South American Bats Research Articles

Bat blunders

Bat blunders

Whole-genome analysis reveals contrasting relationships among nuclear and mitochondrial genomes between three sympatric bat species.

Understanding mechanisms involved in speciation can be challenging, especially when hybridization or introgression blurs species boundaries. In bats resolving relationships of some closely related groups has proven difficult due subtle interspecific variation both in morphometrics and molecular datasets. The endemic South American Histiotus bats, currently considered a sub-genus of Eptesicus, harbor unresolved phylogenetic relationships and of those is a trio consisting of two closely related species: Eptesicus (Histiotus) macrotus and E. (H.) montanus, and their relationship with a third, E. (H.) magellanicus. The three sympatric species bear marked resemblance to each other, but can be differentiated morphologically. Furthermore, previous studies have been unable to differentiate the species from each other at a molecular level. In order to disentangle the phylogenetic relationships of these species, we examined the differentiation patterns and evolutionary history of the three Eptesicus (H.) species from Chile at the whole genome level. The nuclear DNA statistics between the species suggest strong gene flow and recent hybridization between E. (H.) montanus and E. (H.) macrotus, whereas E. (H.) magellanicus shows a higher degree of isolation. In contrast, mitochondrial DNA shows a closer relationship between E. (H.) magellanicus and E. (H.) montanus. Opposing patterns in mtDNA and nuclear markers are often due to differences in dispersal, and here it could be both as result of isolation in refugia during the last glacial maximum and female philopatry and male-biased dispersal. In conclusion, this study shows the importance of both the nuclear and mitochondrial DNA in resolving phylogenetic relationships and species histories.

Novel Borrelia Genotypes from Brazil Indicate a New Group of Borrelia spp. Associated with South American Bats.

The bacterial genus Borrelia comprises vector-borne spirochetes that have been classified into three major groups: the relapsing fever group (RFG), the Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner sensu lato group (Bbsl), and the reptile-monotreme group (RMG). All three groups have been associated mainly with ticks and wild animals, especially rodents, birds, and reptiles. Here, we searched for Borrelia infection among 99 vampire bats [Desmodus rotundus (É. Geoffroy)] (Chiroptera: Phyllostomidae) from the Brazilian semiarid region. Through molecular investigation of bat internal organs, haplotypes of a potentially novel Borrelia organism were detected in 5% (5/99) of the bats. Borrelia DNA was detected in the liver, blood, spleen, kidney and brain, suggesting a systemic infection. Phylogenetic analyses inferred from partial sequences of the borrelial rrs and flaB genes indicated that the vampire bat-associated Borrelia sp. of this study form a monophyletic group with a newly reported Borrelia associated with a Colombia bat, distinct from the three main currently recognized groups of Borrelia spp., Bbsl, RFG, and RMG. These novel bat-associated Borrelia spp. from South America might have arisen through an independent event along the borrelial evolutionary history, since previous molecular reports of Borrelia organisms in bats or bat-associated ticks from Africa, Europe, and North America were all classified in the RFG.

Molecular Evidence of Orthomyxovirus Presence in Colombian Neotropical Bats.

The Orthomyxoviridae family includes the genera Influenzavirus, Isavirus, Quaranjavirus, and Thogotovirus. In turn, Influenzavirus can be classified into four types: α, β, γ, and δ (Formerly A, B, C, and D), from which Alphainfluenzavirus (AIV) has the broadest host range, including birds, mammals, reptiles, and amphibians. Additionally, AIV has shown global epidemiological relevance owing to its pandemic potential. The epidemiological relevance of Chiropteran due to its multiple functional characteristics makes them ideal reservoirs for many viral agents. Recently, new influenza-like subtypes have been reported in Neotropical bats, but little is known about the relevance of bats as natural reservoirs of influenza viruses. Therefore, the current study aimed to determine the presence of AIV and new influenza-like subtypes in South American bats. For a better understanding of the drivers and interactions between AIV and bats, we used molecular assays with different gene targets (i.e., M, NP, and PB1) to identify AIV in New World bats. A housekeeping gene (CytB) PCR was used to check for nucleic acid preservation and to demonstrate the bat-origin of the samples. A total of 87 free-living bats belonging to 25 different species of the families Phyllostomidae and Vespertilionidae were collected in Casanare, Colombia. As a result, this study found seven AIV-positive bat species, three of them reported for the first time as AIV prone hosts. Neither of the AIV-like analyzed samples were positive for H17N10/H18/N11 subtypes. Although additional information is needed, the presence of a completely new or divergent AIV subtype in neotropical bats cannot be discarded. Collectively, the results presented here expand the epidemiological knowledge and distribution of AIV in neotropical free-ranging bats and emphasize the need to continue studying these viruses to establish the role they could play as a threat to animal and public health.

A new species of Long-eared Brown Bat of the genus <i>Histiotus</i> (Chiroptera) and the revalidation of <i>Histiotus colombiae</i>

The South American bats of the genus Histiotus comprise between four and eight species, but their taxonomy has been controversial and the limits between species and their distribution are not well understood. In Colombia, Histiotus humboldti and H. montanus colombiae have been recorded, but undescribed species has been suggested. We evaluated the species richness and distribution of Colombian Histiotus using morphological, molecular, and acoustic traits. Our results evidence three species in Colombia, the two previously recorded taxa and a new species from the Cordillera Central of Colombia and northern Ecuador that we describe here. We also revalidated H. colombiae as a full species. H. humboldti is widely distributed in the Colombian and Ecuadorean Andes and can be sympatric with the other two species. H. colombiae is restricted to the Colombian Cordillera Oriental. Finally, we highlight the potential hidden diversity within Histiotus in the Peruvian and Bolivian Andes, the need to resolve the evolutionary relationships of the genus, and its implications to the understanding of the processes that have structured the Andean mammal fauna.

Isolation and establishment of skin-derived and mesenchymal cells from south American bat Artibeus planirostris (Chiroptera – Phyllostomidae)

Animal models represent a crucial tool for biological research, so the establishment of new cultures is fundamental for the discovery of new therapies and the understanding of mechanisms of cell development in the most diverse animals. Here, we report the successful establishment of two new primary cell cultures derived from a South American bat (Artibeus planirostris). The establishment of a new bat culture can help in the investigation of new zoonoses since bats have been proposed as carriers of these diseases. We evaluated the chromosomal stability of cells from different passages. Primary cultures were collected from ear tissues and bone marrow of A. planirostris. Cultures were expanded, and osteogenic and adipogenic inductions were conducted for 21 days. For osteogenic differentiation, the medium was supplemented with 0.1 μM dexamethasone, 3 mM β-glycerophosphate, and 10 μM L-ascorbic acid 2-phosphate. For adipogenic differentiation, the medium was supplemented with 5 μM rosiglitazone, 0.4 μM insulin, 0.1 mM indomethacin, and 0.1 μM dexamethasone. After the induction period, the cells were stained with Alizarin Red to assess osteogenic differentiation and Oil Red O to assess adipogenic differentiation. We observed the appearance of lipid droplets in adipocytes and the extracellular deposition of calcium matrix by osteocytes, indicating that bone marrow-derived cells and skin-derived cells of A. planirostris could successfully differentiate into these lineages. Also, the number of chromosomes remained stable for both primary cultures during passages 2, 4, 6, and 8.

A Preliminary Study of the Virome of the South American Free-Tailed Bats (Tadarida brasiliensis) and Identification of Two Novel Mammalian Viruses.

Bats provide important ecosystem services as pollinators, seed dispersers, and/or insect controllers, but they have also been found harboring different viruses with zoonotic potential. Virome studies in bats distributed in Asia, Africa, Europe, and North America have increased dramatically over the past decade, whereas information on viruses infecting South American species is scarce. We explored the virome of Tadarida brasiliensis, an insectivorous New World bat species inhabiting a maternity colony in Rosario (Argentina), by a metagenomic approach. The analysis of five pooled oral/anal swab samples indicated the presence of 43 different taxonomic viral families infecting a wide range of hosts. By conventional nucleic acid detection techniques and/or bioinformatics approaches, the genomes of two novel viruses were completely covered clustering into the Papillomaviridae (Tadarida brasiliensis papillomavirus type 1, TbraPV1) and Genomoviridae (Tadarida brasiliensis gemykibivirus 1, TbGkyV1) families. TbraPV1 is the first papillomavirus type identified in this host and the prototype of a novel genus. TbGkyV1 is the first genomovirus reported in New World bats and constitutes a new species within the genus Gemykibivirus. Our findings extend the knowledge about oral/anal viromes of a South American bat species and contribute to understand the evolution and genetic diversity of the novel characterized viruses.

Two New Species and New Records of Mites of the Genus Parichoronyssus (Acari: Macronyssidae) From South American Bats (Chiroptera), With a Key to the Known Species of the Genus.

Derived from exhaustive search of mites of the genus Parichoronyssus associated with South American Bats, we found two new species associated with Phyllostomid, Emballonurid, and Noctiniolid bats: Parichoronyssus alexanderfaini n. sp. associated with Rhinophylla pumilio Peters; Parichoronyssus gettingeri n. sp. associated with Rhynchonycteris naso Wied-Newied, and Noctilio leporinus Linneo. Herein we give the description of those new species, and additionally is included several new records for Parichoronyssus from the region, as well as providing a key to the 11 species of Parichoronyssus.

Unexpected Functional Divergence of Bat Influenza Virus NS1 Proteins.

ABSTRACTRecently, two influenza A virus (FLUAV) genomes were identified in Central and South American bats. These sequences exhibit notable divergence from classical FLUAV counterparts, and functionally, bat FLUAV glycoproteins lack canonical receptor binding and destroying activity. Nevertheless, other features that distinguish these viruses from classical FLUAVs have yet to be explored. Here, we studied the viral nonstructural protein NS1, a virulence factor that modulates host signaling to promote efficient propagation. Like all FLUAV NS1 proteins, bat FLUAV NS1s bind double-stranded RNA and act as interferon antagonists. Unexpectedly, we found that bat FLUAV NS1s are unique in being unable to bind host p85β, a regulatory subunit of the cellular metabolism-regulating enzyme, phosphoinositide 3-kinase (PI3K). Furthermore, neither bat FLUAV NS1 alone nor infection with a chimeric bat FLUAV efficiently activates Akt, a PI3K effector. Structure-guided mutagenesis revealed that the bat FLUAV NS1-p85β interaction can be reengineered (in a strain-specific manner) by changing two to four NS1 residues (96L, 99M, 100I, and 145T), thereby creating a hydrophobic patch. Notably, ameliorated p85β-binding is insufficient for bat FLUAV NS1 to activate PI3K, and a chimeric bat FLUAV expressing NS1 with engineered hydrophobic patch mutations exhibits cell-type-dependent, but species-independent, propagation phenotypes. We hypothesize that bat FLUAV hijacking of PI3K in the natural bat host has been selected against, perhaps because genes in this metabolic pathway were differentially shaped by evolution to suit the unique energy use strategies of this flying mammal. These data expand our understanding of the enigmatic functional divergence between bat FLUAVs and classical mammalian and avian FLUAVs.IMPORTANCE The potential for novel influenza A viruses to establish infections in humans from animals is a source of continuous concern due to possible severe outbreaks or pandemics. The recent discovery of influenza A-like viruses in bats has raised questions over whether these entities could be a threat to humans. Understanding unique properties of the newly described bat influenza A-like viruses, such as their mechanisms to infect cells or how they manipulate host functions, is critical to assess their likelihood of causing disease. Here, we characterized the bat influenza A-like virus NS1 protein, a key virulence factor, and found unexpected functional divergence of this protein from counterparts in other influenza A viruses. Our study dissects the molecular changes required by bat influenza A-like virus NS1 to adopt classical influenza A virus properties and suggests consequences of bat influenza A-like virus infection, potential future evolutionary trajectories, and intriguing virus-host biology in bat species.

Discovery of African bat polyomaviruses and infrequent recombination in the large T antigen in the Polyomaviridae.

Bat species represent natural reservoirs for a number of high-consequence human pathogens. The present study investigated the diversity of polyomaviruses (PyVs) in Zambian insectivorous and fruit bat species. We describe the complete genomes from four newly proposed African bat PyV species employing the recently recommended criteria provided by the Polyomaviridae Study Group of the International Committee on Taxonomy of Viruses. A comprehensive phylogenetic and recombination analysis was performed to determine genetic relationships and the distribution of recombination events in PyV from mammalian and avian species. The novel species of PyV from Zambian bats segregated with members of the genera Alphapolyomavirus and Betapolyomavirus, forming monophyletic clades with bat and non-human primate PyVs. Miniopterus schreibersii polyomavirus 1 and 2 segregated in a clade with South American bat PyV species, Old World monkey and chimpanzee PyVs and Human polyomavirus 13 (New Jersey PyV). Interestingly, the newly described Egyptian fruit bat PyV, tentatively named Rousettus aegyptiacus polyomavirus 1, had the highest nucleotide sequence identity to species of PyV from Indonesian fruit bats, and Rhinolophus hildebrandtii polyomavirus 1 was most closely related to New World monkey PyVs. The distribution of recombination events in PyV genomes was non-random: recombination boundaries existed in the intergene region between VP1 and LTAg and also at the 3' end of VP2/3 in the structural genes, whereas infrequent recombination was present within the LTAg gene. These findings indicate that recombination within the LTAg gene has been negatively selected against during polyomaviral evolution and support the recent proposal for taxonomic classification based on LTAg to define novel PyV species.

New Rickettsia species in soft ticks Ornithodoros hasei collected from bats in French Guiana

In French Guiana, located on the northeastern coast of South America, bats of different species are very numerous. The infection of bats and their ticks with zoonotic bacteria, especially Rickettsia species, is so far unknown. In order to improve knowledge of these zoonotic pathogens in this French overseas department, the presence and diversity of tick-borne bacteria was investigated with molecular tools in bat ticks.In the beginning of 2013, 32 bats were caught in Saint-Jean-du-Maroni, an area close to the coast of French Guiana, and the ticks of these animals were collected. A total of 354 larvae of Argasidae soft ticks (Ornithodoros hasei) from 12 bats (Noctilio albiventris) were collected and 107 of them were analysed. DNA was extracted from the samples and quantitative real-time PCR was carried out to detect Rickettsia spp., Bartonella spp., Borrelia spp. and Coxiella burnetii. All tested samples were negative for Bartonella spp., Borrelia spp. and Coxiella burnetii. Rickettsia DNA was detected in 31 (28.9%) ticks. An almost entire (1118 base pairs long) sequence of the gltA gene was obtained after the amplification of some positive samples on conventional PCR and sequencing. A Bayesian tree was constructed using concatenated rrs, gltA, ompA, ompB, and gene D sequences. The study of characteristic sequences shows that this Rickettsia species is very close (98.3–99.8%) genetically to R. peacockii. Nevertheless, the comparative analysis of sequences obtained from gltA, ompA, ompB, rrs and gene D fragments demonstrated that this Rickettsia is different from the other members of the spotted fever group. The sequences of this new species were deposited in GenBank as Candidatus Rickettsia wissemanii. This is the first report showing the presence of nucleic acid of Rickettsia in Ornithodoros hasei ticks from South American bats.

Zoogeography of South American Forest-Dwelling Bats: Disjunct Distributions or Sampling Deficiencies?

Many forest-dwelling bats are purported to be widespread in South America, although records are scant from the vast diagonal belt of dry ecosystems that straddles the continent, implying possible sampling deficiencies. Here, we investigate this possibility in the case of four species of bat (Centronycteris maximiliani, Lampronycteris brachyotis, Peropteryx kappleri and Trinycteris nicefori), evaluating whether their disjunct present-day distributions reflect their true zoogeographic characteristics or the subsampling of intermediate zones. We use environmental niche modelling (ENM) in an ensemble approach, combining four different modeling techniques, and using niche descriptors based on climatic and remote sensing data, to estimate the potential distribution of the four species. The models indicate that all four species have disjunct distributions in the Amazon and Atlantic forest biomes. The one possible exception is P. kappleri, which the models indicated might potentially occur in humid forest enclaves in western Brazil and eastern Bolivia. The present-day distribution of the species may date back to the Plio-Pleistocene, when the forested biomes of South America were more extensive and connected. Further studies of different chiropteran lineages may provide additional insights into the historic processes of faunal interchange between the Amazon and Atlantic forest biomes.

Bat fauna of Mato Grosso do Sul, southwestern Brazil

Bats have been increasingly studied in the last 15 years in Mato Grosso do Sul, and several records were not yet considered in reviews of South American bat distributions. Here, we present the bat species and their distributions in Mato Grosso do Sul based mainly on data compilation from literature, but also on complementary information from zoological collections, and our and colleagues’ unpublished records. We found 74 species of bats within 42 genera and seven families already reported in Mato Grosso do Sul. Bat species in this state represent 44% of the Brazilian’s bat species (≅ 169) and 7% of the world’s bat richness (≅ 1120). Phyllostomidae (42) and Molossidae (17) were the richest families. Four species formerly cited for Mato Grosso do Sul are not supported by our compilation, and other 15 species recorded in the vicinity are listed as potential occurrences in this state. We additionally found controversial traits for specimens of Platyrrhinus helleri, and report Eumops dabbenei for the first time in Brazil. Most species were recorded in the regions of Cerrado (60) or Pantanal (57) in Mato Grosso do Sul, but only 16 in the Atlantic Forest. Records of Phyllostomidae species were mostly found in Cerrado and those of Molossidae, in Pantanal. Records in Mato Grosso do Sul determine edges of distribution for at least 22 species of South American bats. The overall known chiropteran fauna of Mato Grosso do Sul is highly diverse and new findings are expected through additional surveys.

Geographic and Potential Distribution of a Poorly Known South American Bat,Histiotus macrotus(Chiroptera: Vespertilionidae)

The vespertilionid bat Histiotus macrotus occurs in western Argentina, central regions of Chile and south of Argentina and Chile, and it may be also present in Bolivia and southern Peru. In this work, we analyzed the geographic and potential distribution of a poorly known species of South American bat. As a tool, environmental niche modeling has been used to study the distributional patterns of species and more recently, taxonomic boundaries of cryptic species. We used MaxEnt v 3.3.e, Worldclim database and a vegetation map, covering the entire area of species' occurrence. We registered 64 localities from Argentina (43), Chile (10), Peru (8) and Bolivia (5). We divided recording localities in different datasets according to several taxonomic schemes, and analyzed potential distribution models separately (i.e., all known records; Argentina-Chile; Peru) in five different models. Models including all known localities showed a disjoint distribution, with two basic core areas of high predictive values, one in NW Argentina and another in southern Chile and SW Argentina separated by the South American Arid Diagonal. A third area appeared in Atacama and Sechura deserts in the models that included Peruvian and Bolivian localities. Model including only Peruvian localities showed the opposite pattern, with high predictive values only in arid environments from southern Peru. We interpreted that localities correctly assigned to H. macrotus belong to a taxonomic complex distributed in two contrasting areas, each one inhabited by a different taxon: 1) Bolivia and NW Argentina and 2) S and central Chile and SW Argentina. Given the systematic uncertainty of Histiotus, these two forms might be sister species or may not share an immediate common ancestor within the genus. Further, we consider that the specimens from localities referred to H. macrotus from southern Peru should be revised. These alternatives await a comprehensive molecular phylogenetic analysis of Histiotus.

Checklist of the helminth parasites of South American bats.

Although the Chiroptera represents a significant proportion (c.20%) of the mammalian fauna and South America has the highest diversity of bat species, only about a third of the known species in this region have had helminth parasites reported from them. This work represents the first comprehensive checklist of the helminth parasites (nematodes, acanthocephalans, trematodes and cestodes) of South American bats. The data were extracted from more than 120 references and are presented as a key to each group of parasites down to the generic level, with an indication of how the bats become infected, accompanied by a list of the species recorded for each genus. This is followed, in tabular form, by parasite-host and host-parasite checklists. The parasite-host list also includes their geographical distribution in South America (at the country level) and site data, plus the references in which the parasite records occur. The host-parasite list is arranged according to the classification of the hosts. In all, c.370 host-parasite associations are recorded, involving 114 nominal species of helminths from 92 named chiropteran taxa.

No virological evidence for an influenza A - like virus in European bats.

New members of the influenza A virus genus have been detected recently in bats from South America. By molecular investigations, using a generic real-time RT-PCR (RT-qPCR) that detects all previously known influenza A virus subtypes (H1-H16) and a newly developed RT-qPCR specific for the South American bat influenza-like virus of subtype H17, a total of 1571 samples obtained from 1369 individual bats of 26 species from Central Europe were examined. No evidence for the occurrence of such influenza viruses was found. Further attempts towards a more comprehensive evaluation of the role of bats in the ecology and epidemiology of influenza viruses should be based on more intense monitoring efforts. However, given the protected status of bats, not only in Europe, such activities need to be embedded into existing pathogen-monitoring programs.

Novel polyomaviruses in South American bats and their relationship to other members of the family Polyomaviridae

Bats are the natural reservoir of a variety of viruses, including a polyomavirus (PyV) from a North American brown bat. We investigated 163 spleen samples from 22 bat species from French Guiana for the presence of PyVs. In total, we detected 25 PyV-positive animals belonging to nine different bat species. Phylogenetic analysis was performed on the genomes of eight representative PyVs, and showed that the bat PyVs form three distinct lineages within the genus Orthopolyomavirus and are genetically different from the previously described North American bat virus. Interestingly, two lineages cluster with PyVs found in chimpanzees, orangutans and gorillas. In addition, one group of bat PyVs is genetically related to the human Merkel cell polyomavirus.

Morphometric and morphological variation in Myotis simus Thomas (Chiroptera, Vespertilionidae), with an appraisal of the identity of Myotis guaycuru Proença based on the analysis of the type material

Twelve species are recognized in the South American bat genus Myotis Kaup (Vespertilionidae, Myotinae), with several nominal forms currently regarded as synonyms, among them Myotis guaycuru Proença, 1943. Its holotype, so far the only specimen assigned to the species, has not been examined in recent taxonomic reviews. To address the taxonomic status of M. guaycuru, we located and redescribed its holotype and compared it to representatives and/or descriptions of all South American species in the genus. Qualitative traits, namely the plagiopatagium attached at ankles, the short and wooly fur and the lingually displaced P3, unambiguously assign the holotype of M. guaycuru to Myotis simus Thomas (1901). The analysis of cranial variation and pelage color across a wide geographical range of M. simus reveals morphometric and morphological discontinuity between Bolivian and Amazonian/Peruvian samples, the latter including topotypes of M. simus. The holotype of M. guaycuru was found to be morphometrically and morphologically more similar to these Amazonian samples than to the geographically nearer Bolivian sample, preventing the use of this nominal form to refer to the Bolivian population if its distinction suggested by morphometric analyses is confirmed by the analyses of other character systems.

Partial molecular characterization of alphaherpesviruses isolated from tropical bats

Herpesviruses have previously been isolated from African and South-American bats. Recently, herpesviruses detected from European insectivorous bats (family Vespertilionidae) were classified molecularly as betaherpesviruses and gammaherpesviruses. In the current study, we performed PCR analyses targeting the UL30 catalytic subunit region of the DNA polymerase gene of the African and South American herpesviruses and new Malagasy and Cambodian herpesviruses isolated from bats, especially frugivorous bats from the families Pteropodidae and Phyllostomidae. The sequences obtained from the amplified products indicated that these isolates belonged to the genus Simplexvirus of the subfamily Alphaherpesvirinae. These results extend the taxonomic range of bat herpesviruses with the description of four members in the subfamily Alphaherpesvirinae. Furthermore, these data confirm and extend the geographical distribution of herpesvirus in bats to three more continents (Africa, South America and Asia) and indicate the presence of these viruses in frugivorous bats of the families Pteropodidae and Phyllostomidae.

Detection and Phylogenetic Analysis of Group 1 Coronaviruses in South American Bats

Bat coronaviruses (Bt-CoVs) are thought to be the precursors of severe acute respiratory syndrome coronavirus. We detected Bt-CoVs in 2 bat species from Trinidad. Phylogenetic analysis of the RNA-dependent RNA polymerase gene and helicase confirmed them as group 1 coronaviruses.