The Role of Bats and Relationships As Reservoirs of Zoonotic Viruses and the Origin of New Coronaviruses
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Four Species in One: Multigene Analyses Reveal Phylogenetic
Published by Associazione Teriologica Italiana Volume 29 (1): 111–121, 2018 Hystrix, the Italian Journal of Mammalogy Available online at: http://www.italian-journal-of-mammalogy.it doi:10.4404/hystrix–00017-2017 Research Article Four species in one: multigene analyses reveal phylogenetic patterns within Hardwicke’s woolly bat, Kerivoula hardwickii-complex (Chiroptera, Vespertilionidae) in Asia Vuong Tan Tu1,2,3,4,∗, Alexandre Hassanin1,2,∗, Neil M. Furey5, Nguyen Truong Son3,4, Gábor Csorba6 1Institut de Systématique, Evolution, Biodiversité (ISYEB), UMR 7205 MNHN CNRS UPMC, Muséum national d’Histoire naturelle, Case postale N°51–55, rue Buffon, 75005 Paris, France 2Service de Systématique Moléculaire, UMS 2700, Muséum national d’Histoire naturelle, Case postale N°26–43, rue Cuvier, 75005 Paris, France 3Institute of Ecology and Biological Resources, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet road, Cau Giay district, Hanoi, Vietnam 4Graduate University of Science and Technology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet road, Cau Giay district, Hanoi, Vietnam 5Fauna & Flora International, Cambodia Programme, 19 Street 360, Boeng Keng Kang 1, Chamkarmorn, Phnom Penh, Cambodia 6Department of Zoology, Hungarian Natural History Museum, Baross u. 13., H-1088, Budapest, Hungary Keywords: Abstract Kerivoulinae Asia We undertook a comparative phylogeographic study using molecular, morphological and morpho- phylogeography metric approaches to address systematic issues in bats of the Kerivoula hardwickii complex in Asia. taxonomy Our phylogenetic reconstructions using DNA sequences of two mitochondrial and seven nuclear cryptic species genes reveal a distinct clade containing four small-sized species (K. hardwickii sensu stricto, K. depressa, K. furva and Kerivoula sp. -
Bat Coronavirus in the Western Indian Ocean
bioRxiv preprint doi: https://doi.org/10.1101/742866; this version posted September 4, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Title: Bat coronavirus phylogeography in the western Indian Ocean 2 Running title: Bat coronavirus in the western Indian Ocean 3 Authors: Léa Joffrin*, Steven M. Goodman, David A. Wilkinson, Beza Ramasindrazana, Er- 4 wan Lagadec, Yann Gomard, Gildas Le Minter, Andréa Dos Santos, M. Corrie Schoeman, Ra- 5 jendraprasad Sookhareea, Pablo Tortosa, Simon Julienne, Eduardo S. Gudo, Patrick Mavingui 6 and Camille Lebarbenchon 7 8 Author affiliations : 9 Université de La Réunion, UMR Processus Infectieux en Milieu Insulaire Tropical (PIMIT) 10 INSERM 1187, CNRS 9192, IRD 249, Sainte-Clotilde, La Réunion, France (L. Joffrin, B. Ra- 11 masindrazana, E. Lagadec, Y. Gomard, G. Le Minter, D.A. Wilkinson, P. Tortosa, P. Mavingui, 12 C. Lebarbenchon) 13 Association Vahatra, Antananarivo, Madagascar (S.M. Goodman, B. Ramasindrazana) 14 Field Museum of Natural History, Chicago, USA (S.M. Goodman) 15 Veterinary Faculty, Eduardo Mondlane University, Maputo, Mozambique (A. Dos Santos) 16 School of Life Sciences, University of Kwa-Zulu Natal, Kwa-Zulu Natal, South Africa (M.C. 17 Schoeman) 18 National Parks and Conservation Service, Réduit, Mauritius (R. Sookhareea) 19 Seychelles Ministry of Health, Victoria, Mahe, Seychelles (S. Julienne) 20 Instituto Nacional de Saúde, Maputo, Mozambique (E.S. Gudo). 21 22 *Corresponding author: [email protected] 23 UMR PIMIT, 2 rue Maxime Rivière, 97490 Sainte-Clotilde, Reunion Island, France. 24 Tel: +262 262 93 88 00 1 bioRxiv preprint doi: https://doi.org/10.1101/742866; this version posted September 4, 2019. -
Mammalian Skull Heterochrony Reveals Modular Evolution and a Link Between Cranial Development and Brain Size
ARTICLE Received 31 Oct 2013 | Accepted 11 Mar 2014 | Published 4 Apr 2014 DOI: 10.1038/ncomms4625 OPEN Mammalian skull heterochrony reveals modular evolution and a link between cranial development and brain size Daisuke Koyabu1,2, Ingmar Werneburg1, Naoki Morimoto3, Christoph P.E. Zollikofer3, Analia M. Forasiepi1,4, Hideki Endo2, Junpei Kimura5, Satoshi D. Ohdachi6, Nguyen Truong Son7 & Marcelo R. Sa´nchez-Villagra1 The multiple skeletal components of the skull originate asynchronously and their develop- mental schedule varies across amniotes. Here we present the embryonic ossification sequence of 134 species, covering all major groups of mammals and their close relatives. This comprehensive data set allows reconstruction of the heterochronic and modular evolution of the skull and the condition of the last common ancestor of mammals. We show that the mode of ossification (dermal or endochondral) unites bones into integrated evolutionary modules of heterochronic changes and imposes evolutionary constraints on cranial heterochrony. How- ever, some skull-roof bones, such as the supraoccipital, exhibit evolutionary degrees of freedom in these constraints. Ossification timing of the neurocranium was considerably accelerated during the origin of mammals. Furthermore, association between developmental timing of the supraoccipital and brain size was identified among amniotes. We argue that cranial heterochrony in mammals has occurred in concert with encephalization but within a conserved modular organization. 1 Palaeontological Institute and Museum, University of Zu¨rich, Karl Schmid-Strasse 4, Zu¨rich 8006, Switzerland. 2 The University Museum, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan. 3 Anthropological Institute and Museum, University of Zu¨rich, Winterthurerstrasse 190, Zu¨rich 8057, Switzerland. -
Desmodus Rotundus) Blood Feeding
toxins Article Vampire Venom: Vasodilatory Mechanisms of Vampire Bat (Desmodus rotundus) Blood Feeding Rahini Kakumanu 1, Wayne C. Hodgson 1, Ravina Ravi 1, Alejandro Alagon 2, Richard J. Harris 3 , Andreas Brust 4, Paul F. Alewood 4, Barbara K. Kemp-Harper 1,† and Bryan G. Fry 3,*,† 1 Department of Pharmacology, Biomedicine Discovery Institute, Faculty of Medicine, Nursing & Health Sciences, Monash University, Clayton, Victoria 3800, Australia; [email protected] (R.K.); [email protected] (W.C.H.); [email protected] (R.R.); [email protected] (B.K.K.-H.) 2 Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Cuernavaca, Morelos 62210, Mexico; [email protected] 3 Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4067, Australia; [email protected] 4 Institute for Molecular Biosciences, University of Queensland, St Lucia, QLD 4072, Australia; [email protected] (A.B.); [email protected] (P.F.A.) * Correspondence: [email protected] † Joint senior authors. Received: 20 November 2018; Accepted: 2 January 2019; Published: 8 January 2019 Abstract: Animals that specialise in blood feeding have particular challenges in obtaining their meal, whereby they impair blood hemostasis by promoting anticoagulation and vasodilation in order to facilitate feeding. These convergent selection pressures have been studied in a number of lineages, ranging from fleas to leeches. However, the vampire bat (Desmondus rotundus) is unstudied in regards to potential vasodilatory mechanisms of their feeding secretions (which are a type of venom). This is despite the intense investigations of their anticoagulant properties which have demonstrated that D. -
Exposure of Humans Or Animals to Sars-Cov-2 from Wild, Livestock, Companion and Aquatic Animals Qualitative Exposure Assessment
ISSN 0254-6019 Exposure of humans or animals to SARS-CoV-2 from wild, livestock, companion and aquatic animals Qualitative exposure assessment FAO ANIMAL PRODUCTION AND HEALTH / PAPER 181 FAO ANIMAL PRODUCTION AND HEALTH / PAPER 181 Exposure of humans or animals to SARS-CoV-2 from wild, livestock, companion and aquatic animals Qualitative exposure assessment Authors Ihab El Masry, Sophie von Dobschuetz, Ludovic Plee, Fairouz Larfaoui, Zhen Yang, Junxia Song, Wantanee Kalpravidh, Keith Sumption Food and Agriculture Organization for the United Nations (FAO), Rome, Italy Dirk Pfeiffer City University of Hong Kong, Hong Kong SAR, China Sharon Calvin Canadian Food Inspection Agency (CFIA), Science Branch, Animal Health Risk Assessment Unit, Ottawa, Canada Helen Roberts Department for Environment, Food and Rural Affairs (Defra), Equines, Pets and New and Emerging Diseases, Exotic Disease Control Team, London, United Kingdom of Great Britain and Northern Ireland Alessio Lorusso Istituto Zooprofilattico dell’Abruzzo e Molise, Teramo, Italy Casey Barton-Behravesh Centers for Disease Control and Prevention (CDC), One Health Office, National Center for Emerging and Zoonotic Infectious Diseases, Atlanta, United States of America Zengren Zheng China Animal Health and Epidemiology Centre (CAHEC), China Animal Health Risk Analysis Commission, Qingdao City, China Food and Agriculture Organization of the United Nations Rome, 2020 Required citation: El Masry, I., von Dobschuetz, S., Plee, L., Larfaoui, F., Yang, Z., Song, J., Pfeiffer, D., Calvin, S., Roberts, H., Lorusso, A., Barton-Behravesh, C., Zheng, Z., Kalpravidh, W. & Sumption, K. 2020. Exposure of humans or animals to SARS-CoV-2 from wild, livestock, companion and aquatic animals: Qualitative exposure assessment. FAO animal production and health, Paper 181. -
On the Coronaviruses and Their Associations with the Aquatic Environment and Wastewater
water Review On the Coronaviruses and Their Associations with the Aquatic Environment and Wastewater Adrian Wartecki 1 and Piotr Rzymski 2,* 1 Faculty of Medicine, Poznan University of Medical Sciences, 60-812 Pozna´n,Poland; [email protected] 2 Department of Environmental Medicine, Poznan University of Medical Sciences, 60-806 Pozna´n,Poland * Correspondence: [email protected] Received: 24 April 2020; Accepted: 2 June 2020; Published: 4 June 2020 Abstract: The outbreak of Coronavirus Disease 2019 (COVID-19), a severe respiratory disease caused by betacoronavirus SARS-CoV-2, in 2019 that further developed into a pandemic has received an unprecedented response from the scientific community and sparked a general research interest into the biology and ecology of Coronaviridae, a family of positive-sense single-stranded RNA viruses. Aquatic environments, lakes, rivers and ponds, are important habitats for bats and birds, which are hosts for various coronavirus species and strains and which shed viral particles in their feces. It is therefore of high interest to fully explore the role that aquatic environments may play in coronavirus spread, including cross-species transmissions. Besides the respiratory tract, coronaviruses pathogenic to humans can also infect the digestive system and be subsequently defecated. Considering this, it is pivotal to understand whether wastewater can play a role in their dissemination, particularly in areas with poor sanitation. This review provides an overview of the taxonomy, molecular biology, natural reservoirs and pathogenicity of coronaviruses; outlines their potential to survive in aquatic environments and wastewater; and demonstrates their association with aquatic biota, mainly waterfowl. It also calls for further, interdisciplinary research in the field of aquatic virology to explore the potential hotspots of coronaviruses in the aquatic environment and the routes through which they may enter it. -
Inferring the Ecological Niche of Bat Viruses Closely Related to SARS-Cov-2 Using Phylogeographic Analyses of Rhinolophus Specie
www.nature.com/scientificreports OPEN Inferring the ecological niche of bat viruses closely related to SARS‑CoV‑2 using phylogeographic analyses of Rhinolophus species Alexandre Hassanin1,4*, Vuong Tan Tu2,4, Manon Curaudeau1 & Gabor Csorba3 The Severe Acute Respiratory Syndrome coronavirus 2 (SARS‑CoV‑2) is the causal agent of the coronavirus disease 2019 (COVID‑19) pandemic. To date, viruses closely related to SARS‑CoV‑2 have been reported in four bat species: Rhinolophus acuminatus, Rhinolophus afnis, Rhinolophus malayanus, and Rhinolophus shameli. Here, we analysed 343 sequences of the mitochondrial cytochrome c oxidase subunit 1 gene (CO1) from georeferenced bats of the four Rhinolophus species identifed as reservoirs of viruses closely related to SARS‑CoV‑2. Haplotype networks were constructed in order to investigate patterns of genetic diversity among bat populations of Southeast Asia and China. No strong geographic structure was found for the four Rhinolophus species, suggesting high dispersal capacity. The ecological niche of bat viruses closely related to SARS‑CoV‑2 was predicted using the four localities in which bat viruses were recently discovered and the localities where bats showed the same CO1 haplotypes than virus‑positive bats. The ecological niche of bat viruses related to SARS‑CoV was deduced from the localities where bat viruses were previously detected. The results show that the ecological niche of bat viruses related to SARS‑CoV2 includes several regions of mainland Southeast Asia whereas the ecological niche of bat viruses related to SARS‑CoV is mainly restricted to China. In agreement with these results, human populations in Laos, Vietnam, Cambodia, and Thailand appear to be much less afected by the COVID‑19 pandemic than other countries of Southeast Asia. -
Mammalian Skull Heterochrony Reveals Modular Evolution and a Link Between Cranial Development and Brain Size
Title Mammalian skull heterochrony reveals modular evolution and a link between cranial development and brain size Koyabu, Daisuke; Werneburg, Ingmar; Morimoto, Naoki; Zollikofer, Christoph P. E.; Forasiepi, Analia M.; Endo, Author(s) Hideki; Kimura, Junpei; Ohdachi, Satoshi D.; Truong Son, Nguyen; Sánchez-Villagra, Marcelo R. Nature Communications, 5(3625) Citation https://doi.org/10.1038/ncomms4625 Issue Date 2014-04-04 Doc URL http://hdl.handle.net/2115/62700 Rights(URL) https://creativecommons.org/licenses/by-nc-sa/3.0/ Type article File Information ncomms4625.pdf Instructions for use Hokkaido University Collection of Scholarly and Academic Papers : HUSCAP ARTICLE Received 31 Oct 2013 | Accepted 11 Mar 2014 | Published 4 Apr 2014 DOI: 10.1038/ncomms4625 OPEN Mammalian skull heterochrony reveals modular evolution and a link between cranial development and brain size Daisuke Koyabu1,2, Ingmar Werneburg1, Naoki Morimoto3, Christoph P.E. Zollikofer3, Analia M. Forasiepi1,4, Hideki Endo2, Junpei Kimura5, Satoshi D. Ohdachi6, Nguyen Truong Son7 & Marcelo R. Sa´nchez-Villagra1 The multiple skeletal components of the skull originate asynchronously and their develop- mental schedule varies across amniotes. Here we present the embryonic ossification sequence of 134 species, covering all major groups of mammals and their close relatives. This comprehensive data set allows reconstruction of the heterochronic and modular evolution of the skull and the condition of the last common ancestor of mammals. We show that the mode of ossification (dermal or endochondral) unites bones into integrated evolutionary modules of heterochronic changes and imposes evolutionary constraints on cranial heterochrony. How- ever, some skull-roof bones, such as the supraoccipital, exhibit evolutionary degrees of freedom in these constraints. -
Ultraconserved Elements Are Novel Phylogenomic Markers That Resolve Placental Mammal Phylogeny When Combined with Species Tree Analysis
Downloaded from genome.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press Ultraconserved elements are novel phylogenomic markers that resolve placental mammal phylogeny when combined with species tree analysis John E. McCormack,1,8 Brant C. Faircloth,2 Nicholas G. Crawford,3 Patricia Adair Gowaty,4,5 Robb T. Brumfield1,6 & Travis C. Glenn7 1 Museum of Natural Science, Louisiana State University, Baton Rouge, LA 70803; 2 Department of Ecology and Evolutionary Biology, University of California, Los Angeles, CA 90095; 3 Department of Biology, Boston University, Boston, MA 02215; 4 Smithsonian Tropical Research Institute, MRC 0580-11 Unit 9100, Box 0948, DPO, AA 34002-9998, USA; 5 Institute of the Environment, University of California, Los Angeles, CA 90095; 6 Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803; 7 Department of Environmental Health Science, University of Georgia, Athens, GA 30602 Running Title: Ultraconserved elements fuel species-tree phylogenomics Keywords: phylogenomics, coalescence 8 Corresponding author: Moore Laboratory of Zoology, Occidental College, 1600 Campus Rd., Los Angeles, CA 90041; E-mail: [email protected]; Tel: 734-358-6886 Page 1 Downloaded from genome.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press ABSTRACT Phylogenomics offers the potential to fully resolve the Tree of Life, but increasing genomic coverage also reveals conflicting evolutionary histories among genes, demanding new analytical strategies for elucidating a single history of life. Here, we outline a phylogenomic approach using a novel class of phylogenetic markers derived from ultraconserved elements and flanking DNA. Using species-tree analysis that accounts for discord among hundreds of independent loci, we show that this class of marker is useful for recovering deep-level phylogeny in placental mammals. -
Index of Handbook of the Mammals of the World. Vol. 9. Bats
Index of Handbook of the Mammals of the World. Vol. 9. Bats A agnella, Kerivoula 901 Anchieta’s Bat 814 aquilus, Glischropus 763 Aba Leaf-nosed Bat 247 aladdin, Pipistrellus pipistrellus 771 Anchieta’s Broad-faced Fruit Bat 94 aquilus, Platyrrhinus 567 Aba Roundleaf Bat 247 alascensis, Myotis lucifugus 927 Anchieta’s Pipistrelle 814 Arabian Barbastelle 861 abae, Hipposideros 247 alaschanicus, Hypsugo 810 anchietae, Plerotes 94 Arabian Horseshoe Bat 296 abae, Rhinolophus fumigatus 290 Alashanian Pipistrelle 810 ancricola, Myotis 957 Arabian Mouse-tailed Bat 164, 170, 176 abbotti, Myotis hasseltii 970 alba, Ectophylla 466, 480, 569 Andaman Horseshoe Bat 314 Arabian Pipistrelle 810 abditum, Megaderma spasma 191 albatus, Myopterus daubentonii 663 Andaman Intermediate Horseshoe Arabian Trident Bat 229 Abo Bat 725, 832 Alberico’s Broad-nosed Bat 565 Bat 321 Arabian Trident Leaf-nosed Bat 229 Abo Butterfly Bat 725, 832 albericoi, Platyrrhinus 565 andamanensis, Rhinolophus 321 arabica, Asellia 229 abramus, Pipistrellus 777 albescens, Myotis 940 Andean Fruit Bat 547 arabicus, Hypsugo 810 abrasus, Cynomops 604, 640 albicollis, Megaerops 64 Andersen’s Bare-backed Fruit Bat 109 arabicus, Rousettus aegyptiacus 87 Abruzzi’s Wrinkle-lipped Bat 645 albipinnis, Taphozous longimanus 353 Andersen’s Flying Fox 158 arabium, Rhinopoma cystops 176 Abyssinian Horseshoe Bat 290 albiventer, Nyctimene 36, 118 Andersen’s Fruit-eating Bat 578 Arafura Large-footed Bat 969 Acerodon albiventris, Noctilio 405, 411 Andersen’s Leaf-nosed Bat 254 Arata Yellow-shouldered Bat 543 Sulawesi 134 albofuscus, Scotoecus 762 Andersen’s Little Fruit-eating Bat 578 Arata-Thomas Yellow-shouldered Talaud 134 alboguttata, Glauconycteris 833 Andersen’s Naked-backed Fruit Bat 109 Bat 543 Acerodon 134 albus, Diclidurus 339, 367 Andersen’s Roundleaf Bat 254 aratathomasi, Sturnira 543 Acerodon mackloti (see A. -
A Checklist of the Mammals of South-East Asia
A Checklist of the Mammals of South-east Asia A Checklist of the Mammals of South-east Asia PHOLIDOTA Pangolin (Manidae) 1 Sunda Pangolin (Manis javanica) 2 Chinese Pangolin (Manis pentadactyla) INSECTIVORA Gymnures (Erinaceidae) 3 Moonrat (Echinosorex gymnurus) 4 Short-tailed Gymnure (Hylomys suillus) 5 Chinese Gymnure (Hylomys sinensis) 6 Large-eared Gymnure (Hylomys megalotis) Moles (Talpidae) 7 Slender Shrew-mole (Uropsilus gracilis) 8 Kloss's Mole (Euroscaptor klossi) 9 Large Chinese Mole (Euroscaptor grandis) 10 Long-nosed Chinese Mole (Euroscaptor longirostris) 11 Small-toothed Mole (Euroscaptor parvidens) 12 Blyth's Mole (Parascaptor leucura) 13 Long-tailed Mole (Scaptonyx fuscicauda) Shrews (Soricidae) 14 Lesser Stripe-backed Shrew (Sorex bedfordiae) 15 Myanmar Short-tailed Shrew (Blarinella wardi) 16 Indochinese Short-tailed Shrew (Blarinella griselda) 17 Hodgson's Brown-toothed Shrew (Episoriculus caudatus) 18 Bailey's Brown-toothed Shrew (Episoriculus baileyi) 19 Long-taied Brown-toothed Shrew (Episoriculus macrurus) 20 Lowe's Brown-toothed Shrew (Chodsigoa parca) 21 Van Sung's Shrew (Chodsigoa caovansunga) 22 Mole Shrew (Anourosorex squamipes) 23 Himalayan Water Shrew (Chimarrogale himalayica) 24 Styan's Water Shrew (Chimarrogale styani) Page 1 of 17 Database: Gehan de Silva Wijeyeratne, www.jetwingeco.com A Checklist of the Mammals of South-east Asia 25 Malayan Water Shrew (Chimarrogale hantu) 26 Web-footed Water Shrew (Nectogale elegans) 27 House Shrew (Suncus murinus) 28 Pygmy White-toothed Shrew (Suncus etruscus) 29 South-east -
Phylogeography and Population Genetics of the Endemic Malagasy Bat, Macronycteris Commersoni S.S
Phylogeography and population genetics of the endemic Malagasy bat, Macronycteris commersoni s.s. (Chiroptera: Hipposideridae) Andrinajoro R. Rakotoarivelo1,2,3, Steven M. Goodman4,5, M. Corrie Schoeman6 and Sandi Willows-Munro2 1 Department of Zoology, University of Venda, Thohoyandou, Limpopo, South Africa 2 School of Life Sciences, University of Kwa-Zulu Natal, Pietermaritzburg, Kwa-Zulu Natal, South Africa 3 Natiora Ahy, Antananarivo, Madagascar 4 Field Museum of Natural History, Chicago, IL, United States of America 5 Association Vahatra, Antananarivo, Madagascar 6 School of Life Sciences, University of Kwa-Zulu Natal, Westville, Kwa-Zulu Natal, South Africa ABSTRACT Macronycteris commersoni (Hipposideridae), a bat species endemic to Madagascar, is widespread across the island and utilizes a range of habitat types including open woodland, degraded habitats, and forested areas from sea level to 1,325 m. Despite being widely distributed, there is evidence that M. commersoni exhibits morphological and bioacoustic variation across its geographical range. We investigated the fine- scale phylogeographic structure of populations in the western half of the island using extensive spatial sampling and sequence data from two mitochondrial DNA regions. Our results indicated several lineages within M. commersoni. Individuals collected from northern Madagascar formed a single monophyletic clade (clade C). A second clade (clade B) included individuals collected from the south-western portion of the island. This second clade displayed more phylogeographical partitioning with differences in mtDNA haplotypes frequency detected between populations collected in different bioclimatic regions. Lineage dispersal, genetic divergence, and timing of expansion Submitted 13 August 2018 Accepted 3 October 2018 events of M. commersoni were probably associated with Pleistocene climate fluctuations.