Development of microsatellite markers in Myotis sodalis and cross-species amplification in M. gricescens, M. leibii, M. lucifugus, and M. septentrionalis

The Indiana bat (Myotis sodalis) is a highly endangered vespertilionid bat whose distribution is associated with limestone caves in the eastern United States. We present nine new polymorphic nuclear microsatellite markers for Myotis sodalis developed using an enriched library method. A total of 62 M. sodalis from two populations were used to estimate genetic diversity parameters. In M. sodalis, the number of alleles observed for each locus ranged from 17 to 48 alleles and expected heterozygosity ranged from 0.894 to 0.973. The 9 microsatellite markers were also tested on M. gricescens, M. leibii, M. lucifugus, and M. septentrionalis. These polymorphic microsatellite markers provide a valuable tool for investigating the population genetics of these species and will provide important genetic data useful for the conservation and recovery of the endangered Indiana bat.     

The status of the Japanese and East Asian bats of the genus Myotis (Vespertilionidae) based on mitochondrial sequences

The genus Myotis includes the largest number of species in the family Vespertilionidae (Chiroptera), and its members are distributed throughout most of the world. To re-evaluate the phylogenetic position of East Asian Myotis species with respect to Myotis species worldwide, we analyzed mitochondrial gene sequences of NADH dehydrogenase subunit 1 and cytochrome b from 24 East Asian individuals as well as 42 vespertilionid bats determined previously. The results suggest that: (1) some individuals having the same species name in Europe and Japan do not form a monophyletic clade, indicating that some bat species exhibit morphological convergence, (2) Japanese Myotis mystacinus forms a sister relationship with Myotis brandtii (Palaearctic), and both species are included in the American clade implying that an ancestor of these species originated in North America, and (3) the Black whiskered bat, Myotis pruinosus, is endemic to Japan and forms sister relationships with Myotis yanbarensis and Myotis montivagus collected from Okinawa (Japan) and Selangor (Malaysia), respectively, implying that M. pruinosus originated from the south. The systematics of Japanese and East Asian Myotis bats were revisited by considering their phylogenetic relationships. Our study provides the first extensive phylogenetic hypothesis of the genus Myotis that includes East Asian and Japanese species.     

Chromosome number reduction accompanied by extensive heterochromatin addition in the bat Glauconycteris beatrix (Mammalia; Chiroptera, Vespertilionidae)

The presumed ancestral karyotype of the bat family Vespertilionidae consists of 44 chromosomes with a fundamental number of autosomal arms (FNa) of 50. Previously, only two of the roughly 350 vespertilionid species have been reported with 2n lower than 26. In this paper we report the 2n = 22 karyotype of the African vespertilionid Glauconycteris beatrix which shows an X-autosome translocation and extended, paracentromeric, chromomycin-A3-positive heterochromatin.     

A family matter: conclusive resolution of the taxonomic position of the long-fingered bats, miniopterus

The long-fingered bats (Miniopterus sp.) are among the most widely distributed mammals in the world. However, despite recent focus on the systematics of these bats, their taxonomic position has not been resolved. Traditionally, they are considered to be sole members of Miniopterinae, 1 of 5 subfamilies within the largest family of bats, the Vespertilionidae. However, this classification has increasingly been called into question. Miniopterines differ extensively from other vespertilionids in numerous aspects of morphology, embryology, immunology, and, most recently, genetics. Recent molecular studies have proposed that the miniopterines are sufficiently distinct from vespertilionids that Miniopterinae should be elevated to full familial status. However, controversy remains regarding the relationship of the putative family, Miniopteridae to existing Vespertilionidae and to the closely related free-tailed bats, the Molossidae. We report here the first conclusive analysis of the taxonomic position of Miniopterus relative to all other bat families. We generated one of the largest chiropteran data sets to date, incorporating approximately 11 kb of sequence data from 16 nuclear genes, from representatives of all bat families and 2 Miniopterus species. Our data confirm the distinctiveness of Miniopterus, and we support previous recommendations to elevate these bats to full familial status. We estimate that they diverged from all other bat species approximately 49-38 MYA, which is comparable to most other bat families. Furthermore, we find very strong support from all phylogenetic methods for a sister group relationship between Miniopteridae and Vespertilionidae. The Molossidae diverged from these lineages approximately 54-43 MYA and form a sister group to the Miniopteridae-Vespertilionidae clade.     

广东丹霞山国家级自然保护区 蝙蝠物种多样性调查

于2017年至2018年对广东丹霞山国家级自然保护区的翼手目动物进行调查,主要采用日栖息地与夜栖息地、捕食区网捕等调查方法,共调查到翼手目5科13属23种,其中蝙蝠科(Vespertilionidae)9属15种,菊头蝠科(Rhinolophidae)1属4种,蹄蝠科(Hipposideridae)1属2种,假吸血蝠科(Megadermatidae)1属1种,犬吻蝠科(Molossidae)1属1种。从区系组成来看,以东洋界为主(19种),其次为广布种(3种),古北界仅1种。从栖息类型上看,分为洞栖型、树栖型及建筑物栖息型,以洞栖型为主(15种),建筑物栖息型其次(12种),树栖型最少(5种),但其中9个物种的栖息地类型同时包含了上述3种栖息类型中的2种。本研究在广东丹霞山发现中国蝙蝠新分布记录1种,为卡氏伏翼(Hypsugo cadornae);而中印鼠耳蝠(Myotis indochinensis)为中国分布的再次确认。     

Geographic variation in South American populations of Myotis nigricans () (Chiroptera, Vespertilionidae), with the description of two new species

The genus Myotis (Vespertilionidae, Myotinae) comprises a diverse group of small to large-sized vespertilionid bats that present a worldwide distribution. Twelve South American species are currently recognized. In this paper we evaluate the morphological and morphometric variation observed in South American populations of the most widespread species, Myotis nigricans. Against this background, two forms can be morphologically distinguished from M. nigricans and other known South American species. We describe these new species, documenting their diagnostic external and cranial characters by comparing them to other sympatric and cryptic species of South American Myotis. In addition, we provide an emended diagnosis of Myotis nigricans.     

Comparative chromosome painting of four Siberian Vespertilionidae species with Aselliscus stoliczkanus and human probes

Vespertilionidae is the largest chiropteran family that comprises species of different specialization and wide geographic distribution. Up to now, only a few vespertilionid species have been studied by molecular cytogenetic approaches. Here, we have investigated the karyotypic relationships of 4 Vespertilionidae species from Siberia by G-banding and comparative chromosome painting. Painting probes from Aselliscus stoliczkanus were used to establish interspecific homologous chromosomal segments in Myotis dasycneme (2n = 44), Murina hilgendorfi (2n = 44), Plecotus auritus (2n = 32), and Vespertilio murinus (2n = 38). Robertsonian translocations and a few inversions differentiated the karyotypes of the examined species. Painting of P. auritus karyotype with human probes revealed 3 previously undetected cryptic segments homologous to human chromosomes (Homo sapiens, HSA) 8, 15, and 19, respectively. As a consequence, the existence of 2 HSA 4 + 8 syntenies in the P. auritus karyotype has been proven. In addition, a pericentric inversion or centromere shift was revealed on the smallest metacentric P. auritus chromosome 16/17 using the HSA 16 probe explaining the different G-banding pattern in comparison to the homologous Myotis chromosome 16/17.     

Complete mitochondrial genome of the Hodgson's bat Myotis formosus (Mammalia, Chiroptera, Vespertilionidae)

The first complete mitochondrial genome (17,159 bp) of the Hodgson's bat Myotis formosus, which is an endangered species in South Korea, was sequenced and characterized. The genome included 13 protein-coding, 22 tRNA, and 2 rRNA genes, and 1 control region. It has high AT content and the same gene arrangement pattern as those of typical vertebrate mitochondrial genome. Within the control region, a 80 bp tandem repeat unit was iterated five times which was found in Domain I. It has been observed only in the vespertilionid bat group, and could contribute to identifying the species or genus, and also distinguishing it from other bat families.     

Characterization of dinucleotide microsatellite loci in big brown bats (Eptesicus fuscus), and their use in other North American vespertilionid bats

We describe the development of six microsatellite loci for big brown bats, Eptesicus fuscus. Microsatellite markers were isolated from a small insert genomic library, and tested on a population of 44 animals from the Pend d’Oreille Valley, in southeastern British Columbia, Canada. These six loci were highly variable, with 12–23 alleles per locus, and observed and expected heterozygosities of greater than 79.5%. The six primer sets, and three others that were not variable in E. fuscus, were tested on 11 other species in the families Vespertilionidae and Antrozoidae. All the tested loci amplified highly variable products in at least several other species.     

食鱼蝙蝠——大足鼠耳蝠初报

大足鼠耳蝠属蝙蝠科,为中国特有种类。食性分析和野外观察证实大足鼠耳蝠为食鱼蝙蝠。室内放养条件下发现大足鼠耳蝠一次进食能连续消耗5．0—10．0g麦穗鱼。本研究首次报道北京房山亦有大足鼠耳蝠分布,并介绍了其地理分布和保护现状。     

Multiplex panels of polymorphic microsatellite loci for two cryptic bat species of the genus Pipistrellus,developed by cross‐species amplification within the family Vespertilionidae

The paper presents multiplex panels of polymorphic microsatellites for two closely related cryptic species Pipistrellus pipistrellus and Pipistrellus pygmaeus. We tested the cross‐species amplification of 34 microsatellite loci, originally developed for five vespertilionid bat species. Ten and nine polymorphic loci in P. pipistrellus (mean number of alleles per locus = 10.5) and P. pygmaeus (8.1), respectively, in three multiplex polymerase chain reactions per species were amplified. All loci can be analysed in a single fragment analysis and can be used as markers to the study of evolution and the ecology of structured populations of socially living bats.     

West nile virus antibodies in bats from New Jersey and New York

Eighty-three serum samples were obtained from big brown (Eptesicus fuscus), little brown (Myotis lucifugus), and northern long-eared (Myotis septentriotalis) bats (Chiroptera: Vespertilionidae), from New Jersey and New York (USA) between July and October 2002. Samples were analyzed for neutralizing antibodies to West Nile virus (WNV) and St. Louis encephalitis (SLE) virus. One little brown bat and one northern long-eared bat tested positive for WNV neutralizing antibodies. No bats had antibodies to SLE virus. This was the first large-scale investigation of WNV infection in bats in New Jersey. Additional work is needed to determine the effects of WNV on bat populations.     

Is the Gibraltar Strait a barrier to gene flow for the bat Myotis myotis (Chiroptera: Vespertilionidae)?

Because of their role in limiting gene flow, geographical barriers like mountains or seas often coincide with intraspecific genetic discontinuities. Although the Strait of Gibraltar represents such a potential barrier for both plants and animals, few studies have been conducted on its impact on gene flow. Here we test this effect on a bat species (Myotis myotis) which is apparently distributed on both sides of the strait. Six colonies of 20 Myotis myotis each were sampled in southern Spain and northern Morocco along a linear transect of 1350 km. Results based on six nuclear microsatellite loci reveal no significant population structure within regions, but a complete isolation between bats sampled on each side of the strait. Variability at 600 bp of a mitochondrial gene (cytochrome b) confirms the existence of two genetically distinct and perfectly segregating clades, which diverged several million years ago. Despite the narrowness of the Gibraltar Strait (14 km), these molecular data suggest that neither males, nor females from either region have ever reproduced on the opposite side of the strait. Comparisons of molecular divergence with bats from a closely related species (M. blythii) suggest that the North African clade is possibly a distinct taxon warranting full species rank. We provisionally refer to it as Myotis cf punicus Felten 1977, but a definitive systematic understanding of the whole Mouse-eared bat species complex awaits further genetic sampling, especially in the Eastern Mediterranean areas.     

Cryptic diversity in European bats

Different species of bat can be morphologically very similar. In order to estimate the amount of cryptic diversity among European bats we screened the intra- and interspecific genetic variation in 26 European vespertilionid bat species. We sequenced the DNA of subunit 1 of the mitochondrial protein NADH dehydrogenase (ND1) from several individuals of a species, which were sampled in a variety of geographical regions. A phylogeny based on the mitochondrial (mt) DNA data is in good agreement with the current classification in the family. Highly divergent mitochondrial lineages were found in two taxa, which differed in at least 11% of their ND1 sequence. The two mtDNA lineages in Plecotus austriacus correlated with the two subspecies Plecotus austriacus austriacus and Plecotus austriacus kolombatovici. The two mtDNA lineages in Myotis mystacinus were partitioned among two morphotypes. The evidence for two new bat species within Europe is discussed. Convergent adaptive evolution might have contributed to the morphological similarity among distantly related species if they occupy similar ecological niches. Closely related species may differ in their ecology but not necessarily in their morphology. On the other hand, two morphologically clearly different species (Eptesicus serotinus and Eptesicus nilssonii) were found to be genetically very similar. Neither morphological nor mitochondrial DNA sequence analysis alone can be guaranteed to identify species.     

A faunal survey of streblid flies (Diptera: Streblidae) associated with bats in Paraguay

An extensive survey of the ectoparasites infesting bats in Paraguay provides information regarding the taxonomy and host distribution of streblid bat flies at a geographic interface between subtropical and temperate habitats. Five families of bats representing 45 species, including Molossidae (5 genera and 15 species), Natalidae (1 genus and 1 species), Phyllostomidae (11 genera and 15 species), Noctilionidae (1 genus and 2 species), and Vespertilionidae (4 genera and 12 species) were collected from 24 localities across Paraguay and sampled for ectoparasites. In total, 2,467 bat flies were collected, representing 11 genera and 31 nominal species of Streblidae, of which 6 genera and 24 species are new records for Paraguay. No streblids were collected from vespertilionid bats; 23 species infested phyllostomids, 6 species noctilionids, 1 species a natalid, and 1 species molossids. Streblid bat flies were highly specific to certain host groups and individual host species, and their geographic distributions closely followed those of their host bats. Of 31 streblid species surveyed, 27 were monoxenous (i.e., associated with a single host species), and 4 were stenoxenous (i.e., associated with a group of phylogenetically related hosts). The number of streblid species is greatly reduced in the Chaco region west of the Paraguay River, largely because of the lack of phyllostomid host bats.     

A new species of Myotis Kaup, 1829 (Chiroptera, Vespertilionidae) from Ecuador

A new species of bat in the genus Myotis (Vespertilionidae, Myotinae) is described from the Chocó ecoregion on the western slope of the Andes in northwestern Ecuador. The genus Myotis comprises a diverse group of small to large-sized vespertilionid bats distributed worldwide. Twelve South American species are recognized currently, 6 of which occur in Ecuador. Morphological relationships among the new species and the other 12 South American species of Myotis were examined using 15 cranial and 5 external characters. Both qualitative and quantitative analyses found the new species to be distinct. As an aid to future identifications, we provide a key to the Ecuadorian species of Myotis.     

Comparative Analysis of the Amino Acid Spectrum of Blood Plasma in Chiroptera (<Emphasis Type="Italic">Vespertilio murinus</Emphasis> L., 1758 and <Emphasis Type="Italic">Myotis dasycneme</Emphasis> B., 1825) in the Fauna of the Ural Mountains

The article presents the results of a new comparative analysis of free amino acids in the blood plasma of representatives of insectivorous Chiroptera (Mammalia: Vespertilionidae) in the fauna of the Ural Mountains: the pond bat (Myotis dasycneme Boie, 1825) and the parti-colored bat (Vespertilio murinus Linnaeus, 1758). This is the first study to show the species variability of free amino acids in resident and migratory species of bats from different ecosystems of the Ural Region.     

Pectoralis muscle morphology in the little brown bat,Myotis lucifugus: A non-convergence with birds

Recent studies of muscle architecture demonstrate that many mammalian muscles are composed of short, interdigitating fibers. In addition, the avian pectoralis, a muscle capable of producing high frequency oscillations has been shown to possess a serially arranged pattern of muscle endplate in all sizes of birds studied. The pectoralis muscle of the little brown bat, Myotis lucifugus (Chiroptera: Vespertilionidae), is composed of fairly uniform fibers that span the length of the muscle and is characterized by a zone of motor endplates within the middle third of the muscle. The homogeneous fiber architecture of the bat pectoralis muscle is in contrast to the serial arrangement of endplates (and presumably muscle muscle fibers) in the avian pectoralis in species equivalent in size to Myotis. The short fiber organization and motor endplate pattern observed in most birds is thus not a requisite design for flying vertebrates. © 1994 Wiley-Liss, Inc.     

Microsatellite markers for the endemic New Zealand long-tailed bat (Chalinolobus tuberculatus)

The New Zealand long-tailed bat (Chalinolobus tuberculatus) is an endangered species which is now absent from many habitats where they were previously observed. However, population genetics studies have been hampered by the lack of reliable genetic markers. We have developed 13 microsatellite markers, and at least eight of these are suitable for population studies. Nine of these markers also cross-react with the closely related Australian species C. gouldii. These markers may prove of general utility in the genus Chalinolobus.     

Phylogeography of the greater horseshoe bat, Rhinolophus ferrumequinum: contrasting results from mitochondrial and microsatellite data

Phylogeographical studies are typically based on haplotype data, occasionally on nuclear markers such as microsatellites, but rarely combine both. This is unfortunate because the use of markers with contrasting modes of inheritance and rates of evolution might provide a more accurate and comprehensive understanding of a species' history. Here we present a detailed study of the phylogeography of the greater horseshoe bat, Rhinolophus ferrumequinum , using 1098 bp of the mitochondrial ND2 gene from 45 localities from across its Palaearctic range to infer population history. In addition, we re-analysed a large microsatellite data set available for this species and compared the results of both markers to infer population relationships and the historical processes influencing them. We show that mtDNA, the most popular marker in phylogeography studies, yielded a misleading result, and would have led us to conclude erroneously that a single expansion had taken place in Europe. Only by combining the mitochondrial and microsatellite data sets are we able to reconstruct the species' history and show two colonization events in Europe, one before the Last Glacial Maximum (LGM) and one after it. Combining markers also revealed the importance of Asia Minor as an ancient refugium for this species and a source population for the expansion of the greater horseshoe bat into Europe before the LGM.