First records of ectoparasitic insects (Diptera: Hippoboscoidea) of bats in the department of Caldas, Colombia

In Colombia, the taxonomical understanding of ectoparasites (Diptera: Streblidae and Nycteribiidae) of bats is scarce, despite the high diversity of hosts. This fact reflects the lack of important information on the group and poses the need for studies that expand the knowledge on bat ectoparasites in different regions of the country. The aim of this study was to contribute pioneer knowledge on bat ectoparasitic insects in the department of Caldas, located in the Central Andes Mountain range of Colombia. We assessed 318 bats from four eco parks in the city of Manizales, and collected 234 ectoparasitic insects. We provide the first report of the family Streblidae for Caldas, with seven genera and eight species: Anastrebla caudiferae Wenzel, 1976, Anatrichobius scorzai Wenzel, 1966, Exastinion oculatum Wenzel, 1976, Megistopoda proxima (Séguy‚ 1926), Paratrichobius longicrus Ribeiro, 1907, Paraeuctenodes similis Wenzel, 1976, Trichobius longipes Rudow, 1871, and Trichobius tiptoni Wenzel, 1976. In addition, we report the species Basilia ferrisi Schuurmans Stekhoven, 1931 and Basilia sp. for the family Nycteribiidae. Key-Words. Insects; Ectoparasites; Bats; Caldas.

In Colombia, three orders, five families, 21 genera, and 85 bat ectoparasitic insect species are known to date (Bequaert, 1940;Guerrero, 1997;Marinkelle & Grose, 1981;Tarquino-Carbonell et al., 2015;Dick et al., 2016;Graciolli et al., 2016;Durán et al., 2017). The study by Bequaert (1940) was the first effort to study bat ectoparasitic insects from different localities of Colombia and Panamá. Later on, Marinkelle & Grose (1981) published an extensive list on Colombian bat ectoparasites of the orders Diptera, Hemiptera, and Siphonaptera. Recently, Tarquino-Carbonell et al. (2015) conducted a study on ectoparasites of chiropterofauna in the vereda Chorrillo, municipality of Ambalema, Tolima. Meanwhile, Durán et al. (2017) made the first report of Streblidae for the department of Sucre. Other contributions include the catalogues on Streblidae of the New World by Wenzel (1970) and Guerrero (1997), which include the species found in Colombia; in addition to the studies by Dick et al., 2016 andGraciolli et al., 2016 on Streblidae and Nycteribiidae species found in the country.
Despite the above, the information available for Colombia is scarce in comparison to other countries. Furthermore, there are no detailed studies on the taxonomy and/or composition of ectoparasitic species in several regions of the country. Considering the high diversity of bats in Colombia, this study aims to contribute to the understanding of ectoparasitic insects (Streblidae and Nycteribiidae) and their hosts, including the first reports for the department of Caldas, by studying bats and their ectoparasites in eco parks in the region.

Study area
Sampling was done in four eco parks of the municipality of Manizales, department of Caldas (Fig. 1). Ecoparque Alcázares-Arenillo: located in the southwestern region of the municipality of Manizales (05°04′N, 75°32′W), it has an extension of 70.55 Ha, altitude ranging between 1,730 m a.s.l. and 1,960 m a.s.l., and an average annual precipitation of 2,223 mm (Arango et al., 2007). This area is inhabited by species such as: Fraxinus chin-  (Arango et al., 2007). Jardín Botánico de la Universidad de Caldas: Located in southeast Manizales (05°04′N, 75°31′W), it has an extension of 7 Ha, at 2,150 m a.s.l., with an average precipitation over 1,800 mm (Corpocaldas, 2002 There are also 31.14 Ha in the process of secondary regeneration, following active intervention. This area was previously used for livestock farming, and was later destined for reforestation (Corpocaldas, 2002), with the inclusion of several species of Cecropia sp., Alnus acuminata, and Smallanthus sp.

Material collection and identification
Sampling was conducted from May to November 2014. Six 12 m × 2.5 m mist nets were utilized for bat capturing; the nets were relocated nightly to cover the greatest possible area. In total, 2,304 net hours were obtained from the four sampled zones. Morphometric data were collected from the bats according to Barquez (1987), along with additional data including sex, reproductive status, developmental stage, temperature, and weight. Bat identification was done according to the keys by Linares (1986), Timm et al., (1999); Muñoz (2001); Mantilla-Meluk & Baker (2006); Gardner (2007); Zurc & Velazco (2010); Solari et al. (2013). Several bats were taxidermized, including at least one individual per species (voucher), and eventually deposited in the Colección de Mamíferos del Museo de Historia Natural de la Universidad de Caldas (MHN-UC). Species collection was registered under permit number 215, emitted June 19 th , 2014 by the Corporación Autónoma Regional de Caldas (CORPOCALDAS).
Ectoparasites were collected from each live bat by screening the fur using a fine point tweezer and then ob-served under a stereoscopic magnifying glass (Graciolli & Carvalho, 2001). The specimens were deposited in Eppendorf tubes with 70% alcohol and later photographed in the laboratory using a Leica MC170 camera coupled to a Leica M205 stereomicroscope. The ectoparasites were identified using the taxonomic keys by Guimarães (1946), Guimarães & D'Andretta (1956), Wenzel et al. (1966), Wenzel (1970Wenzel ( , 1976, Guerrero (1995a, b), Autino et al. (1999), Graciolli & Carvalho (2001), and Dick & Miller (2010), among others, and the identifications were further confirmed by experts in the field.
Finally, the ectoparasites insects were deposited in the Colección Entomológica del Programa de Biología at the Universidad de Caldas (CEBUC) and the CMLA (Colección de Anexos de la Colección Mamíferos Lillo, de la Facultad de Ciencias Naturales e Instituto Miguel Lillo, Universidad Nacional de Tucumán, Argentina). Each Streblidae and Nycteribiidae specimen was catalogued by its corresponding number in the CMLA and deposited in the same collection.

Data analysis
Parasitism was assessed by prevalence values (P), which indicate the proportion or number of infested host bats in a population compared to the total number of bats examined. Furthermore, we calculated the intensity of infestation (I), also called parasitic load, which is defined as the number of parasites that live in or on a host (Wisnivesky, 2003). Finally, we estimated the specificity index (SE), which corresponds to the percentage of parasitism specificity of each ectoparasitic species in relation to each host bat species (Dick & Gettinger, 2005): Specificity index (SI) = ((Abundance of X ectoparasitic species in each host species/Total abundance of the ectoparasitic species) (100)).
Three hundred eighteen host bat specimens were captured ( The family Phyllostomidae was the most parasitized, specifically, by eight species of Streblidae; while the family Vespertilionidae was parasitized by three species, one belonging to Streblidae (A. scorzai) and two to Nycteribiidae (B. ferrisi and Basilia sp.). We found that 82%   of specimens were collected from Phyllostomidae, while the remaining 12% were extracted from Vespertilionidae.
The hosts with the highest number of ectoparasites were: C. brevicauda (n 79), A. lituratus (n 67), G. soricina (n 17), and D. bogotensis (n 12). We captured eight host species that were parasitized by more than one ectoparasitic species: C. brevicauda ( Table 1. Ectoparasitic insects species of Streblidae and Nycteribiidae (Diptera) and their host bats, with relative abundance (RA) and specificity index (SI). The specificity behavior to the host is also shown (HS): M: monoxene (found on only one host species); O: oligoxene (found on different bat hosts species, but of the same genus); Pl: pleioxene (found on different species and genera of host bats, but of the same family); Po: polyxene (found on different species and genera of host bats, and different families).   (Table 3). Bat parasitism prevalences and intensities in the four ecological parks show the highest prevalence for Los Alcázares eco park and highest intensity for El Recinto del Pensamiento (Table 4).

DISCUSSION
This study of ectoparasitic insects in bats of Caldas (Colombia) allows us to expand the understanding of the relations between ectoparasitic insects and their hosts. We found that the distribution and specificity of the ectoparasite is associated with the presence or absence of a given host species, and that the intensity and prevalence of insect parasitism in bats can vary according to the host species. For Colombia, there are reports of 19 genera and 73 species of Streblidae parasitizing Emballonuridae, Furipteridae, Molossidae, Mormoopidae, Natalidae, Noctilionidae, Phyllostomidae, and Vespertilionidae ; and two genera and eight species of Nycteribiidae for Vespertilionidae, Phyllostomidae, and Thyropteridae . The 10 ectoparasitic species found in this study comprise 14% of the total species reported for the country and they represent the first records of Streblidae and Nycteribiidae (Diptera) for the department of Caldas.
Exastinion oculatum has a more restricted distribution, which includes Colombia (Caldas in this study, Huila) and Panama (Wenzel, 1976;Guerrero, 1997;Dick et al., 2016). This species prefers hosts of the genus Anoura, specifically Anoura cultrata Handley, 1960 andA. geoffroyi (Guerrero, 1997;Dick et al., 2016). In this study, we report the species parasitizing C. brevicauda and G. soricina, potentially accidental hosts, a finding that broadens the range of known hosts for the species. It displays a pleioxene behavior in Caldas, since it expands hosts. In the rest of its distribution, however, it exhibits an oligoxene behavior.
Paratrichobius longicrus is a complex of species that parasitizes bats of the genera Artibeus and Platyrrhinus Saussure, 1860 (Wenzel et al., 1966). It is widely distributed in several countries, from Mexico in the north to Argentina in the south of the continent. In Colombia, it has been found in the departments of Antioquia, Meta, Tolima, Valle del Cauca, and Caldas in this study. The species has a wide range of hosts, mainly of the family Phyllostomidae, although Molossops planirostris Peters, 1865 and Molossops temminckii Burmeister, 1854 (Molossidae) have also been reported (Graciolli & Carvalho, 2001). Here, we report C. brevicauda, D. bogotensis, and Platyrrhinus helleri Peters, 1866 as hosts of P. longicrus. This species displays a polyxene behavior in its area of distribution, although, in Caldas, it is pleioxene.
Our findings showed that, in Caldas, each species behavior varies in terms of its hosts, which is likely due to a smaller sample area compared to the extensive distribution of each ectoparasite. As for the hosts, the family Phyllostomidae showed the highest species richness and abundance (11 spp.), encompassing 24% of the reported species for Caldas (48 spp.) (Castaño, 2012). This is followed by Vespertilionidae with eight species (61%) out of 13 species reported for Caldas and Molossidae with one species (20%) out of five reported for the department (Castaño, 2012). The forests in Caldas have un-dergone transformations from anthropic intervention and expansion of the agricultural frontier, among other causes, that affect the biodiversity of host bats and mammals in general (Castaño, 2012). This study on Streblidae and Nycteribiidae ectoparasites and their hosts in Caldas (Manizales) contributes a basis for future studies in other departments of Colombia that have not yet been sampled. Our study documents the enrichment of the diversity of host and ectoparasitic fauna in the country, in an ever-changing landscape.