Northwestern Madagascar Research Articles

Schistosomiasis prevalence and low-cost diagnostics in rural Northwestern Madagascar: a pilot study

# Background Schistosomiasis is a parasitic disease that affects the health of hundreds of millions of people worldwide. Affordable diagnostic methods are necessary for developing countries to monitor schistosomiasis rates and treat infected individuals, particularly in, Madagascar, a country with the world’s fifth highest rate of schistosomiasis. # Methods We established the prevalence rate of schistosomiasis among the local population served by a clinic in rural northwestern Madagascar. Additionally, we compared different low-cost methods for schistosomiasis detection, including empirical diagnostic methods in use before this study. 500 patients were recruited in this study, of whom 380 received three tests—urine microscopy, point-of-care circulating cathodic antigen (POC-CCA) testing, and urinalysis to screen for *Schistosoma haematobium* and *Schistosoma mansoni*. # Results By combining these modalities, we recorded a schistosomiasis prevalence rate of 64.47%, which was significantly higher than the 4.84% of patients who were diagnosed and treated through previous empirical methods. # Conclusions Our results indicate that screening tests are a necessary component of schistosomiasis control programs. The clinic continued to use urinalysis strip testing and urine microscopy to detect schistosomiasis after this study and decided to discontinue the use of POC-CCA *S. mansoni* urine testing because of cost. Increased awareness of schistosomiasis resulting from this screening program led to the installation of multiple infrastructure projects in local communities to improve access to potable water, demonstrating community benefits of schistosomiasis screening that extend beyond the identification of infected individuals.

Notoraja hesperindica sp. nov., a new colorful deep-sea softnose skate (Elasmobranchii, Rajiformes, Arhynchobatidae) and first generic record from the western Indian Ocean

A new deep-sea softnose skate, Notoraja hesperindica, is described based on six specimens caught in 1230–1600 m depths off southern Mozambique and northwestern Madagascar in the western Indian Ocean. The new species is a medium-sized species of Notoraja Ishiyama, 1958 reaching a total length of at least 539 mm. It is the only Notoraja species known from the western Indian Ocean and differs from most congeners in the intense dark blue-grayish coloration. The only congeners with similarly vibrant coloration are Notoraja azurea McEachran & Last, 2008 from off southern Australia and Notoraja sapphira Seret & Last, 2009 from the Norfolk Ridge (Coral Sea and Tasman Sea), which differ from the new species in a shorter and more obtusely angled snout, a shorter head, a shorter disc, and smaller dorsolateral tail thorns, as well as more predorsal tail and total predorsal vertebrae and more pectoral-fin radials. The large dorsolateral tail thorns also distinguish the new species from most congeners except for the eastern Pacific Notoraja martinezi Concha, Ebert & Long, 2016, which has even larger, randomly patterned dorsolateral tail thorns that give the tail a barbedwire-like appearance. This congener further differs from N. hesperindica sp. nov. in the absence (vs. presence) of malar thornlets, fewer median tail thorns, a shorter snout, more tooth rows in upper and lower jaws, and more pectoral radials. A key to the species of Notoraja in the Indian Ocean is provided.

Assessment of the Impacts of Tropical Cyclone Fantala to Tanzania Coastal Line: Case Study of Zanzibar

The study investigated the impacts of tropical cyclone (TC) Fantala (11th to 27th April, 2016) to the coastal areas of Tanzania, Zanzibar in particular. Daily reanalysis data consisting of wind speed, sea level pressure (SLP), sea surface temperatures (SSTs) anomaly, and relative humidity from the National Centres for Environmental Prediction/National Center for Atmospheric Research (NCEP/NCAR) were used to analyze the variation in strength of Fantala as it was approaching the Tanzania coastal line. In addition observed rainfall from Tanzania Meteorological Authority (TMA) at Zanzibar office, Global Forecasting System (GFS) rainfall estimates and satellite images were used to visualize the impacts of tropical cyclone Fantala to Zanzibar. The results revealed that, TC Fantala was associated with deepening/decreasing in SLP (from 1012 - 1010 mb) around the north-western Madagascar and coastal Tanzania, whereas the mean SSTs was greater than 28°C and an SSTs anomaly ranged from 0 to 2.3°C. The vertical wind shear which ridged at Mozambican Channel and over north-eastern Madagascar was high enough (12 - 15 ms-1) to support the intensifying of Fantala. The thermodynamic and dynamic conditions of Fantala influenced heavy rainfall of greater than 170 mm over most stations in Zanzibar. Moreover, Fantala disrupted the temporal variability of 2016 March to May (MAM) seasonal rainfall. Besides, more than 420 people were homeless, at least 3330 houses were destroyed, and about 2 people died. As for mainland Tanzania Fantala resulted in a death of 12 people in Kilimanjaro and Arusha, more than 315 houses were washed away by flooding leading to 13,933 people being homeless. Conclusively the study calls for an extensive research work based on examining and forecasting the TCs rainfall impacts and their contribution during the two rainfall seasons of OND and MAM in Tanzania.

Observations of daytime activity in the nocturnal northern giant mouse lemur (Mirza zaza)

Abstract We provide an anecdote of daytime activity within the northern giant mouse lemur (Mirza zaza), a small threatened primate that has always been considered strictly nocturnal, in the Anabohazo forest of northwestern Madagascar. During the dry season, we witnessed two individual M. zaza travelling separately in the afternoon period of three different days. Our observations indicate that M. zaza may undertake activity that is essential for their survival within both the light and dark periods of a day-cycle, and our findings suggest that the activity cycle of this species may be highly flexible. These observations have important implications to understand the evolution of activity patterns in M. zaza, and these findings warrant future, long term studies to establish the annual activity patterns of M. zaza and to determine whether this species is potentially cathemeral and why.

Introduction to Adalatherium hui (Gondwanatheria, Mammalia) from the Late Cretaceous of Madagascar

ABSTRACT Adalatherium hui is a latest Cretaceous (Maastrichtian) gondwanatherian mammal based on only a single specimen, a virtually complete, articulated, and well-preserved skull and postcranial skeleton. The specimen is the most complete and best preserved of any mammaliaform from the Mesozoic of the southern supercontinent Gondwana. It was discovered in the Anembalemba Member of the Maevarano Formation in the Mahajanga Basin of northwestern Madagascar. The holotype specimen includes the only complete lower jaw and the only postcranial remains known for Gondwanatheria, which, other than the cranium of Vintana sertichi (also from the latest Cretaceous of Madagascar), are represented only by isolated teeth and fragmentary dentaries. Despite being represented by an immature individual, A. hui is third only to V. sertichi and Coloniatherium cilinskii as the largest Mesozoic mammaliaform (based on body fossils) from Gondwana. Here, we (1) review the paucity of mammaliamorph skull and postcranial skeletal material from the Mesozoic of Gondwana relative to the record from Laurasia; (2) review the systematic paleontology of A. hui; (3) provide an overview of the history of discovery of the holotype specimen; (4) detail the preservation of the holotype, its preparation history, and the imaging techniques used to study it; (5) provide an overview of the geological context of A. hui, which indicates that the species lived close to the end-Cretaceous extinction event in a highly seasonal, semiarid climate; and (6) estimate the body mass of A. hui in the context of other Mesozoic mammaliaforms.

Craniofacial morphology of Adalatherium hui (Mammalia, Gondwanatheria) from the Late Cretaceous of Madagascar

ABSTRACT The cranium of Adalatherium hui, as represented in the holotype and only specimen (UA 9030), is only the second known for any gondwanatherian mammal, the other being that of the sudamericid Vintana sertichi. Both Adalatherium and Vintana were recovered from the Upper Cretaceous (Maastrichtian) Maevarano Formation of northwestern Madagascar. UA 9030 is the most complete specimen of a gondwanatherian yet known and includes, in addition to the cranium, both lower jaws and a complete postcranial skeleton. Aside from Adalatherium and Vintana, gondwanatherians are otherwise represented only by isolated teeth and lower jaw fragments, belonging to eight monotypic genera from Late Cretaceous and Paleogene horizons of Madagascar, the Indian subcontinent, Africa, South America, and the Antarctic Peninsula. Although the anterior part of the cranium is very well preserved in UA 9030, the posterior part is not. Nonetheless, comparable parts of the crania of Adalatherium and Vintana indicate some level of common ancestry through possession of several synapomorphies, primarily related to the bony composition, articular relationships, and features of the snout region. Overprinted on this shared morphology are a host of autapomorphic features in each genus, some unique among mammaliaforms and some convergent upon therian mammals. The cranium of Adalatherium is compared with the crania of other mammaliamorphs, particularly those of allotherians or purported allotherians (i.e., haramiyidans, euharamiyidans, multituberculates, Cifelliodon, and Megaconus). Particular emphasis is placed on several recently described forms: the enigmatic Cifelliodon from the Early Cretaceous of Utah and several new taxa of euharamiyidans from the Late Jurassic of China.

Effects of habitat edges on vegetation structure and the vulnerable golden-brown mouse lemur (Microcebus ravelobensis) in northwestern Madagascar

BackgroundEdge effects can influence species composition and community structure as a result of changes in microenvironment and edaphic variables. We investigated effects of habitat edges on vegetation structure, abundance and body mass of one vulnerable Microcebus species in northwestern Madagascar. We trapped mouse lemurs along four 1000-m transects (total of 2424 trap nights) that ran perpendicular to the forest edge. We installed 16 pairs of 20 m2 vegetation plots along each transect and measured nine vegetation parameters. To determine the responses of the vegetation and animals to an increasing distance to the edge, we tested the fit of four alternative mathematical functions (linear, power, logistic and unimodal) to the data and derived the depth of edge influence (DEI) for all parameters.ResultsLogistic and unimodal functions best explained edge responses of vegetation parameters, and the logistic function performed best for abundance and body mass of M. ravelobensis. The DEI varied between 50 m (no. of seedlings, no. of liana, dbh of large trees [dbh ≥ 10 cm]) and 460 m (tree height of large trees) for the vegetation parameters, whereas it was 340 m for M. ravelobensis abundance and 390 m for body mass, corresponding best to the DEI of small tree [dbh < 10 cm] density (360 m). Small trees were significantly taller and the density of seedlings was higher in the interior than in the edge habitat. However, there was no significant difference in M. ravelobensis abundance and body mass between interior and edge habitats, suggesting that M. ravelobensis did not show a strong edge response in the study region. Finally, regression analyses revealed three negative (species abundance and three vegetation parameters) and two positive relationships (body mass and two vegetation parameters), suggesting an impact of vegetation structure on M. ravelobensis which may be partly independent of edge effects.ConclusionsA comparison of our results with previous findings reveals that edge effects are variable in space in a small nocturnal primate from Madagascar. Such an ecological plasticity could be extremely relevant for mitigating species responses to habitat loss and anthropogenic disturbances.

Screening for Viruses and Lemur-Associated Filara in Wild-Caught Mosquitoes From Madagascar.

Madagascar is a hotspot of biodiversity, but poverty and population growth provoke a high risk of conflict between food security and biodiversity conservation in this tropical country. Numerous vector-borne diseases, including viral infections, affect public health in Madagascar and a continuous expansion of anthropogenically used areas intensifies contact on the human-wildlife interface. However, data on human and animal pathogens in potential insect vectors is limited. Therefore, we conducted a parasitological and virological survey of 785 adult female mosquitoes between March and May 2016 at the Ankarafantsika National Park in northwestern Madagascar. Screening included Alpha-, Phlebo-, and Flaviviridae and the recently described filarial nematode species, Lemurfilaria lemuris. The predominant mosquito genus was Culex (91%), followed by Mansonia (4.1%), Anopheles (3.4%), and Aedes (0.9%). Viral screening revealed no arboviruses, but an insect-specific flavivirus in two Culex sitiens pools. No pools screened positive for the lemur-specific filarial nematode L. lemuris.

Next-generation technologies applied to age-old challenges in Madagascar

Madagascar is a biodiversity hotspot that is facing rapid rates of deforestation, habitat destruction, and poverty. Urgent action is required to document the status of biodiversity to facilitate efficacious conservation plans. With the recent advent of portable and affordable genetic technologies, it is now possible to take genomic approaches out of the lab and into the field. Mobile genetics labs can produce scientifically reproducible data under field conditions, dramatically minimizing the time between sample collection and data analysis. Here, we show “proof of concept” by deploying miniPCR bio’s miniaturized thermal cycler alongside Oxford Nanopore’s MinION DNA sequencer in Madagascar. Specifically, we deployed this technology at Anjajavy, northwestern Madagascar for rapid biodiversity assessment. We successfully extracted mouse lemur DNA, amplified and sequenced a phylogenetically informative mitochondrial gene (cytochrome-b; cytb), and thereby confirmed the presence of Danfoss’ mouse lemur (M. danfossi) within the Anjajavy Reserve. We show that a mobile genetics lab can provide expeditious results, and allow scientists to conduct genetic analyses, potentially allowing for rapid interventions under emergency conditions in situ. Additionally, mobile labs offer powerful training opportunities for in-country scientists for whom training opportunities were previously confined to ex-situ locations. By bringing genomic technologies to Madagascar and other economically challenged and biodiverse regions of the world, the next generation of scientists and conservationists can more fully implement their leadership roles. Local laboratory and training facilities are changing the polarity of research programs in Madagascar and empowering national researchers to take charge of environmental stewardship.

Two new species of Phialiphora (Spermacoceae, Rubiaceae) exemplify drought adaptations in western Madagascar

Background – Phialiphora (Spermacoceae, Rubiaceae) is a herbaceous genus restricted to unconsolidated white sands in open spaces in dry forests in northwestern Madagascar. The genus is characterized by the semi-succulent nature of its leaves and stems, its basal leaf rosette, its head-like inflorescences subtended by large leaf-like involucrate bracts and its isostylous flowers. Hitherto, only two species were described.Methods – Classical methods of herbarium taxonomy are followed.Key results – Two new Phialiphora species, P. glabrata De Block and P. valida De Block, are described, which brings the number of species in the genus to four. The two new species differ from the two previously described species by their glabrous branches, bracts, ovaries, calyces and fruits. They are easily distinguished from each other by the shape and size of the calyx lobes. A detailed description, illustration, distribution map and preliminary conservation assessment is given for the new species, which are assessed as critically endangered. We also present a key for the genus and discuss in detail the typical branching system, the capsules and the dispersal mechanism of Phialiphora. The description of these two new species exemplifies the uniqueness of the dry vegetations on unconsolidated white sands in western Madagascar, which are characterized by a high level of endemicity.

Paleopathology in a nearly complete skeleton of Majungasaurus crenatissimus (Theropoda: Abelisauridae)

Research in the Upper Cretaceous (Maastrichtian) Maevarano Formation of northwestern Madagascar has yielded numerous individuals of the abelisaurid theropod Majungasaurus crenatissimus. The number of individuals and quality of preservation allow for a detailed assessment of pathology in this species. A previous survey of Majungasaurus material collected prior to 2005 indicated assorted types of potential infectious and traumatic etiologies. Ongoing research on materials recovered since 2005 provide many additional opportunities to better characterize the anatomy and paleopathology in Majungasaurus. In particular, an almost complete skeleton of a single individual (FMNH PR 2836) preserves evidence of pre-mortem pathologies on multiple bones, including elements of the face and lower jaw, vertebrae, forelimb, ribs and gastralia. Analyses of the pathologies suggest infection of the jugal and quadratojugal, healed fractures and hypertrophic bone growth on several dorsal ribs and a gastralium, septic arthritis in the left forelimb, and evidence of healed/partially healed tooth marks on one cervical vertebra. The injuries appear to be the result of multiple non-fatal events experienced during the life of the individual, rather than a single traumatic incident. Our survey of Majungasaurus indicates most well preserved individuals lack observable pathology, whereas select individuals possess multiple pathologies, a general pattern also noted in other large, nonavian theropods. Such a pattern may result from a snowball effect wherein one injury or infection increases the likelihood of additional maladies due to functional impairment or a compromised immune system in an individual once an initial injury has occurred.

Unusual Seismic Activities in Northwestern Madagascar Using: Spot Them All

Unusual Seismic Activities in Northwestern Madagascar Using: Spot Them All

“A mere business affair”?Women in the social and commercial worlds of nineteenth-century Madagascar

ABSTRACT The port of Mahajanga in northwestern Madagascar was home to a diverse commercial community during the nineteenth century. Merchants from East Africa, India and the Arabian Peninsula drew upon extensive contacts within Madagascar to obtain commodities they would then sell to Americans and Europeans. Women living in these coastal communities, however, rarely entered into Western accounts of long-distance exchanges, despite their roles as wives, traders and community members in creating the cosmopolitan economic world of Mahajanga. While not directly participating in commerce with Americans and Europeans, women engaged in agriculture and other economic activities, in addition to forming close relationships with foreigners. To offset the Western trading documentation that ignores female contributions, this article relies upon a detailed source, the diary of British merchant John Studdy Leigh, to place women more fully into this history of exchanges. By expanding our consideration of economic contributions, we can examine how women bridged the diverse economic communities in northwestern Madagascar through their marriages with foreign merchants as well as their work in supporting commerce.

A new stump-toed frog from the transitional forests of NW Madagascar (Anura, Microhylidae, Cophylinae, Stumpffia).

A new species of the miniaturised microhylid frog genus Stumpffia, from north-western Madagascar, is described. Stumpffia froschauerisp. nov. differs from all other described Stumpffia species in colouration and morphology and is genetically divergent (≥ 7% uncorrected p-distance to all other nominal species of the genus) in a fragment of the mitochondrial 16S rRNA gene and in a segment of the nuclear Rag-1 gene. The new species is reliably known only from a few specimens collected in the Sahamalaza (and surroundings) region. Its known distribution is limited to three forest patches severely threatened by fire, drought and high levels of forest clearance, thus suggesting a classification of “Critically Endangered” according to IUCN Red List criteria.

A New Species of Croton (Euphorbiaceae) from a Madagascan Lineage Discovered in Coastal Kenya

Abstract—Croton kinondoensis, a new species from Kenya, is described and illustrated here with photographs. It is found in the sacred Kaya Kinondo Forest, one of the last remaining coastal forests patches in Kenya. Its morphology and systematic position based on ITS and trnL-F DNA sequence data clearly place it within the Adenophorus Group of Croton, a clade of ca. 15 species otherwise known only from Madagascar and the Comoros Archipelago. Its closest affinities appear to lie with Croton mayottae, from the island of Mayotte, and C. menabeensis, from northwestern Madagascar. This new species likely represents an independent dispersal of Croton from Madagascar to mainland Africa.

Relationships between climate change, human environmental impact, and megafaunal extinction inferred from a 4000-year multi-proxy record from a stalagmite from northwestern Madagascar

Stalagmite ANJ94-2 from Anjohibe Cave in northwestern Madagascar provides an exceptionally detailed and precisely dated record of changing environmental conditions that, combined with previously published data from stalagmites, wetland deposits, and archaeological sites, allows insights into past climate change, human environmental impact, and megafaunal extinction. Proxies of past conditions recovered from Stalagmite ANJ94-2 include ratios of carbon and oxygen stable isotopes (δ13C and δ18O), mineralogy (calcite and aragonite), layer-bounding surfaces, layer-specific width, and detrital material. Those proxies suggest that the natural environment changed in response to changes in rainfall at time scales of a few decades to multiple centuries; comparison with distant proxies suggests that wetter conditions in northwestern Madagascar may have been linked to cooling in the Northern Hemisphere. Carbon isotope data nonetheless suggest that the greatest environmental change in the area coincided with human introduction of swidden (tavy) agriculture about 1200 years ago, during a time not of drought but perhaps of slightly increasing wetness. The timing and extent of environmental change 1200 to 600 years ago seen in stalagmite and wetland data suggest that human modification of the landscape had a causal role in the extinction of Madagascar’s megafauna. On the other hand, the results combine with other recent research to indicate that drought was not the cause of the megafaunal extinction.

Forest edges affect ectoparasite infestation patterns of small mammalian hosts in fragmented forests in Madagascar

Habitat loss and fragmentation drive the worldwide depletion of biodiversity. Although it is known that anthropogenic disturbances severely affect host and ecosystem integrity, effects on parasites are largely understudied. This study aims to investigate if and how habitat fragmentation affects the composition of ectoparasite communities on small mammalian hosts in two networks of dry deciduous forest fragments in northwestern Madagascar. Forest sites differing in size, proportion of edge habitat and host density were studied in the Ankarafantsika National Park and in the Mariarano region. A total of 924 individuals of two mouse lemur species, Microcebus murinus (n = 200) and Microcebus ravelobensis (n = 426), and two rodent species, endemic Eliurus myoxinus (n = 114) and introduced Rattus rattus (n = 184), were captured to assess ectoparasite infestations. Ectoparasite prevalence and ectoparasite species richness were statistically related to nine ecological variables applying generalized linear mixed models. Hosts harbored ticks (Haemaphysalis microcebi), mites (Schoutedenichia microcebi, Listrophoroides spp., Laelaptidae gen. spp.) and sucking lice (Lemurpediculus spp., Polyplax sp., Hoplopleuridae gen. sp.). Parasite prevalence differed significantly between host species for all detected parasite taxa. Proximity to the forest edge led to a significant reduction in ectoparasites. Parasite-specific edge effects were observed up to a distance of 750 m from the forest edge. The obtained results imply that habitat fragmentation impacts ectoparasite communities, in particular by negatively affecting temporary parasite species. The results are best explained by an interplay of parasite life cycles, responses to changes in abiotic factors induced by edges and host-specific responses to habitat fragmentation. The negative responses of most studied ectoparasite taxa to forest edges and habitat fragmentation demonstrate their ecological vulnerability that may eventually threaten the integrity of ecosystems and potentially impact ectoparasite biodiversity worldwide.

Abiotic Factors Affecting the Cathemeral Activity of Eulemur fulvus in the Dry Deciduous Forest of North-Western Madagascar

Cathemeral primates perform significant amounts of activity during the daylight and dark portions of the 24-h cycle. Most brown lemurs have been reported to be cathemeral. A previous study reported that brown lemurs in Ankarafantsika National Park (ANP) shift their activity pattern from being cathemeral in the dry season to being diurnal in the wet season. From July 2015 to March 2016, we collected data on active behaviour of brown lemurs over 46 full-day and 33 full-night observations, distributed evenly between the dry and wet seasons. This study examined the abiotic factors that potentially allow this seasonal shift from cathemeral to diurnal activity. We analysed the effects of day length (time from sunrise to sunset), nocturnal luminosity, and climatic factors on the diurnal and nocturnal activities of the brown lemurs in ANP using generalized linear mixed models. We found that the brown lemurs were cathemeral regardless of season. Their diurnal activity increased with increasing day length and at high humidity, but decreased on days of high temperature and low lunar luminosity. Their nocturnal activity increased with decreasing day length, during bright nights, at low temperature, and at low humidity. Their diurnal and nocturnal activities decreased during heavy rainfall. These data indicate that the photoperiodic variation entrained their activity rhythm. Bright nights may provide better visibility, advantageous for the detection of predators and food, although it may increase exposure to predators. The modification of activity level with temperature variation can be a behavioural thermoregulation. The high humidity during the wet season probably reduced evaporative cooling, inducing increased body temperature and stimulating water foraging as a potential response to dehydration and active heat dissipation. Investigation on physiological responses to environmental factors is needed to fully understand cathemerality in lemurs. These findings emphasize the flexibility of diurnal and nocturnal activity level of brown lemurs according to the great seasonal variation in tropical dry forests.

Host Associations of Ectoparasites of the Gray Mouse Lemur, Microcebus murinus, in Northwestern Madagascar.

Eight species of ectoparasites were collected during 225 gray mouse lemur, Microcebus murinus (J. F. Miller), captures, in Ankarafantsika National Park, Madagascar, in 2010-2011. The ixodid tick, Haemaphysalis lemuris Hoogstraal, was the most common ectoparasite and was mostly represented by nymphs. Other ectoparasites recorded include the polyplacid sucking louse, Lemurpediculus madagascariensis Durden, Kessler, Radespiel, Zimmermann, Hasiniaina, and Zohdy; the ixodid tick, Haemaphysalis simplex Neumann; an undescribed laelapid mite in the genus Aetholaelaps; another laelapid belonging to the genus Androlaelaps; the chigger mite Schoutedenichia microcebi Stekolnikov; an undescribed species of atopomelid mite in the genus Listrophoroides; and an undescribed species of psoroptid mite in the genus Cheirogalalges. Except for the 2 species of ticks and 1 species of chigger, these ectoparasites may be host-specific to M. murinus. Total tick (H. lemuris and H. simplex) infestation was significantly greater in August than October, whereas louse (L. madagascariensis) infestation was significantly greater in October. There was no significant difference in tick infestations between male and female lemurs, but male lemurs had significantly more lice than female lemurs. Reproductive status was not a significant predictor of tick infestation in males and females.

Target-enriched DNA sequencing from historical type material enables a partial revision of the Madagascar giant stream frogs (genus Mantidactylus)

ABSTRACT The subgenus Mantidactylus is a group of frogs endemic to Madagascar, including the largest anuran species on the island. Although these frogs are common and widely distributed, their taxonomy remains unclear. Two species are currently recognised, M. grandidieri and M. guttulatus, with another available name, Rana pigra, considered to be a synonym of M. grandidieri. However, molecular studies have suggested the presence of several cryptic species within the group. Additionally, due to the lack of prominent morphological features, allocating the available names to evolutionary lineages has proven challenging. In the present study, we take a first step towards solving these problems by using fragments of the 16S mitochondrial gene and RAG1 nuclear gene from all over the range of the subgenus to describe its genetic diversity. We also use a newly designed target enrichment laboratory protocol to sequence three mitochondrial fragments from five name-holding museum specimens (as old as 120 years) in order to determine to which lineages the existing names should be applied. The study of the 16S mitochondrial gene revealed 7 geographically separated lineages, distinct enough to be considered candidate species. Out of the five museum specimens analysed, four successfully yielded DNA sequences and could be attributed to one of the aforementioned lineages. Therefore, the name Mantidactylus grandidieri should be applied to the populations from North-Eastern Madagascar, while M. guttulatus refers to populations from inland localities along the Eastern coast of the island. On the other hand, the holotype of Rana pigra did not yield enough genetic material to allow definitive identification. While our data were not sufficient to assess the status of the four lineages distributed along the Eastern coast, the populations from North-Western Madagascar were highly distinct on both the mitochondrial and nuclear markers. We thus describe them as a new species, M. radaka sp. nov. http://www.zoobank.org/urn:lsid:zoobank.org:pub:1EDDAF0D-FE37-490A-B09E-E136A0C5CB35