Synonymy of Ixodes aragaoi with Ixodes fuscipes, and reinstatement of Ixodes spinosus (Acari: Ixodidae)

Journal Pre-proof Synonymy of Ixodes aragaoi with Ixodes fuscipes, and reinstatement of Ixodes spinosus (Acari: Ixodidae) Marcelo B. Labruna (Conceptualization) (Data curation) (Formal analysis) (Investigation) (Methodology) (Resources) (Supervision) (Validation) (Visualization) (Writing - original draft) (Writing - review and editing), Valeria C. Onofrio (Conceptualization) (Data curation) (Formal analysis) (Investigation) (Methodology) (Project administration) (Resources) (Validation) (Visualization) (Writing original draft) (Writing - review and editing), Darci M. Barros-Battesti (Data curation) (Investigation) (Methodology) (Project administration) (Resources) (Supervision) (Validation) (Writing review and editing), Sergio L. Gianizella (Data curation) (Investigation) (Resources) (Validation) (Writing - review and editing), Jose´ M. Venzal (Data curation) (Investigation) (Resources) (Validation) (Writing - review and editing), Alberto A. Guglielmone (Conceptualization) (Formal analysis) (Investigation) (Validation) (Visualization) (Writing - original draft) (Writing - review and editing)

PII:

S1877-959X(19)30361-9

DOI:

https://doi.org/10.1016/j.ttbdis.2019.101349

Reference:

TTBDIS 101349

To appear in:

Ticks and Tick-borne Diseases

Received Date:

31 August 2019

Revised Date:

19 November 2019

Accepted Date:

28 November 2019

Please cite this article as: Labruna MB, Onofrio VC, Barros-Battesti DM, Gianizella SL, Venzal JM, Guglielmone AA, Synonymy of Ixodes aragaoi with Ixodes fuscipes, and reinstatement of Ixodes spinosus (Acari: Ixodidae), Ticks and Tick-borne Diseases (2019),

doi: https://doi.org/10.1016/j.ttbdis.2019.101349

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Synonymy of Ixodes aragaoi with Ixodes fuscipes, and reinstatement of Ixodes spinosus (Acari: Ixodidae)

Marcelo B. Labrunaa*, Valeria C. Onofriob,c, Darci M. Barros-Battestid, Sergio L. Gianizellae, José M. Venzalf, Alberto A. Guglielmoneg

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Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São

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Paulo, SP, Brazil Mestrado em Medicina e Bem Estar Animal, Universidade Santo Amaro, São Paulo,

SP, Brazil

Laboratório Especial de Coleções Zoológicas, Instituto Butantan, São Paulo, SP,

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c

Brazil

Departamento de Patologia Veterinária, Faculdade de Ciências Agrárias e

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Veterinárias, Universidade Estadual Paulista - UNESP, Jaboticabal, SP, Brazil. Laboratório de Zoologia, Departamento de Biologia, Instituto de Ciências Biológicas,

Universidade Federal do Amazonas, Manaus, AM, Brazil. Laboratorio de Vectores y enfermedades transmitidas, Facultad de Veterinaria,

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CENUR Litoral Norte, Universidad de la República, Salto, Uruguay. Instituto Nacional de Tecnología Agropecuaria, Estación Experimental Agropecuaria

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Rafaela, Consejo Nacional de Investigaciones Científicas y Técnicas, Rafaela, Santa Fe, Argentina.

* Corresponding author. Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP, Brazil, 05508-270

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Tel.: +55 11 3091-1394 E-mail address: [email protected] (M.B. Labruna)

Abstract. Ixodes fuscipes Koch, 1844, described from Brazil, has been considered a valid species with records from Brazil and Peru. Ixodes spinosus Neumann, 1899, also described

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from Brazil, has been considered a synonym of I. fuscipes. In 2014, Ixodes aragaoi Fonseca 1935 was redescribed as a valid species for Brazil and Uruguay. Until the

present study, one major difference between the females of I. fuscipes and I. aragaoi

was the horn-like auriculae in the former versus the ridge-like auriculae in the later, but

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this morphological difference was not supported after examination of the holotype of I.

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fuscipes and a syntype female of I. spinosus. Surprisingly, we found the type of I. fuscipes to have ridge-like auriculae, in contrast to the horn-like auriculae of I. spinosus.

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Comparisons of the I. fuscipes holotype with the syntypes of I. aragaoi revealed that they correspond to the same species. Therefore, we redescribe I. fuscipes, relegate I.

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aragaoi to a junior synonym of I. fuscipes, and reinstate I. spinosus as a valid species. After examining all lots of I. fuscipes in four tick collections, no specimen was

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recognized as I. fuscipes when compared with the type specimen of this taxon. On the other hand, specimens previously identified as I. aragaoi are now confirmed as I.

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fuscipes, with bona fide records for Brazil and Uruguay. Some of the specimens previously reported as I. fuscipes are now confirmed as I. spinosus, with records in two Brazilian biomes, Amazon and Atlantic rainforest. We present lists of the ticks examined in this study, with their current taxonomic status.

Keywords: Ixodids; ticks; taxonomy; South America 2

1. Introduction

Koch (1844) described the species Ixodes fuscipes Koch, 1844 based on one female collected in Brazil. The morphological description provided by Koch (1844) is superficial, consisting of only three text lines without illustrations, with no host or locality data in addition to Brazil. Three years later, Koch (1847) redescribed I. fuscipes,

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still superficially, providing a rough drawing of the dorsal view of the specimen. Neumann (1899) described the species Ixodes spinosus Neumann, 1899 based on four females, seven nymphs and three larvae collected by E. Goeldi on Dasyprocta

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aguti in Brazil, with no precise locality data or illustrations. After examining the type of I. fuscipes, Neumann (1901) synonymized I. spinosus with I. fuscipes. Since then, this

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synonymy has been accepted (Guglielmone and Nava, 2014).

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Nuttall and Warburton (1911) admitted the synonymy proposed by Neumann (1901), and redescribed I. fuscipes based on one female from the type series of I. spinosus. In this case, Nuttall and Warburton (1911) provided a detailed morphological

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description and figures of the female, showing several key characters (i.e., horn-like auriculae, ventral spurs on palpal article I, coxa I with a long internal spur and a short

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external spur, coxae II-IV each with a short external spur) that have been used until

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nowadays for morphological identification of I. fuscipes in South America. Ixodes fuscipes has been reported from Peru (Fairchild et al., 1966; Need et al.,

1991) and Brazil (Koch, 1844; Neumann, 1899; Rohr, 1909; Aragão, 1911; 1936; Nuttall and Warburton, 1911; Barros-Battesti and Knysak, 1999; Arzua et al., 2005; Labruna et al., 2005a; Martins et al., 2014; Blanco et al., 2017; Michel et al., 2017; Gianizella et al., 2018). Although I. fuscipes was recorded in Panama (Nuttall and

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Warburton, 1911), this record could not be confirmed (Fairchild et al., 1966; Guglielmone et al., 2003). In addition, a record of an I. fuscipes female from Colombia (Neumann, 1911) seems to refer to the same female specimen reported by Nuttall and Warburton (1911) for Panama, as both records coincide in the host species (Felis pardalis), and Panama was politically part of Colombia until 1903. The validity of the taxon Ixodes aragaoi Fonseca 1935 was a subject of much controversy until the study of Onofrio et al. (2014), who validated the species and

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redescribed it based on examinations of the type specimens and material from several other tick collections. The geographical distribution of I. aragaoi has been restricted to

Brazil and Uruguay (Onofrio et al., 2014; Nava et al., 2017). According to Onofrio et al.

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(2014), I. aragaoi belongs to the I. ricinus species complex, whose females are

characterized by auriculae as indistinct lateral extensions (=ridge-like auriculae).

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Therefore, until the present study, one major difference between the females of I.

auriculae in the later.

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fuscipes and I. aragaoi was the horn-like auriculae in the former versus the ridge-like

For the present study, we examined the holotype specimen of I. fuscipes and one

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female of the syntype series of I. spinosus, which is the only specimen left from the original series composed by four females, seven nymphs and three larvae. Surprisingly,

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we found the type of I. fuscipes to have ridge-like auriculae, in contrast to the horn-like

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auriculae of I. spinosus. Comparisons of the I. fuscipes holotype with the syntypes of I. aragaoi revealed that they correspond to the same species. Therefore, we redescribe I. fuscipes, relegate I. aragaoi to a junior synonym of I. fuscipes, and reinstate I. spinosus as a valid species.

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2. Materials and methods

2.1. List of revised collections and their acronyms Zoologisches Museum Berlin, Berlin, Germany (ZMB); École Nationale Vétérinaire, Toulouse, France (ENV); Coleção Acarológica do Instituto Butantan, São Paulo, SP, Brazil (IBSP); Coleção Nacional de Carrapatos “Danilo Gonçalves Saraiva”, São Paulo, SP, Brazil (CNC); Coleção Zoológica Paulo Bürhnheim da Universidade Federal do

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Amazonas, Manaus, AM, Brazil (CZPB); United States National Tick Collection, Statesboro, GA, United States (USNMENT); Departmento de Parasitología Veterinaria,

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Facultad de Veterinaria, UdelaR, Salto, Uruguay (DPVURU).

2.2. Type material examined

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Ixodes fuscipes - ZMB 1123, holotype, 1 female, Brazil, host and collection date

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unknown.

Ixodes spinosus - ENV 750, syntype, 1 female, ex Dasyprocta aguti, Brazil, 1898. Ixodes aragaoi - IBSP 744, syntypes, 4 males, 5 females, ex Mazama gouazoubira, São

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Paulo municipality, São Paulo state, Brazil, 21/V/1934; IBSP 829, syntypes, 8 males, 21 females, ex Mazama gouazoubira, Cotia muncipality, São Paulo state, Brazil,

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05/X/1934.

2.3. Other material examined We examined all lots of I. fuscipes and I. aragaoi in the tick collections IBSP, CNC, CZPB, and DPVURU. These included 96 males, 180 females from IBSP, 11 males, 27 females from CNC, 143 females from CZPB, and 63 males, 217 females from

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DPVURU. In addition, the only two lots of I. fuscipes (2 females) in the USNMENT were also examined.

2.4. Material preparation Light photographs of the type specimens of I. fuscipes and I. spinosus were taken at the ZMB and ENV tick collections, respectively, with the aid of the curator of each of these collections. Selected specimens deposited in Brazilian collections were cleaned for both

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light and scanning electron microscopy according to Guimarães et al. (2001). Measurements were taken under a Leica M205A stereomicroscope. Measurements are given in micrometers, with the mean followed by the standard deviation and range in

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parentheses. Light microscopy photographs were taken under a stereomicroscope (Zeiss Stemi SV 11, Zeiss, Munich, Germany) using the software ZEN 2 core. The images

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were prepared with Adobe Photoshop v. 6, and Corel DRAW X5, v. 12.

3. Results

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3.1. Redescription of the type specimen of Ixodes fuscipes Koch, 1844 ZMB 1123, holotype, 1 female, Brazil, host and collection date unknown.

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Type locality: Brazil. While Koch (1844) did not provide the locality name in the

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published description, the type label indicated “Casapav, Brazil”, which could refer to Caçapava Municipality, São Paulo state (southeastern Brazil), or less probably to Caçapava do Sul, Rio Grande do Sul state (southern Brazil). Due to the critical and fragile condition of the holotype of I. fuscipes, it could not be measured for the present redescription.

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Female (Fig. 1A-D) Gnathosoma: Dorsal basis capituli subtriangular, posterior margin slightly concave; porose areas transversally elongated, separated by distance shorter than their own width; cornua absent (Fig. 1A). Ventral basis capituli narrowed posterior to auriculae, with straight posterior margin and lateral angles rounded; apparent transverse suture; ridgelike auriculae; palpal article I without ventral spur (Fig. 1B,C). Hypostome elongated, narrowly rounded at apex; dental formula 4/4 for most of length, then 3/3 and 2/2 near

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base, corona with numerous denticles. Idiosoma: Scutum rounded with large punctations, scapulae prominent, but posterior scutal field partly missed due to pinned holotype specimen. Ventral surface with

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numerous and long setae, spiracular plate oval.

Legs: Coxae I with a straight and long internal spur, with sharply pointed apex, reaching

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the mid-length of coxa II, with a short and slightly triangular external spur; coxae I-II

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each without syncoxal areas; coxae II-IV each with a short and slightly triangular

3.1.1. Remarks

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external spur, without internal spur (Fig. 1D).

After examining all lots of I. fuscipes in the IBSP, CNC, CZPB, USNMENT and

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DPVURU tick collections, no specimen was confirmed as I. fuscipes when compared

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with the type specimen of this taxon (ZMB 1123). Examination of a type female of I. spinosus revealed that it differs from the I. fuscipes type by having long, pointed, hornlike auriculae, suboval porose areas, cornua with rounded apices, and palpal article I with a well-developed ventral spur, elongated and tapering apically. Therefore, I. spinosus Neumann, 1899 is reinstated as a valid species (redescription below).

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Examination of the type females of I. aragaoi revealed that they present the same morphological traits of I. fuscipes (Fig. 2A-F) and in the recent redescription of I. aragaoi reported by Onofrio et al. (2014). Therefore, I. aragaoi Fonseca, 1935 is relegated to a junior synonym of I. fuscipes. Because of the synonymia, the I. aragaoi male described in Onofrio et al. (2014) serves now as I. fuscipes male description. Thus, at present, there are descriptions of both sexes. Table 1 lists the specimens examined and confirmed as I. fuscipes in the present

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study. Except for the holotype female of I. fuscipes, the latest reports of all remaining specimens in Table 1 referred to I. aragaoi, which is now a valid synonym of I.

fuscipes. Based on these confirmed records, the geographical distribution of I. fuscipes

Paraná and Rio Grande do Sul), and Uruguay.

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has been restricted to southeastern Brazil (state of São Paulo), southern Brazil (states of

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During our examinations of the tick collections, only two lots of I. aragaoi were

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not confirmed as I. fuscipes. One lot (IBSP4457), published as I. aragaoi by BarrosBattesti and Knysak (1999), refers to one female collected on a passerine bird in São Paulo municipality. Our examination of this female tick revealed its identification as

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Ixodes auritulus Neumann, 1904. Another lot (CNC617), published as I. aragaoi by Labruna et al. (2005b), contains 1 male and 1 female that were collected on Puma

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concolor in Barão de Melgaço, state of Mato Grosso, near to the border of Brazil with

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Bolivia. They differ from I. fuscipes regarding the shape of the three transversal lobes in male ventral capituli, and the punctuation pattern in the female scutum. Therefore, we are relegating this lot to Ixodes sp. until further studies define its taxonomic status. Artiodactyla mammals (chiefly the native deer M. gouazoubira) are major hosts for the adult stage of I. fuscipes (Table 1). In Uruguay, cattle are also important hosts (Nava et al., 2017). As for the immatures, there have been various records of I. fuscipes

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(all reported as I. aragaoi or Ixodes pararicinus Keirans & Clifford, 1985) larvae and nymphs infesting small mammals and birds in Brazil and Uruguay (Venzal et al., 2005; Arzua et al., 2005; Onofrio et al., 2014; Blanco et al., 2017; Michel et al., 2017; Nava et al., 2017).

3.2. Redescription of a type female of Ixodes spinosus Neumann, 1899 ENV 750, syntype, 1 female, ex. Dasyprocta aguti, Brazil, col. E. Goeldi, 1898. We

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designated this specimen as the lectotype of I. spinosus. Neumann (1899) described I. spinosus based on 4 females, 7 nymphs and 3 larvae; however, only the female designated here as lectotype for this species remains in the ENV collection.

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Type locality: Brazil [no more information available for type locality in Neumann (1899)]

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During our examination of the type specimen at the ENV collection, we were not

Female (Fig. 3A-E)

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allowed to take measurements of the specimen, due to its fragile condition.

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Gnathosoma: Dorsal basis capituli subtriangular, posterior margin nearly straight; suboval porose areas separated by distance approximately equal to their own width;

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cornua moderated with rounded apices (Fig. 3A, D). Ventral basis capituli narrowed

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posterior to auriculae, with straight posterior margin and lateral angles rounded; apparent transverse suture; long, pointed, horn-like auriculae (Fig. 3B, C, E). Palpi elongate and slender, article I with a well-developed ventral spur, elongated and tapering apically. Hypostome elongated, narrowly rounded at apex; dental formula 3/3 for most of length, then 2/2 near base, corona with numerous denticles (Fig. 3B)

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Idiosoma: broadly suboval, widest slightly posterior to mid-length. Scutum with larger punctations clustered posteriorly and in antero-lateral fields, few fine punctations in the central area; cervical grooves distinct, moderately shallow and reaching the posterior third of the scutum; lateral carinae as moderately sharp ridges; scapulae prominent (Fig. 3D). Ventral surface with numerous and long setae evenly distributed; genital aperture medial to coxae IV (Fig. 3E). Legs: moderately long, slender. Coxae I with a straight and long internal spur, with

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sharply pointed apex, not reaching the mid-length of coxa II, with a short and slightly triangular external spur; coxae I-II each without syncoxal areas; coxae II-IV each with a

3.3. Additional specimens of I. spinosus

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short and slightly triangular external spur, without internal spur (Fig. 3C, E).

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After examining all lots of I. fuscipes and I. aragaoi in the tick collections IBSP, CNC,

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CZPB, USNMENT and DPVURU, we were able to identify ten female specimens as I. spinosus (previously identified as I. fuscipes, Table 2). One of these females was subsequently deposited at the USNMENT collection under the number

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USNMENT860091 (Table 2). Seven of these females have been previously reported as I. fuscipes, as shown in Table 2. Four of these females were used for a more detailed

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redescription, as stated below.

3.3.1. Redescription of Ixodes spinosus Neumann, 1899 based on female specimens collected in Rondônia state, Brazil Brazil - Rondônia state: Cacaulândia municipality (10º18’S, 63º14’W): 1 ♀, collected 09/X/2001 on vegetation, deposited at the CNC 571 [this female was previously illustrated by Onofrio et al. (2009), as I. fuscipes]; Monte Negro municipality (10º15’S,

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63º19’W): 1 ♀, collected 07/XII/2001 on Dasyprocta sp., deposited at the CNC 572; 1 ♀, collected 27/I/2005 on Dasyprocta sp., deposited at the CNC 887; 1 ♀, collected 27/I/2005 on Dasyprocta sp., deposited at the USNMENT 00860091. Redescription based on one unfed (CNC 571) and three partially engorged females (CNC 572, 887; USNMENT 00860091). Only one female had intact tarsus I; all had broken tarsus IV.

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Female (Fig. 4A-D) Gnathosoma: length from palpal apices to posterior margin of basis capituli 1,259 ± 81 (1,182 - 1,343), width of basis capituli dorsally 571 ± 8 (562 - 582), ratio 2.23 ± 0.11

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(2.13 - 2.36). Dorsal basis capituli subtriangular, posterior margin nearly straight,

length from medial insertion of palpal segment I to cornual apices 275 ± 6 (267 - 281),

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ratio width to length 2.08 ± 0.05 (2.02 - 2.14); suboval and small porose areas indented

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with well-circumscribed borders, separated by distance approximately equal to their own width; cornua moderated with rounded apices (Fig. 4A). Ventral basis capituli narrowed posterior to auriculae, with straight posterior margin and lateral angles

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rounded; apparent transverse suture; long, pointed, horn-like auriculae, length from basis to tips 176 ± 12 (165 - 188), width of basis 83 ± 8 (73 - 93), ratio 2.14 ± 0.11 (2.02

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- 2.26) (Fig. 4B). Palpi elongate and slender, total length 1,004 ± 45 (975 - 1,056),

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width 203 ± 12 (191 - 215), ratio 4.94 ± 0.18 (4.78-5.14), length of segments in descending order: 2, 3, 1, 4; basis of article I with a well-developed ventral spur, elongated and tapering apically, length 174 ± 13 (158 - 189), width 73 ± 4 (67 - 75), ratio 2,40 ± 0.14 (2.27 - 2.59). Hypostome length 717 ± 66 (670 - 763), width 162 ± 8 (156 - 167), ratio 4.43 ± 0.20 (4.29 - 4.57), elongated, narrowly rounded at apex; dental formula 3/3 for most of the length, then 2/2 near base, corona with numerous denticles.

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Idiosoma: broadly suboval, widest slightly posterior to mid-length. Dorsal surface length from scapular apices to posterior body margin (one unfed and three partially engorged specimens) 3,459 ± 927 (2,125 - 4,249); maximum width 2,360 ± 623 (1,439 2,811), ratio 1.47 ± 0.1 (1.37 - 1.61). Scutum length 1,578 ± 75 (1,512 -1,684), width 1,408 ± 76 (1,324 - 1,487), ratio 1.12 ± 0.05 (1.05 - 1.16), elongated; larger punctations clustered posteriorly and in antero-lateral fields, few fine punctations in the central area; cervical grooves distinct, moderately shallow and reaching the posterior third of the

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scutum; lateral carinae as moderately sharp ridges; scapulae prominent (Fig. 4C). Ventral surface with numerous and long setae evenly distributed; genital aperture medial to coxae IV.

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Legs: moderately long, slender. Coxae I length 348 ± 24 (317 - 369), maximum width 306 ± 8 (300 - 318), ratio 1.14 ± 0.06 (1.06 - 1.21), with a straight and long internal

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spur, with sharply pointed apex, not reaching the mid-length of coxa II, length from

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posterior seta of coxa I to the tip of the spur 166 ± 20 (139 - 185), width of basis 66 ± 4 (62 - 72), ratio 2.52 ± 0.36 (2.14 - 2.85), with a short and slightly triangular external spur; coxae I-II each without syncoxal areas; coxae II-IV each with a short and slightly

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triangular external spur, without internal spur (Fig. 4D). Tarsus I length 834, width 204,

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ratio 4.09; tarsus IV broken in all measured specimens.

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3.4. Remarks

While the Brazilian locality of the type series of I. spinosus was not detailed by

Neumann (1899), the additional specimens reported in the present study confirms I. spinosus in at least two biomes, Amazon (Peru and the Brazilian states of Rondônia and Acre) and Atlantic rainforest (Brazilian state of Rio de Janeiro). All host records of I.

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spinosus female were on Cavioidea rodents of the genera Dasyprocta, Myoprocta and Cuniculus (Table 2). Immature stages of I. spinosus were unavailable for this study but Neumann (1899) listed Dasyprocta aguti as host for the larva and nymph of this species. In Table 3, we list adult ticks that were previously reported as I. fuscipes, but after morphological analyses in the present study, they were shown to be whose identification need further exploration as they are distinct from either both I. fuscipes or

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and I. spinosus. The taxonomic status of these specimens will be presented in a separate report (in preparation).

In Table 4 we list previous reports of I. fuscipes that could not be reevaluated in

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the present study. Due to the uncertainty of the taxonomic status of these reports, each

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of them should be relegated to Ixodes sp. until further analyses.

AUTHOR STATEMENT

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M. B. Labruna, V. C. Onofrio, D. M. Barros-Battesti, S. L. Gianizella, J. M. Venzal, A.

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A. Guglielmone

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MBL, VCO, AAG: Conceptualization; MBL, VCO, DMB-B, SLG, JMV: Data curation; MBL, VCO, AAG: Formal analysis; MBL, VCO, DMB-B, SLG, JMV, AAG: Investigation; MBL, VCO, DMB-B: Methodology; VCO, DMB-B: Project administration; MBL, VCO, DMB-B, SLG, JMZ: Resources; MBL, DMB-B: Supervision; MBL, VCO, DMB-B, SLG, JMV, AAG: Validation; MBL, VCO, AAG: Visualization; Roles/Writing - original draft; MBL, VCO, DMB-B, SLG, JMV, AAG: Writing - review & editing.

Acknowledgements

We are grateful to Jason Dunlop (curator of the ZMB) and Michel Franc (curator of the ENV) for allowing examination and imaging of the type specimens of I. fuscipes and I. 13

spinosus. We are also thankful to Dmitry A. Apanaskevich for providing details on specimens of I. fuscipes stored in the USNMENT.

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Guglielmone, A.A., Estrada-Peña, A., Keirans, J.E., Robbins, R.G., 2003. Ticks (Acari:

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Veterinária. Ed. Plêiade/FAPESP, São Paulo, 218 pp.

Koch, C.L., 1844. Systematische Übersicht über die Ordnung der Zecken. Arch. Naturgesch. 10, 217-239.

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Koch, C.L., 1847. Übersicht des Arachnidensystems. Nürnberg, 4, 136 pp. Labruna, M.B., Camargo, L.M.A., Terrassini, F.A., Ferreira, F., Schumaker, T.T.,

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Camargo, E.P., 2005a. Ticks (Acari: Ixodidae) from the state of Rondônia, western

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Amazon, Brazil. Syst. Appl. Acarol. 10, 17–32. Labruna, M.B., Jorge, R.S., Sana, D.A., Jácomo, A.T., Kashivakura, C.K., Furtado, M.M., Ferro, C., Perez, S.A., Silveira, L., Santos, T.S. Jr, Marques, S.R., Morato, R.G., Nava, A., Adania, C.H., Teixeira, R.H., Gomes, A.A., Conforti, V.A., Azevedo, F.C., Prada, C.S., Silva, J.C., Batista, A.F., Marvulo, M.F., Morato, R.L.,

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Alho, C.J., Pinter, A., Ferreira, P.M., Ferreira, F., Barros-Battesti, D.M., 2005b. Ticks (Acari: Ixodida) on wild carnivores in Brazil. Exp. Appl. Acarol. 36, 149-163. Martins, T.F., Venzal, J.M., Terassini, F.A., Costa, F.B., Marcili, A., Camargo, L.M., Barros-Battesti, D.M., Labruna, M.B., 2014. New tick records from the state of Rondônia, western Amazon, Brazil. Exp. Appl. Acarol. 62, 121-128. Martins, T.F., Teixeira, R.H.F., Labruna, M.B., 2015. Occurrence of ticks on wild animals received and attended at the Parque Zoológico Municipal Quinzinho de

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Barros, Sorocaba, São Paulo, Brazil. Braz. J. Vet. Res. Anim. Sci., São Paulo 52, 319-324.

Michel, T., Souza, U., Dall’Agnol, B., Webster, A., Peters, F., Christoff, A., Luza, A.L.,

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M.M.A., Ott, R., Reck, J., 2017. Ixodes spp. (Acari: Ixodida) ticks in Rio Grande do

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Sul State, Brazil. Syst. Appl. Acarol. 22, 2057-2067.

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Nava, S., Venzal, J.M., González-Acuña, D., Martins, T.F., Guglielmone, A.A., 2017. Ticks of the Southern Cone of America. Elsevier Academic Press, London, San Diego, Cambridge, Oxford, 348 pp.

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Need, J.T., Dale, W.E., Keirans, J.E., Dasch, G.A., 1991. Annotated list of ticks (Acari: Ixodidae, Argasidae) reported in Peru: distribution, hosts, and bibliography. J. Med.

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Entomol. 28, 590-597.

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Neumann, L.G., 1899. Révision de la famille des Ixodidés (3e mémoire). Mém. Soc. Zool. Fr. 12, 107-294.

Neumann, L.G., 1901. Révisión de la famille des ixodidés (4e mémoire). Mém. Soc. Zool. Fr. 14, 249-372. Neumann, LG. 1911. Ixodidae. Das Tierreich, 26, 169 pp.

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Nuttall, G.H.F., Warburton, C., 1911. Ticks. A monograph of the Ixodoidea. Part II. Ixodidae. Cambridge Univ. Press, London, pp. 105–348. Onofrio, V.C., Barros-Battesti, D.M., Labruna, M.B., Faccini, J.L.H. 2009. Diagnoses of and illustrated key to the species of Ixodes Latreille, 1795 (Acari: Ixodidae) from Brazil. Syst. Parasitol. 72, 143-157. Onofrio, V.C., Ramírez, D.G., Giovanni, D.N.S., Marcili, A., Mangold, A.J., Venzal, J.M., Mendonça, R.Z., Labrunam M.B., Barros-Battesti, D.M., 2014. Validation of

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the taxon Ixodes aragaoi Fonseca (Acari: Ixodidae) based on morphological and molecular data. Zootaxa, 3860, 361-370.

Rohr, C., 1909. Estudo sobre Ixódidas do Brasil. Inst. Oswaldo Cruz, Rio de Janeiro,

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Sabatini, G.S., Pinter, A., Nieri-Bastos, F.A., Marcili, A., Labruna, M.B., 2010. Survey

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of ticks (Acari: Ixodidae) and their rickettsia in an Atlantic rain forest reserve in the

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State of São Paulo, Brazil. J. Med. Entomol. 47, 913-916. Saracho-Bottero, M.N., Venzal, J.M., Tarragona, E.L., Thompson, C.S., Mangold, A.J., Beati, L., Guglielmone, A.A., Nava, S., 2019. Ixodes ricinus complex (Acari:

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Ixodidae) in the Southern Cone of America: Ixodes pararicinus, Ixodes aragaoi and Ixodes sp. cf. I. affinis. Parasitol. Res. (In press).

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Teixeira, R.H.F., Labruna, M.B., Martins, T.F., 2017. Hard ticks collected parasitizing

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wild animals at Sorocaba Zoo, state of São Paulo, Brazil. Revista de Educação Continuada em Medicina Veterinária e Zootecnia do CRMV-SP 15, 8-14.

Venzal, J.M., Estrada-Peña, A., Barros-Battesti, D.M., Onofrio, V.C., Beldomenico, P.M., 2005. Ixodes (Ixodes) pararicinus Keirans & Clifford, 1985 (Acari: Ixodidae): description of the immature stages, distribution, hosts and medical/veterinary importance. Syst. Parasitol. 60, 225–234.

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Figure legends

Fig. 1. Holotype female of Ixodes fuscipes. A. Dorsal basis capituli (bar: 200 µm). B. Ventral basis capituli and coxae I-IV (bar: 500 µm). C. Ventral basis capituli (bar: 200 µm). D. Ventral view of the specimen, with missing legs and palpi, and broken idiosoma (bar: 1000 µm). E. Dorsal view; damaged idiosoma due to pinned conservation (bar: 1000 µm). F. Ventral view; damaged idiosoma due to pinned

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conservation (bar: 1000 µm).

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Fig. 2. Type females of Ixodes aragaoi (IBSP 829). A. Ventral capitulum (bar: 200 µm). B. Ventral basis capituli (bar: 100 µm). C. Dorsal basis capituli and scutum (bar:

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200 µm). D. Coxae I-IV (bar: 200 µm).

Fig. 3. Lectotype female of Ixodes spinosus. A. Dorsal view of the specimen. B. Ventral capitulum. C. Coxae I-IV. D. Dorsal capitulum and scutum. E. Ventral capitulum and coxae I-IV.

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Fig. 4. Scanning electron microscopy of an Ixodes spinosus female (CNC 571) from

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Rondônia state, Brazil. A. Dorsal capitulum. B. Ventral capitulum. C. Scutum. D. Coxae I-IV. Bars: 200 µm. Photographs A, B, and C have been published as I. fuscipes

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by Onofrio et al. (2009); Syst. Parasitol. 72, 143–157.

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Table

Table 1. Tick specimens examined in the present study and confirmed as Ixodes fuscipes Country

State1 or Department: municipality

Source

No. specimens2

Reference

Taxonomic notes

Brazil

?: Casapav3 SP: São Paulo

? Mazama gouazoubira

1F 4M,5F

SP: São Paulo SP: São Paulo SP: São Paulo SP: São Paulo SP: São Paulo SP: Cotia

Mazama sp ? M. gouazoubira M. gouazoubira M. gouazoubira M. gouazoubira

5M,17F 1M 1F 1F 5M,6F 8M,21F

Holotype of I. fuscipes Syntypes of Ixodes aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi Syntypes of I. aragaoi

SP: Cotia SP: Cotia SP: Cotia SP: Cotia SP: Cotia SP: Cotia SP: Embu SP: Embu SP: Embu-Guaçu SP: Itap. da Serra SP: Itap. da Serra SP: Itap. da Serra SP: Lins SP: Biritiba Mirim SP: Mairiporã SP: Santo André SP: Ibiúna SP: São Roque SP: Sorocaba PR: Foz do Iguaçu RS: Pin. Machado Rocha: Laguna Negra Rocha: Laguna Negra Rocha: Laguna Negra Rocha: Laguna Negra Ag. Blancas: Lavalleja Rocha: Laguna Negra Rocha: Laguna Negra Mald.: Cerro Hermanos Mald.: Ce. Pan de Azúcar Mald.: Sierra de Valdivia Rocha: Laguna Negra Rocha: Laguna Negra Rocha: Puntas Chafalote Rocha: Laguna Negra Rocha: Laguna Negra

Mazama sp. Mazama sp. Mazama sp. Vegetation Vegetation M. gouazoubira Mazama sp. Mazama sp. Mazama sp. Mazama sp. Dog Mazama sp. Horse Molted from nymph M. gouazoubira Vegetation M. gouazoubira M. gouazoubira M. gouazoubira Mazama rufina M. gouazoubira Bos taurus Molted from nymphs B. taurus B. taurus B. taurus B. taurus B. taurus M. gouazoubira M. gouazoubira

5M,17F 7M,8F 3M,2F 2F 2M,1F 3M,2F 30M,55F 4M,7F 1M 1M 1F 12M,15F 1M 1F 2F 1F 1F 1M,2F 2M,2F 1M 1F 5F 8M,1F 2M,2F 4M,15F 1F 3M,23F 2M,3F 1M,5F 1M,6F

Koch (1844) Fonseca (1935), Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Fonseca (1935), Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Martins et al. (2015) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Sabatini et al. (2010) Teixeira et al. (2017) Teixeira et al. (2017) Martins et al. (2015) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) Onofrio et al. (2014) unpublished Saracho-Bottero et al. (2019) Saracho-Bottero et al. (2019) unpublished Saracho-Bottero et al. (2019) Saracho-Bottero et al. (2019)

B. taurus B. taurus B. taurus B. taurus B. taurus B. taurus

3F 20M,58F 3M,40F 6F 23M,40F 6M,17F 169M, 397F

Saracho-Bottero et al. (2019) unpublished unpublished Saracho-Bottero et al. (2019) unpublished unpublished

reported as I. aragaoi

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TOTAL

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Uruguay

reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi reported as I. aragaoi

Collection accession number ZMB 1123 IBSP744 IBSP1730a IBSP1778 IBSP6964c IBSP7449b CNC711 IBSP829 IBSP1730a IBSP5678 IBSP10904 IBSP11103 IBSP11114 CNC3093 IBSP5679a IBSP5778 IBSP5791 IBSP3846 IBSP5632 IBSP5701 IBSP5700 CNC728 CNC732 CNC1151 CNC3135 CNC3171 CNC283 IBSP6791a IBSP890 IBSP7956 IBSP8750 IBSP8760 DPVURU287 DPVURU289 DPVURU291 DPVURU293 DPVURU294 DPVURU295 DPVURU296 DPVURU300 DPVURU302 DPVURU313 DPVURU316 DPVURU420

reported as I. aragaoi

?: uncertain or unknown 1 Abbreviations of Brazilian states - SP: São Paulo; PR: Paraná; RS: Rio Grande do Sul 2 F: female; M: male 3 Koch (1844) did not provide the locality name in the published description, the type label indicated “Casapav, Brazil”, which could refer to Caçapava Municipality, SP, or less probably to Caçapava do Sul, RS.

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Table 2. Female specimens from Brazil and Peru, examined in the present study and confirmed as Ixodes spinosus

Brazil

Peru

State1 or province: municipality or district ?: ?

Source

No.

Reference

Taxonomic notes

Collection accession number

Dasyprocta aguti

1

Neumann (1899)

Lectotype of I. spinosus

ENV 750

AC: ?

Myoprocta pratti

2

Unpublished

RJ: Angra dos Reis

Cuniculus paca

1

reported as I. fuscipes

RJ: ?

?

1

Aragão (1936), BarrosBattesti and Knysak (1999) Unpublished

RO: Cacaulândia RO: Monte Negro RO: Monte Negro RO: Monte Negro RO: Monte Negro

Vegetation Dasyprocta sp. Dasyprocta sp. Dasyprocta sp. Dasyprocta sp.

1 1 1 1 1

Labruna et al. (2005a) Labruna et al. (2005a) Labruna et al. (2005a) Labruna et al. (2005a) Labruna et al. (2005a)

reported as I. fuscipes reported as I. fuscipes reported as I. fuscipes reported as I. fuscipes reported as I. fuscipes

Sandia (Puno): San Juan del Oro

Agouti

1

Fairchild et al. (1966), Need et al. (1991)

reported as I. fuscipes

IBSP741 USNMENT98840 2 CNC571 CNC572 CNC382 CNC887 USNMENT86009 1 USNMENT98837 8

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TOTAL 11 1 Abbreviations of Brazilian states – AC: Acre ; RJ: Rio de Janeiro; RO: Rondônia ?: uncertain or unknown

IBSP7299

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Country

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Table 3. Brazilian tick specimens examined in the present study, shown to be morphologically distinct from either Ixodes fuscipes or Ixodes spinosus. State 1: municipality AM: Manaus

Source

AM: Manaus

No. specimens2 1M, 72F

Previous classification Ixodes c.f. fuscipes

Reference

D. leporina, Dasyprocta sp, Tamandua tetradactyla

84F

I. fuscipes

Unpublished

RO: Pimenta Bueno

Dasyprocta variegata

2F

I. fuscipes

SC: Capão Alto SP: Piedade

Sooretamys angouya Marmosa demerarae

1F 2F

I. fuscipes I. fuscipes

Martins et al. (2014), Onofrio et al. (2014) Unpublished Unpublished

SP: Caieiras

Felis catus

2F

I. fuscipes

Unpublished

Dasyprocta 25eporine, Dasyprocta agouti, Dasyprocta sp, Bradypus tridactylus, Cyclopes didactylus

Gianizella et al (2018)

Collection accession number CNC2488, CZPB91, 3173, 347, 384, IBSP-5736, 5737, 12721, 12722, 12723 CZPB- 336, 770, 793, 803, 839, 840, 900, 904, 905 CNC1068, IBSP1599 IBSP11981 IBSP9337, USNMENT860090 IBSP13200, CNC4030

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TOTAL 1M, 163F 1 Abbreviations of Brazilian states – AM: Amazonas; RO: Rondônia; SC: Santa Catarina, SP: São Paulo; 2 F: female; M: male 3 Gianizella et al. (2018) published lot CZPB317 as for Presidente Figueiredo municipality, but the correct municipality is Manaus.

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Table 4. Previously reports of Ixodes fuscipes that should be relegated to Ixodes sp. (undetermined species) until further analyses. Country Panama Brazil

Locality1 ? AM: Manaus

Source Felis pardalis Tamandua tetradactyla, Dasyprocta leporina, Dasyprocta sp. D. agouti or Cuniculus paca D. agouti or Cuniculus paca ? Birds D. agouti or Cuniculus paca Dasyprocta sp., small rodent Akodon paranaensis Lutreolina crassicaudata

stage F N,L

Reference Nuttall and Warburton (1911) Gianizella et al 2018

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MG: Decoberto F Rohr (1909) MG: Pacau F Aragão (1936) PA: ? F Aragão (1936) PR: Adrianópolis, Pirarquara N,L Arzua et al. (2005) RJ: Macabu F Rohr (1909) RO: Monte Negro N Labruna et al. (2005)a RS: Sao Francisco de Paula N Michel et al. (2017) SC: Floriaópolis N Arzua et al. (2005) SC: Jaborá F Blanco et al. (2017) Oligoryzomys nigripes SC: Ponte Alta do Norte Wild rodent L Arzua et al. (2005) SP: São Paulo Small rodents N,L Barros-Battesti and Knysak (1999) Peru Madre de Dios: Tambopata Agouti ? Need et al. (1991) ?: uncertain or unknown 1 Abbreviations of Brazilian states – AM: Amazonas; MG: Minas Gerais; PA: Pará; PR: Paraná; RJ: Rio de Janeiro; RO: Rondônia; RS: Rio Grande do Sul; SC: Santa Catarina; SP: São Paulo 2 F: female; N: nymph; L: larva

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