the vector r (0.083).

Association of rainfall with parasitism

We recorded a positive relationship between the abundance of infected spiders

and monthly precipitation (R = 0.57, p = 0.05) (Fig. 7A), and also between the

parasitism rate and monthly precipitation (R = 0.78, p = 0.0041) (Fig. 7B).

DISCUSSION

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Our study is one of the first studies to present a case of change in the behavior of spiders induced by araneopathogenic fungi, in addition to showing how this host- parasite interaction is influenced by the degree of rainfall. The study provides three key results: (1) First, we found a clear pattern in which spiders infected by the fungus were at higher heights when compared with the values of uninfected spiders, corroborating our hypothesis of behavioral manipulation by the fungus parasite. (2) Second, this study provides evidence that individuals of M. pacoti infected by Gibellula sp. are found in different positions when comparing with uninfected individuals. (3) Third, our results also support us to confirm our hypothesis that the abundance of infected spiders, as well as the parasitism rate, is positively correlated with the monthly precipitation of the study site.

The manipulation of the hostDraft by the parasite or parasitoid is a behavior widely distributed among different taxa, such as viruses, bacteria, fungi and arthropods (Moore

2013; Hughes et al. 2016). Our results show that M. pacoti spiders infected by the fungus are found in tree leaves at higher heights than uninfected spiders. This vertical displacement of infected spiders is likely to benefit fungal dispersion and transmission, one of the two general cases that benefit the parasite (Poulin et al. 1994). Although the advantage of vertical displacement for the fungus has not been tested experimentally, similar results have been recorded in ants of the genus Camponotus Mayr, 1861 that are infected with entomopathogenic fungi of the Ophiocordyceps unilaterallis complex sensu lato (Andersen et al. 2009; Araujo et al. 2015). In this ant-fungus system, hosts are usually found on substrates up to 2 m in height and some species of the parasitic fungus exhibit specific shapes to improve aerodynamics after dispersal and germination takes place close to the forest floor (e.g. Evans et al. 2011a, 2011b). With this behavioral manipulation, the fungus acquires a platform to release the spores that will

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infect susceptible individuals and thus the cycle continues. In addition, the vertical

displacement of the host may also be to look for a location with an optimal

microclimate which will benefit the growth and development of the fungus and the

production of ascospores (Poulin et al. 1994) as has already been observed in the wasp-

fungus (Somavilla et al. 2019a) and ant-fungus system (Hughes et al. 2011). However,

because our study does not directly test that vertical displacement is a form of

behavioral manipulation for the benefit of spore dispersion, we think that other

alternative explanations can explain our observations. For example, we observed in the

field that ants forage and explore more in the lower parts of the vegetation, so ants

would have a harder time removing dead parasitized spiders if they were in the higher

parts of the vegetation. Another alternative explanation is that spiders infected with the

fungus may not be able to return to Drafttheir silk shelters (anonymous reviewer pers.

comm.), since all individuals of M. pacoti infected by the fungus were found outside

their natural silk shelters in the leaves. In this case, it may be that the fact of finding

infected spiders in higher places is due to the impossibility of returning to the refuge

which would benefit the dispersion of the spores by not having the barrier of the silk

refuge. However, future studies would be necessary where the location of the dead

host's body is manipulated to record the real change that height can have in the

development and fitness of the parasitic fungus. In the ant-fungus system, another

interesting aspect is that, in addition to inducing a vertical displacement in the

vegetation, the fungus induces the ants to attach through their jaws on the vegetation

(“death grip”), a behavior that is not typical of uninfected ants. Analogous to the “death

grip” in ants, in our study, dead infected spiders were found in a stretched position, a

disposition that was not registered in uninfected spiders. This position probably helps

fix the spider's postmortem on the leaves by increasing the contact surface between the

13 © The Author(s) or their Institution(s) Canadian Journal of Zoology Page 14 of 30

host and the substrate, which, consequently, will benefit the dispersion of the fungus spores. Somavilla et al. (2019b) also recorded wasps Agelaia vicina de Saussure, 1854

(Vespidae: Polistinae) killed and infected by fungi using only the legs to attach to the substrate, unlike other host wasps of the same genus, which used, in addition to the legs, the jaws to hold on to the substrate.

The population of M. pacoti infected by Gibellula sp., despite remaining relatively stable throughout the year, significantly increased with increasing rainfall.

Likewise, the rate of parasitism also had a positive correlation with the rainfall regime.

Similar results were recorded by Pontoppidan et al. (2009) where a peak in the average density of ants infected by fungi was observed during the rainy season in Thailand, which reached a density of up to 26 dead ants infected per m². Also in Thailand, another study found a significant correlationDraft between rainfall and the number of new ants killed by O. unilateralis. The record was made one month after the beginning of the rains, showing that the occurrence of this fungus in ants can follow a seasonal pattern

(Mongkolsamrit et al. 2012). In the Amazon rainforest, over a period of 14 months, there was also an increase in the abundance of ants infected by O. unilateralis at higher heights in relation to the soil during the period of the year with the highest precipitation and the highest relative humidity (Cardoso-Neto et al. 2019). Probably the high humidity in the environment benefits the development of the araneopathogenic fungus, wherein the optimum humidity and temperature conditions for the development of the fungus infection would be between 20-30 °C and above 90 % humidity respectively

(Tanada and Kaya 1993). However, we must take into account that the registered values may be underestimated, because the criterion we used to identify the parasitized spiders was the fact that they are already dead and fixed on the leaves. As a result, it is likely

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that spiders already parasitized that are not yet in the spore/hyphae stage have not been

counted as parasitized spiders.

Finally, we believe that the araneopathogenic fungus must represent an

important mortality factor for the population of M. pacoti, especially during periods of

greater precipitation. Although we did not record changes in abundance in the spider

population, it was possible to register a large amount of dead infected spiders at the

study site just during the rainy season (personal obs.). However, future studies are

needed to show the real effect that the fungus may have on the population of M. pacoti

as well as other fungus-spider interactions since they are topics that are still little

explored.

ACKNOWLEDGMENT

This work was carried out with supportDraft from the Coordenação de Aperfeiçoamento de

Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001 (I.D.

Paiva-Arruda), the Fundação Coordenação de Aperfeiçoamento de Pessoal de Nível

Superior – CAPES, process number 88887.136354/2017-00 (G.A Villanueva-Bonilla),

the Instituto Nacional de Ciência e Tecnologia dos Hymenoptera Parasitoides

(HYMPAR-CNPq/FAPESP/CAPES) and the FUNCAP– BPI proc. BP3-00139-

00186.01.00/18. We would like to thank Prof. Dr. João Paulo Machado de Araújo for

the identification of parasitic fungi species. Thanks also to Wermerson Ribeiro dos

Santos, Emily de Oliveira Fonseca, Joedson Pires, Julie Erica, Francisco Ageu Nobrega,

Brenda Kelly Souza Santiago and Lilian Araujo for field assistance, and to Mrs. Cláudia

Maria M.B. from Goes, do Sítio São Luís, Pacoti-CE, for granting access to the study

area. We are grateful to the editor and the two reviewers for their appropriate and

constructive suggestions to improve the paper.

15 © The Author(s) or their Institution(s) Canadian Journal of Zoology Page 16 of 30

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Figure captions

Figure 1. Climate diagram of Serra de Baturité, Ceará, Brazil, based on weather data collected by INMET (national meteorological institute) between 2018 and 2019. Dark areas indicate super-wet periods. Areas with vertical lines indicate humid periods.

Dotted areas indicate dry periods.

Figure 2. Location of the study area in the municipality of Pacoti, in the Serra de

Baturité Environmental Protection Area, located in the state of Ceará, Brazil. Figure was created using QGIS version 2.18.28 and assembled from the data sources

(shapefiles) of the IBGE (Instituto Brasileiro de Geografia e Estatística) and IPECE

(Instituto de Pesquisa e Estratégia Econômica do Ceará). Figure 3. Individuals of MacrophyesDraft pacoti infected with fungi Gibellula sp. A-B. Dead spiders with body covered with fungi Gibellula sp. on the abaxial surface of the leaves. C. Dead spider showing in detail the reproductive structures of the fungi that cover the host's body.

Figure 4. Silk shelter built by Macrophyes pacoti located in the abaxial part of the leaves close to the central rib. A. Female of M. pacoti. B. Female and male inside the silk shelter.

Figure 5. Comparison of the mean height in relation to the ground between infected and uninfected spiders by the araneopathogenic fungus Gibellula sp. on trees. Different letters indicate statistically significant differences.

Figure 6. Circular histograms of the abundance of M. pacoti infected by Gibellula sp.

The black vector line inside the circle indicates the average angle or the average direction of the data. The cross line outside the circle indicates the 95% confidence interval.

Figure 7. Result of the correlation coefficient between: (A) total abundance of infected

spiders and monthly precipitation (Pearson test). (B) Rate of spiders parasitism to

monthly precipitation (Spearman test). Registration made from September 2018 to

August 2019. The gray area indicates the 95% confidence interval.

Table captions

Table 1. Circular statistics applied to abundance of parasitized spiders by the fungus

Gibellula sp. Serra de Baturité, Ceará, Brazil.

Parasitized spiders Number of records 357 Mean vector ( µ ) 108,608° Month April 95% confidence interval (-/+) for ( µ ) 87.797° 129.42° Period of duration (confidence interval) 30 March Draft to 11 May Mean vector's length ( r ) 0.083 Rayleigh's test 14.513 Rayleigh's test ( p ) 4.98E-07

Draft