Systematics and biogeography of Anoura cultrata (Mammalia, Chiroptera, Phyllostomidae): a morphometric, niche modeling, and genetic perspective, with a taxonomic reappraisal of the genus.

The nectar-feeding bats of the genus Anoura are widely distributed in the Neotropics, but are most speciose in the Andes. Anoura cultrata is a rare mid-elevation bat occurring in South and Central America. It is thought to be one of the few bat species exemplifying a latitudinal cline in body size. We address three systematic and biogeographic questions: 1) is the geographic variation in A. cultrata continuous, as argued to justify its current monotypic status? 2) do ecogeographic barriers to dispersal affect such variation? and 3) how do the genetic divergence and biogeography of the species compare to those of other members of the genus? To answer these questions, we used morphometric analyses, ecological niche modeling, and DNA barcoding. We divided the samples of A. cultrata into six geographic groups, delimited by topographic depressions separating mountain systems. We did not find significant correlations between body size and the geographic coordinates within five groups. Therefore, we conclude that ecogeographic barriers to dispersal between the regions occupied by such groups influenced morphometric variation in A. cultrata, and that despite its general north to south reduction in body size, the species does not show continuous clinal variation. A recent phylogenetic study of the genus Anoura concluded that it contains seven valid species. Our DNA barcoding analysis and morphological examination indicated that at least 10 species should be recognized, including A. peruana distinct from A. geoffroyi, and A. aequatoris and A. luismanueli distinct from A. caudifer. Moreover, we show that Central and South American populations of A. cultrata differ from each other at least at the subspecific level, thus we respectively refer to them as A. cultrata cultrata and as A. c. brevirostrum. Similarly, we refer to Central American and Mexican populations of 'A. geoffroyi' as A. peruana lasiopyga, and to their South American counterparts as A. p. peruana. The range of the latter subspecies reaches northeastern Venezuela. The Andes from southern Colombia to northern Peru appear to be the ancestral range of the genus.

Genotyping-by-sequencing and ecological niche modeling illuminate phylogeography, admixture, and Pleistocene range dynamics in quaking aspen (Populus tremuloides).

Populus tremuloides is the widest-ranging tree species in North America and an ecologically important component of mesic forest ecosystems displaced by the Pleistocene glaciations. Using phylogeographic analyses of genome-wide SNPs (34,796 SNPs, 183 individuals) and ecological niche modeling, we inferred population structure, ploidy levels, admixture, and Pleistocene range dynamics of P. tremuloides, and tested several historical biogeographical hypotheses. We found three genetic lineages located mainly in coastal-Cascades (cluster 1), east-slope Cascades-Sierra Nevadas-Northern Rockies (cluster 2), and U.S. Rocky Mountains through southern Canadian (cluster 3) regions of the P. tremuloides range, with tree graph relationships of the form ((cluster 1, cluster 2), cluster 3). Populations consisted mainly of diploids (86%) but also small numbers of triploids (12%) and tetraploids (1%), and ploidy did not adversely affect our genetic inferences. The main vector of admixture was from cluster 3 into cluster 2, with the admixture zone trending northwest through the Rocky Mountains along a recognized phenotypic cline (Utah to Idaho). Clusters 1 and 2 provided strong support for the "stable-edge hypothesis" that unglaciated southwestern populations persisted in situ since the last glaciation. By contrast, despite a lack of clinal genetic variation, cluster 3 exhibited "trailing-edge" dynamics from niche suitability predictions signifying complete northward postglacial expansion. Results were also consistent with the "inland dispersal hypothesis" predicting postglacial assembly of Pacific Northwestern forest ecosystems, but rejected the hypothesis that Pacific-coastal populations were colonized during outburst flooding from glacial Lake Missoula. Overall, congruent patterns between our phylogeographic and ecological niche modeling results and fossil pollen data demonstrate complex mixtures of stable-edge, refugial locations, and postglacial expansion within P. tremuloides. These findings confirm and refine previous genetic studies, while strongly supporting a distinct Pacific-coastal genetic lineage of quaking aspen.

What is in a genus name? Conceptual and empirical issues preclude the proposed recognition of Callibella (Callitrichinae) as a genus.

In a recent article, Silva et al. (Zool Scr 47:133-143, 2018) proposed the relocation of the dwarf marmoset, Mico humilis, to the so far unrecognized genus Callibella. We contend that a taxonomic scheme that recognizes Callibella as if it were a valid genus is inadequately supported, and to some extent contradicted, by the ecological and morphological information provided by the authors. We discuss why the criterion of sympatry, invoked by Silva et al. to justify the recognition of Callibella at the genus level, is uninformative for taxonomic decisions above the species level. We also show that the morphological characteristics used by Silva et al. to separate Mico humilis from the other Mico are individually variable and present in every analyzed species of the genus. Moreover, we demonstrate that the Evolutionarily Distinct and Globally Endangered (EDGE) metric, employed by those authors to attempt to justify their taxonomic proposition, makes no sense in a taxonomic context. Conceptually, the use of autapomorphies and plesiomorphies to justify using Callibella goes against one of the main objectives of a meaningful classification, that is, to allow for all kinds of inferences based on previous observations (i.e., to be inductively projectible). Based on these arguments, we demonstrate that regarding Callibella as a subgenus of Mico is the most suitable way of making the Linnean taxonomy of marmosets congruent with the phylogenetic information available for the group.

Species delimitation based on diagnosis and monophyly, and its importance for advancing mammalian taxonomy.

A recently proposed taxonomic classification of extant ungulates sparked a series of publications that criticize the Phylogenetic Species Concept (PSC) claiming it to be a particularly poor species concept. These opinions reiteratively stated that (1) the two fundamental elements of the "PSC", i.e., monophyly and diagnosability, do not offer objective criteria as to where the line between species should be drawn; and (2) that extirpation of populations can lead to artificial diagnosability and spurious recognitions of species. This sudden eruption of criticism against the PSC is misleading. Problems attributed to the PSC are common to most approaches and concepts that modern systematists employ to establish species boundaries. The controversial taxonomic propositions that sparked criticism against the PSC are indeed highly problematic, not because of the species concept upon which they are based, but because no evidence (whatsoever) has become public to support a substantial portion of the proposed classification. We herein discuss these topics using examples from mammals. Numerous areas of biological research rest upon taxonomic accuracy (including conservation biology and biomedical research); hence, it is necessary to clarify what are (and what are not) the real sources of taxonomic inaccuracy.

Can Niche Modeling and Geometric Morphometrics Document Competitive Exclusion in a Pair of Subterranean Rodents (Genus Ctenomys) with Tiny Parapatric Distributions?

Species with similar ecological requirements coexisting in the same geographic region are prone to competitively exclude each other. Alternatively, they may coexist if character displacement acts to change the niche requirements of one or both species. We used two methodological approaches (ecological niche modeling [ENM] and geometric morphometrics) to test two hypotheses: given their behavioral, morphological, and ecological similarities, one species competitively excludes the other; and, character displacement enables their coexistence at two sites in which the species are known to occur in sympatry. The results from the ENM-based approach did not provide evidence for competitive exclusion; however, the morphometric analyses documented displacement in size of C. minutus. This result, suggests that C. minutus might exclude C. flamarioni from areas with softer soils and higher food availability. We stress the importance of using multiple methodological approaches when testing prediction of competitive exclusion. However, both methods had limited explanatory power given that the focal species possess truly peculiar distributions, being largely parapatric and restricted to narrow, small geographic areas with a strange distribution and there is a need to search for additional methods. We discuss the idiosyncrasy of the ENM-based approach when applied to organisms with subterranean habits.

A gene-tree test of the traditional taxonomy of American deer: the importance of voucher specimens, geographic data, and dense sampling.

The taxonomy of American deer has been established almost entirely on the basis of morphological data and without the use of explicit phylogenetic methods; hence, phylogenetic analyses including data for all of the currently recognized species, even if based on a single gene, might improve current understanding of their taxonomy. We tested the monophyly of the morphology-defined genera and species of New World deer (Odocoileini) with phylogenetic analyses of mitochondrial DNA sequences. This is the first such test conducted using extensive geographic and taxonomic sampling. Our results do not support the monophyly of Mazama, Odocoileus, Pudu, M. americana, M. nemorivaga, Od. hemionus, and Od. virginianus. Mazama contains species that belong to other genera. We found a novel sister-taxon relationship between "Mazama" pandora and a clade formed by Od. hemionus columbianus and Od. h. sitkensis, and transfer pandora to Odocoileus. The clade formed by Od. h. columbianus and Od. h. sitkensis may represent a valid species, whereas the remaining subspecies of Od. hemionus appear closer to Od. virginianus. Pudu (Pudu) puda was not found sister to Pudu (Pudella) mephistophiles. If confirmed, this result will prompt the recognition of the monotypic Pudella as a distinct genus. We provide evidence for the existence of an undescribed species now confused with Mazama americana, and identify other instances of cryptic, taxonomically unrecognized species-level diversity among populations here regarded as Mazama temama, "Mazama" nemorivaga, and Hippocamelus antisensis. Noteworthy records that substantially extend the known distributions of M. temama and "M." gouazoubira are provided, and we unveil a surprising ambiguity regarding the distribution of "M." nemorivaga, as it is described in the literature. The study of deer of the tribe Odocoileini has been hampered by the paucity of information regarding voucher specimens and the provenance of sequences deposited in GenBank. We pinpoint priorities for future systematic research on the tribe Odocoileini.

The mammalian faunas endemic to the Cerrado and the Caatinga.

We undertook a comprehensive, critical review of literature concerning the distribution, conservation status, and taxonomy of species of mammals endemic to the Cerrado and the Caatinga, the two largest biomes of the South American Dry-Diagonal. We present species accounts and lists of species, which we built with criteria that, in our opinion, yielded results with increased scientific rigor relative to previously published lists - e.g., excluding nominal taxa whose statuses as species have been claimed only on the basis of unpublished data, incomplete taxonomic work, or weak evidence. For various taxa, we provided arguments regarding species distributions, conservation and taxonomic statuses previously lacking in the literature. Two major findings are worth highlighting. First, we unveil the existence of a group of species endemic to both the Cerrado and the Caatinga (i.e., present in both biomes and absent in all other biomes). From the biogeographic point of view, this group, herein referred to as Caatinga-Cerrado endemics, deserves attention as a unit - just as in case of the Caatinga-only and the Cerrado-only endemics. We present preliminary hypotheses on the origin of these three endemic faunas (Cerrado-only, Caatinga-only, and Caatinga-Cerrado endemics). Secondly, we discovered that a substantial portion of the endemic mammalian faunas of the Caatinga and the Cerrado faces risks of extinction that are unrecognized in the highly influential Red List of Threatened Species published by the International Union for Conservation of Nature (IUCN). "Data deficient" is a category that misrepresents the real risks of extinction of these species considering that (a) some of these species are known only from a handful of specimens collected in a single or a few localities long ago; (b) the Cerrado and the Caatinga have been sufficiently sampled to guarantee collection of additional specimens of these species if they were abundant; (c) natural habitats of the Cerrado and the Caatinga have been substantially altered or lost in recent decades. Failures either in the design of the IUCN criteria or in their application to assign categories of extinction risks represent an additional important threat to these endemic faunas because their real risks of extinctions become hidden. It is imperative to correct this situation, particularly considering that these species are associated to habitats that are experiencing fast transformation into areas for agriculture, at an unbearable cost for biodiversity.

Photography-based taxonomy is inadequate, unnecessary, and potentially harmful for biological sciences.

The question whether taxonomic descriptions naming new animal species without type specimen(s) deposited in collections should be accepted for publication by scientific journals and allowed by the Code has already been discussed in Zootaxa (Dubois & Nemésio 2007; Donegan 2008, 2009; Nemésio 2009a-b; Dubois 2009; Gentile & Snell 2009; Minelli 2009; Cianferoni & Bartolozzi 2016; Amorim et al. 2016). This question was again raised in a letter supported by 35 signatories published in the journal Nature (Pape et al. 2016) on 15 September 2016. On 25 September 2016, the following rebuttal (strictly limited to 300 words as per the editorial rules of Nature) was submitted to Nature, which on 18 October 2016 refused to publish it. As we think this problem is a very important one for zoological taxonomy, this text is published here exactly as submitted to Nature, followed by the list of the 493 taxonomists and collection-based researchers who signed it in the short time span from 20 September to 6 October 2016.

Are we overestimating the niche? Removing marginal localities helps ecological niche models detect environmental barriers.

Correlative ecological niche models (ENMs) estimate species niches using occurrence records and environmental data. These tools are valuable to the field of biogeography, where they are commonly used to infer potential connectivity among populations. However, a recent study showed that when locally relevant environmental data are not available, records from patches of suitable habitat protruding into otherwise unsuitable regions (e.g., gallery forests within dry areas) can lead to overestimations of species niches and their potential distributions. Here, we test whether this issue obfuscates detection of an obvious environmental barrier existing in northern Venezuela - that of the hot and xeric lowlands separating the Península de Paraguaná from mainland South America. These conditions most likely promote isolation between mainland and peninsular populations of three rodent lineages occurring in mesic habitat in this region. For each lineage, we calibrated optimally parameterized ENMs using mainland records only, and leveraged existing habitat descriptions to assess whether those assigned low suitability values corresponded to instances where the species was collected within locally mesic conditions amidst otherwise hot dry areas. When this was the case, we built an additional model excluding these records. We projected both models onto the peninsula and assessed whether they differed in their ability to detect the environmental barrier. For the two lineages in which we detected such problematic records, only the models built excluding them detected the barrier, while providing additional insights regarding peninsular populations. Overall, the study reveals how a simple procedure like the one applied here can deal with records problematic for ENMs, leading to better predictions regarding the potential effects of the environment on lineage divergence.

In-solution hybridization for mammalian mitogenome enrichment: pros, cons and challenges associated with multiplexing degraded DNA.

Here, we present a set of RNA-based probes for whole mitochondrial genome in-solution enrichment, targeting a diversity of mammalian mitogenomes. This probes set was designed from seven mammalian orders and tested to determine the utility for enriching degraded DNA. We generated 63 mitogenomes representing five orders and 22 genera of mammals that yielded varying coverage ranging from 0 to >5400X. Based on a threshold of 70% mitogenome recovery and at least 10× average coverage, 32 individuals or 51% of samples were considered successful. The estimated sequence divergence of samples from the probe sequences used to construct the array ranged up to nearly 20%. Sample type was more predictive of mitogenome recovery than sample age. The proportion of reads from each individual in multiplexed enrichments was highly skewed, with each pool having one sample that yielded a majority of the reads. Recovery across each mitochondrial gene varied with most samples exhibiting regions with gaps or ambiguous sites. We estimated the ability of the probes to capture mitogenomes from a diversity of mammalian taxa not included here by performing a clustering analysis of published sequences for 100 taxa representing most mammalian orders. Our study demonstrates that a general array can be cost and time effective when there is a need to screen a modest number of individuals from a variety of taxa. We also address the practical concerns for using such a tool, with regard to pooling samples, generating high quality mitogenomes and detail a pipeline to remove chimeric molecules.

The Taxonomic Status of Mazama bricenii and the Significance of the Táchira Depression for Mammalian Endemism in the Cordillera de Mérida, Venezuela.

We studied the taxonomy and biogeography of Mazama bricenii, a brocket deer classified as Vulnerable by the IUCN, drawing on qualitative and quantitative morphology and sequences of the mitochondrial cytochrome-b gene. We used Ecological Niche Modeling (ENM) to evaluate the hypothesis that M. bricenii of the Venezuelan Cordillera de Mérida (CM) might have become isolated from populations of its putative sister species, Mazama rufina, in the Colombian Cordillera Oriental (CO). This hypothesis assumes that warm, dry climatic conditions in the Táchira Depression were unsuitable for the species. Our analyses did not reveal morphological differences between specimens geographically attributable to M. bricenii and M. rufina, and phylogenetic analyses of molecular data recovered M. bricenii nested within the diversity of M. rufina. These results indicate that M. bricenii should be regarded as a junior synonym of M. rufina. ENM analyses revealed the existence of suitable climatic conditions for M. rufina in the Táchira Depression during the last glacial maximum and even at present, suggesting that gene flow between populations in the CO and CM may have occurred until at least the beginning of the current interglacial period and may continue today. Because this pattern might characterize other mammals currently considered endemic to the CM, we examined which of these species match two criteria that we propose herein to estimate if they can be regarded as endemic to the CM with confidence: (1) that morphological or molecular evidence exists indicating that the putative endemic taxon is distinctive from congeneric populations in the CO; and (2) that the putative endemic taxon is restricted to either cloud forest or páramo, or both. Only Aepeomys reigi, Cryptotis meridensis, and Nasuella meridensis matched both criteria; hence, additional research is necessary to assess the true taxonomic status and distribution of the remaining species thought to be CM endemics.

No need to replace an "anomalous" primate (Primates) with an "anomalous" bear (Carnivora, Ursidae).

By means of mitochondrial 12S rRNA sequencing of putative "yeti", "bigfoot", and other "anomalous primate" hair samples, a recent study concluded that two samples, presented as from the Himalayas, do not belong to an "anomalous primate", but to an unknown, anomalous type of ursid. That is, that they match 12S rRNA sequences of a fossil Polar Bear (Ursusmaritimus), but neither of modern Polar Bears, nor of Brown Bears (Ursusarctos), the closest relative of Polar Bears, and one that occurs today in the Himalayas. We have undertaken direct comparison of sequences; replication of the original comparative study; inference of phylogenetic relationships of the two samples with respect to those from all extant species of Ursidae (except for the Giant Panda, Ailuropodamelanoleuca) and two extinct Pleistocene species; and application of a non-tree-based population aggregation approach for species diagnosis and identification. Our results demonstrate that the very short fragment of the 12S rRNA gene sequenced by Sykes et al. is not sufficiently informative to support the hypotheses provided by these authors with respect to the taxonomic identity of the individuals from which these sequences were obtained. We have concluded that there is no reason to believe that the two samples came from anything other than Brown Bears. These analyses afforded an opportunity to test the monophyly of morphologically defined species and to comment on both their phylogenetic relationships and future efforts necessary to advance our understanding of ursid systematics.