First record of Chiroderma improvisum Baker & Genoways, 1976 (Chiroptera: Phyllostomidae) from Saint Kitts, Lesser Antilles

: Chiroderma improvisum is a rare bat previously known only on the Caribbean Islands of Guadeloupe and Montserrat. We report the first recorded capture of C. improvisum on the island of Saint Kitts, 80 km northwest of Montserrat. Cytochrome b (cytB) gene analysis of the single captured specimen confirmed the identity of the bat as C. improvisum ; however, there is enough difference to indicate some population divergence, and possibly differentiation at the subspecific level among islands. We also report the first records of an ectoparasite, Periglischrus iheringi (Acarina: Spinturnicidae), from this bat.

One of the largest and rarest of bats in the Caribbean, Chiroderma improvisum Baker & Genoways, 1976, is known only from a handful of infrequent encounters on two islands located in the Lesser Antilles of the eastern Caribbean: Guadeloupe, where single bats were captured in 1974captured in , 2000captured in , and 2007captured in , and Montserrat, from which specimens were captured in 1978captured in , 1984captured in , and 2005 ( Figure 1) (Baker and Genoways 1976;Baker et al. 1978;Jones and Baker 1979;Larsen et al. 2007). Chiroderma improvisum was not reported from Saint Kitts in the catalog of bats from the island by Pedersen et al. (2005). Conservation and population status of this species are poorly understood and the bat is listed as vulnerable, due to restricted geographic range and habitat conversion by the increasing human population on these islands (Dávalos and Rodriguez 2008). Parasites and pathogens are unknown from this species. We report the capture of a single specimen of C. improvisum from the nation of Saint Kitts and Nevis, in the northern Lesser Antil-les. Saint Kitts Island consists of a composite volcanic island with some limestone uplift formations located 180 km northwest of Guadeloupe and 80 km northwest of Montserrat (Davis 1924).
The capture of this individual bat was incidental to a project conducted with Ross University School of Veterinary Medicine, which was examining bats on Saint Kitts for internal and external parasites. The project was approved by the Institutional Animal Care
The adult female ( Figure 2) of C. improvisum was netted near a house (17°19.222ʹ N, 062°43.615ʹ W using datum WGS84) on 28 November 2010. Our collection is the first with georeferenced data in the literature ( Figure 1). External measurements of forearm length 59 mm, tibia length 27 mm and mass of 4.7 g conform to what is known for the species (Table 1). It did not appear to be lactating or pregnant, as was typical of other species that we examined during late November. Pelage was dark grey/brown. There was some frosting throughout but more so ventrally. The dorsal pelage was purer brown and a thin white stripe was clearly evident. Chiroderma improvisum is the largest known Chiroderma with Chiroderma doriae being the only other similarsized species (Baker and Genoways 1976). Our specimen was within the morphometric range of C. improvisum from Guadeloupe and Montserrat (Baker and Genoways 1976;Jones and Baker 1979) and larger than the other Chiroderma species (Table 1). In addition, we used molecular tools to confirm the species identification.
Two parasitic mites in the family Spinturnicidae mites were collected, slide mounted, and identified using a stereoscopic microscope using keys by Furman (1966) and Morales-Malacara (2001). The specimens were deposited as vouchers GM/S<P>007 and GM/S<P>044 at Tomsk State University, Siberia, Russian Federation.
The cytochrome B gene has been sequenced for all named forms of Chiroderma (available on GenBank). Blood from the C. improvisum specimen was used for genetic analysis. The cytochrome B gene fragment was 98.6% (477/484 bp) identical to that from a C. and Use Committee of Ross University. There is little exposed fresh water on Saint Kitts, so we sought out streams, ponds, and swimming pools for mist netting sites. During November to December 2010, we captured and examined 92 bats (51 Artibeus jamaicensis Leach, 1921, 20 Ardops nichollsi (Thomas, 1891, 13 Molossus molossus (Pallas, 1766), 7 Brachyphylla cavernarum Gray, 1834, and 1 Chiroderma improvisum). Bats were identified to species, with sex, reproductive status, total body weight, and morphometric measurements recorded. Body measurements were made with calipers. We were not approved to kill C. improvisum as it is included on the IUCN Red List of Threatened Species, and the individual was photographed and released after blood and ectoparasite samples were taken.
Additionally, a thin blood film was prepared, stained with the CAMCO (Cambridge Diagnostic Products, Inc., Fort Lauderdale, FL) Stain Pak, and examined for the presence of blood parasites. Culture for bacteria in the genus Bartonella spp. was performed, using 50 µL of blood plated onto BHI/Sheep Blood Agar (Remel Products, Lenexa, KS), as previously described (Kosoy et al. 2010). DNA from blood was tested for Bartonella spp. using a sensitive real-time PCR assay (Reeves et al. 2007). Bartonella spp. are known to infect several species of tropical bats (Kosoy et al. 2010).
We obtained a 0.3 mL blood sample from the brachial vein of the C. improvisum. A 200-µL aliquot was used for DNA extraction, using a DNeasy kit (Qiagen, Valencia,   Figure  3); this analysis confirms that the specimen from Saint Kitts is C. improvisum, although it is slightly divergent from the previously accessioned sequence.
Male and female mites were collected from the C. improvisum from Saint Kitts. Both mites were identified as Periglischrus iheringi. These parasitic mites are found primarily on the wing membranes of Phyllostomidae (Furman 1966). No blood-borne parasites or Bartonella bacteria were detected in this bat, either by examination of a stained blood film, culture to isolate Bartonella spp., or by sensitive real-time PCR testing for Bartonella.
The C. improvisum was captured approximately 3 hours after sunset on 28 November 2010, in a 12 m × 2.6 m mist net set near an abandoned swimming pool that had collected 1.5 m of rainwater. Bats had to fly below ground level to get to the water. The water was clear but contained a small amount of debris and a large number of larval amphibians (tadpoles). Most of the trees on the island were not bearing fruit, but a large fruiting fig tree located 50 m from the pool was attracting large numbers of fruit bats. The net was intentionally placed between these the pool and tree.
External measurements, qualitative pelage characters, and mtDNA analysis confirm the identity of the Saint Kitts as Chiroderma improvisum. The external appearance and size of the bat is similar to C. improvisum found on Guadeloupe and Montserrat (Jones and Baker 1980), but a possible darker coloration and 1.4% variation in the genetic analysis could indicate separation from the previously identified populations. However, both our sample and the diversity of reference sequences in GenBank are quite limited. In our phylogenetic analysis, similar within-species divergence was noted between published sequences from specimens of Chiroderma villosum and Chiroderma doriae. Subspeciation of bats located on different islands of the Lesser Antilles has been reported for other endemic bat species, including A. nichollsi and B. cavernarum (Jones and Genoways 1973;Swanepoel and Genoways 1983). Additional DNA samples from C. improvisum from all three islands would be required to address this possibility; however, this bat is rarely captured, and all captures reported to date have included single specimens that were either released or deposited in museums.
The parasitic mite P. iheringi is a common ectoparasite of phyllostomids. It has not been reported on C. improvisum. Periglischrus iheringi is primarily found on wing membranes and probably feeds on both the host and other ectoparasites. It was reported on other species of Chiroderma by Smith (2012). Periglischrus iheringi was also collected from the other Phyllostomidae on Saint Kitts. It is not known to transmit pathogens to bats but has been associated with bacteria in the family Anaplasmataceae bacteria that might be an endosymbiont or possibly a pathogen (Reeves et al. 2006).  (Baker et al. 1994). Artibeus jamaicensis was included as an outgroup. Bootstrap replicates (n=1000) were performed, and bootstrap support is displayed as a precentage for each node. The scale bar indicates the expected number of nucleotide substitutions per site.