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Traditionally, morphometric data have consisted of distances, angles, or ratios, and have been considered inappropriate for cladistic analyses. Recently, geometric morphometrics, based on homologous landmark point-coordinates, has provided a number of advantages over traditional morphometric data and methods, including the possibility that phylogenetically informative characters and character-states may be extracted and used in cladistic analyses. Using two data sets of 3-dimensional point coordinates collected from skulls of bats, we empirically evaluate this possibility. Partial warps were extracted from the point-coordinate matrix, and these were then re-coded by gap-coding, for use in the cladistic analyses. In the case of samples from Eidolon helvum populations (two mainland localities and four islands in the Gulf of Guinea), analyzing males and females separately, our analyses based on these data were unable to detect consistent phylogeographic patterns among the populations. In the case of samples from plecotine bat species, these analyses produced a consensus cladogram showing considerable concordance with an earlier cladistic analysis by us of this group. In both cases, our results reflect those of earlier studies (based on both morphologic and genetic data), suggesting that the data and analytic techniques described herein may have interesting utility in cladistic analyses.
The molecular identification of prey in faeces is an efficient non-invasive technique to study diet which requires both a satisfactory method of DNA extraction and the design of specific primers to selectively amplify prey's DNA. In this study we evaluated and compared the efficiency of two total DNA extraction methods and five primer pairs for the molecular identification of birds from scats, in particular from the giant noctule bat (Nyctalus lasiopterus). A modified DNA stool Mini Kit of Qiagen was tested against a modified silica method with a guanidinium thiocianate (GuSCN) applied after freezing and pulverizing the samples. We also checked two published vertebrate- and bird-generalist primer pairs and three bird-specific primer pairs designed by us (two pairs targeting the cytochrome b and one the cytochrome oxidase subunit I genes) that amplified shorter DNA fragments. The results show that pulverizing the scat remains before extraction was a very important step, presumably facilitating access to the well preserved DNA located inside the rachis of the feathers. The combination of our bird-specific designed primers showed a higher amplification rate than the generalist primers and allowed successful bird identification from the feathers excreted by the giant noctule bat in all the scat samples analyzed, independent of the preservation method used (dried and frozen). These methodological improvements will allow not only the study of the avian diet composition of the enigmatic giant noctule, but the extension of this methodology to other bird predators such as raptors.
In a survey of bats from La Rioja (Spain), several specimens of the mystacinus group were captured at different mountain localities. Genetic and morphologic analyses have revealed the presence of two lineages within this group in La Rioja. The lineages have been identified as corresponding to two different species: Myotis mystacinus sensu stricto and the recently described M. alcathoe. Both species were found using the same nocturnal refugia (caves) and the same forest habitats. This study extends the distribution of M. alcathoe west and southwards and adds a new mammal species to the Iberian fauna.
The Myotis nattereri species complex consists of an entangled group of Western Palaearctic bats characterized by fringing hairs along the rear edge of their uropatagium. Some members are relatively common while others are rare but all forms are morphologically very similar and their taxonomy is unresolved. Recent studies based on different molecular markers have shown that several major and unexpected lineages exist within this group of forest-dwelling bats. All the mitochondrial and nuclear markers tested to date have shown that these major lineages evolved as fully independent and coherent units and therefore each qualifies as distinct species. In the absence of proper morphological diagnosis, these lineages are informally referred to in the literature under different names. We explore here the external and craniodental variation of these lineages. Although all morphological measurements were overlapping between these lineages, we show that lineages can be completely discriminated in a multivariate morphometric space. Consistent with previous molecular reconstructions, these four major lineages represent two pairs of related species, each represented by a named species (Myotis nattereri s. str. and M. escalerai, respectively) and by unnamed forms (Myotis sp. A and Myotis sp. B, respectively). Herein we describe formally these two unnamed forms to clarify the taxonomy within this species complex. This new taxonomic view has important implication for the protection of these species, as three of the four taxa must now be considered as range-restricted species in need of conservation actions.
In July 2002, several bats of the genus Plecotus (Geoffroy, 1818) were captured at two localities of ‘Ordesa y Monte Perdido’ National Park (Central Pyrenees, Spain). They showed external characters that appeared intermediate between those of P. auritus and P. austriacus. Morphometric and genetic analysis have revealed that these long-eared bats should be ascribed to the recently recognized species P. macrobullaris. This study extends the distribution of this new species, whose western limit was considered, until now, to lie in the Alps, and adds a new mammal species to the Iberian fauna.
We investigate the contribution of the Iberian bat fauna to the cryptic diversity in Europe using mitochondrial (cytb and ND1) and nuclear (RAG2) DNA sequences. For each of the 28 bat species known for Iberia, samples covering a wide geographic range within Spain were compared to samples from the rest of Europe. In this general screening, almost 20% of the Iberian species showed important mitochondrial discontinuities (K2P distance values > 5%) either within the Iberian or between Iberian and other European samples. Within Eptesicus serotinus and Myotis nattereri, levels of genetic divergence between lineages exceeded 16%, indicating that these taxa represent a complex of several biological species. Other well-differentiated lineages (K2P distances between 5–10%) appeared within Hypsugo savii, Pipistrellus kuhlii and Plecotus auritus, suggesting the existence of further cryptic diversity. Most unsuspected lineages seem restricted to Iberia, although two have crossed the Pyrenees to reach, at least, Switzerland.
We use two mitochondrial DNA fragments with different substitution rates (cytochrome b gene and the control region) to address the following phylogeographic questions about western Palaearctic populations of the barbastelle bat (Barbastella barbastellus): 1) Do the Iberian populations of barbastelles show any genetic discontinuity associated with its present fragmented distribution?, 2) Is the Gibraltar Strait an effective barrier to gene flow for barbastelles? and 3) Is the subspecies from the Canary Islands genetically distinct from continental barbastelles? Our molecular survey shows that there is only a shallow genetic structure among populations of the Iberian Peninsula and Morocco, and probably, even across Europe until Thrace, although this last point needs to be confirmed. The Gibraltar Strait has not played any significant role as a biogeographic barrier to prevent the recent passage of European barbastelles to Morocco (or vice versa). Our phylogenetic reconstructions also confirm the taxonomic distinction of B. barbastellus guanchae as an endemic subspecies confined to the Canary Islands. The precise origin of this Canarian taxon is, nevertheless, still unclear as its mitochondrial lineage is distinct from any lineage found so far in Morocco and Iberia. This important genetic distinctness suggests either a relatively ancient colonization of the Canary Islands or that the source population of the founders have not yet been identified.
Due to a morphological uniformity typically shown by bent-winged bats, the taxonomic recognition of species and subspecies within the sole genus Miniopterus has been much questioned and revised. The situation and definition of the African species M. minor is particularly confused. This species is known from scattered and discontinuous records on both mainland coasts, Madagascar, São Tomé and Grand Comoro islands. The island forms have been included either within M. minor or considered as endemic species. To clarify their taxonomy, we compare mitochondrial DNA sequences of all the island forms with other related African Miniopterus. The genetic distances found in this study support a taxonomic recognition of the island forms at species level and the phylogenetic reconstructions based on these data suggest that the M. minor, as considered traditionally, is not a monophyletic group. The morphological similarities between the Miniopterus from São Tomé (West Africa) and Grand Comoro (East Africa) may reflect convergent evolution rather than a common ancestry.
We recorded the echolocation behavior of the molossid bat Mormopterus minutus, a species that uses a plastic call inventory. During its foraging activity, M. minutus searches for insects emitting rather long and narrow-band echolocation calls. Search call design however, can vary noticeably even in a continuous foraging pass. While echolocating in different flying conditions M. minutus uses several other call designs such as short CF, QCF, FM/QCF, FM and multi-harmonic FM, with or without harmonic overlap, and QCF/FM. Call plasticity characterizes most echolocation sequences, particularly in bats flying in open spaces. Call variation was also influenced by the presence of conspecifics. In those sequences containing echolocation calls from more than one bat, signals from different individuals were reliably identified. In contrast to other small molossids, the call designs in the echolocation inventory of M. minutus show a high level of plasticity. Our results suggest that M. minutus has combined the advantages of emitting several call designs, as shown by molossids, with the advantages of manipulating one signal design as shown by vespertilionids within the same sonar inventory.
The analysis of genetic diversity is routinely used to identify divergent intraspecific units and contribute to the knowledge base of biodiversity. In this study we used mitochondrial genetic diversity to propose three management units (MUs) for the Davy's nakedbacked bat (Pteronotus davyi), an insectivorous forest-dwelling species that is distributed in tropical and subtropical areas of America. We analyzed a 555 bp segment of the mitochondrial DNA (mtDNA) control region in 144 individuals from 18 localities spread across the species distribution range in Mexico. Our results demonstrated that the mitochondrial genetic diversity of P. davyi is distributed in three MUs, namely Gulf North, Pacific-Veracruz and Southeastern, with conservation priority, due to either the high mitochondrial genetic diversity or the high proportion of unique haplotypes, for the following populations: Playa de Oro, Arroyo del Bellaco and Catemaco in the Pacific-Veracruz region, and Agua Blanca, Sardina, Calakmul, Calcehtok and Kantemó from the Southeastern region. The Gulf North unit shows signs of the recent loss of genetic variability. These proposed conservation units could be considered a generalized model of conservation for other species of cave-dwelling bats that share the same habitats.
Due to its morphological conservatism, the American funnel-eared bats of the genus Natalus have had a complex taxonomic history. In Mexico only one species, N. stramineus, has traditionally been recognized. But recently a new endemic species, N. lanatus, was described in the region as a sympatric sibling species of N. stramineus. Natalus stramineus, with a geographic distribution extending from Mexico to Panama, was subsequently reassigned to N. mexicanus. In this study, we analyzed morphological and molecular characters to determine if two distinct species of funnel-eared bats (N. lanatus and N. mexicanus) occur in Mexico. We found that the proposed diagnostic morphological characters that separate the two taxa are not consistent and rarely differentiate between the two forms. Likewise, phylogenetic analyses do not support the separation of Natalus into two species in Mexico, but there is clearly geographic structure when all Mexican haplotypes are combined. The genetic distance between the haplotypes that could be ascribed to N. lanatus and N. mexicanus was typically within the range of conspecifics and not distinct species. To summarize, the results of both morphological and molecular analyses point to the presence of a single species of Natalus in Mexico.
One of the major challenges to understanding the evolution of Neotropical bats concerns our capacity to successfully scrutinize phylogenetic patterns associated with cases of cryptic species complexes. In this study Pteronotus parnellii is examined as a selected example of a known lineage of mormoopid bat that potentially contains several cryptic species. A samples of 452 individuals from 83 different localities, essentially covering its entire mainland distribution, was evaluated using two genetic markers: COI (mitochondrial) and DBY (nuclear) genes. The findings of this study strongly support the hypothesis of high genetic variability and identify at least six lineages within P. parnellii, some of which appear to be cryptic species.
Wagner's mustached bat (Pteronotus personatus) is an insectivorous bat distributed throughout America from Mexico to Brazil, which inhabits a range of habitats from rain forests to dry deciduous forests. There are two currently recognized species within the P. personatus complex, for which we examined 235 cytochrome oxidase I (COI) sequences and 138 cytochrome b (Cytb) sequences in order to explore its genetic variation in Mexico as well as in Central and South America. Our results reveal considerable differences in the genetic structure inside this species complex, indicating five genetic lineages: 1) Gulf of Mexico and the Mexican Pacific coastal plain to the Isthmus of Tehuantepec, 2) Southeastern Mexico, 3) Guatemala, 4) Guyana and Suriname-COI/Guyana and Venezuela-Cytb, and 5) Guyana, Suriname, French Guiana and Brazil. In addition, we used the isolation-with-migration coalescent method to estimate divergence times. The results indicate that vicariant events occurred roughly 1,624,000–2,450,000 years ago during the Early Pleistocene, wherein Central America was the center of two separate diversification processes, one toward Mexico and the other South America. The intraspecific lineages obtained for P. personatus demonstrate the need to reevaluate the species complex limits of this taxon.
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