Where the small things are: Modelling edge effects on mouse lemur population density and distribution in northwestern Madagascar.

Edge effects result from the penetration to varying depths and intensities, of abiotic and biotic conditions from the surrounding non-forest matrix into the forest interior. Although 70% of the world's forests are within 1 km of a forest edge, making edge effects a dominant feature of most forest habitats, there are few empirical data on inter-site differences in edge responses in primates. We used spatially explicit capture-recapture (SECR) models to determine spatial patterns of density for two species of mouse lemurs (Microcebus murinus and Microcebus ravelobensis) in two forest landscapes in northwestern Madagascar. The goal of our study was to determine if mouse lemurs displayed spatially variable responses to edge effects. We trapped animals using Sherman live traps in the Mariarano Classified Forest (MCF) and in the Ambanjabe Forest Fragment Site (AFFS) site within Ankarafantsika National Park. We trapped 126 M. murinus and 79 M. ravelobensis at MCF and 78 M. murinus and 308 M. ravelobensis at AFFS. For M. murinus, our top model predicted a positive edge response, where density increased towards edge habitats. In M. ravelobensis, our top model predicted a negative edge response, where density was lower near the forest edges and increased towards the forest interior. At regional and landscape-specific scales, SECR models estimated different density patterns between M. murinus and M. ravelobensis as a result of variation in edge distance. The spatial variability of our results using SECR models indicate the importance of studying the population ecology of primates at varying scales that are appropriate to the processes of interest. Our results lend further support to the theory that some lemurs exhibit a form of ecological flexibility in their responses to forest loss, forest fragmentation, and associated edge effects.

Special Issue "Primate Phylogeny and Genetics".

New phylogenetic tools and population genetics methods have been developed and vastly advanced over the last decade [...].

Morphological variability or inter-observer bias? A methodological toolkit to improve data quality of multi-researcher datasets for the analysis of morphological variation.

OBJECTIVES: The investigation of morphological variation in animals is widely used in taxonomy, ecology, and evolution. Using large datasets for meta-analyses has dramatically increased, raising concerns about dataset compatibilities and biases introduced by contributions of multiple researchers. MATERIALS AND METHODS: We compiled morphological data on 13 variables for 3073 individual mouse lemurs (Cheirogaleidae, Microcebus spp.) from 25 taxa and 153 different sampling locations, measured by 48 different researchers. We introduced and applied a filtering pipeline and quantified improvements in data quality (Shapiro-Francia statistic, skewness, and excess kurtosis). The filtered dataset was then used to test for genus-wide sexual size dimorphism and the applicability of Rensch's, Allen's, and Bergmann's rules. RESULTS: Our pipeline reduced inter-observer bias (i.e., increased normality of data distributions). Inter-observer reliability of measurements was notably variable, highlighting the need to reduce data collection biases. Although subtle, we found a consistent pattern of sexual size dimorphism across Microcebus, with females being the larger (but not heavier) sex. Sexual size dimorphism was isometric, providing no support for Rensch's rule. Variations in tail length but not in ear size were consistent with the predictions of Allen's rule. Body mass and length followed a pattern contrary to predictions of Bergmann's rule. DISCUSSION: We highlighted the usefulness of large multi-researcher datasets for testing ecological hypotheses after correcting for inter-observer biases. Using genus-wide tests, we outlined generalizable patterns of morphological variability across all mouse lemurs. This new methodological toolkit aims to facilitate future large-scale morphological comparisons for a wide range of taxa and applications.

Variation in abundance and habitat use of the critically endangered Microcebus gerpi across its fragmented range.

A link between the abundance of species and their degree of ecological specialization has previously been suggested within the primate order. Many species of lemurs were only recently described and even basic ecological data are not yet available for them. We investigated the habitat use, abundance, and habitat characteristics of the critically endangered Microcebus gerpi and evaluated potential impacts of vegetation structure and human disturbances on variations in its abundance. We determined abundance by systematic nocturnal surveys along 13 transects that were also used for characterizing the vegetation structure in seven study sites that were widely distributed within its range. Although M. gerpi occurred in all studied lowland rainforest and littoral forest fragments in central eastern Madagascar and therefore has a higher ecological plasticity and wider distribution than previously thought, its actual Area of Occupancy is very small (339.78 km2 ) due to an extreme degree of habitat fragmentation throughout its range. M. gerpi occurred with a mean encounter rate of 3.04 individuals/km but abundance varied substantially between sites (0.75-4.5 individuals/km). Statistical modeling revealed that the cover of small- to medium-sized trees had a positive impact on the abundance of M. gerpi, whereas a composite disturbance score (CDS), formed on the basis of information on the prominence of fires, cattle, charcoal production and wood extraction inside and around the forest, had a negative impact on abundance. These results suggest that M. gerpi is slightly less threatened than expected because of its larger geographic range, but also that it responds negatively to human disturbances. These findings raise strong conservation concerns and question the long-term viability of the remaining small and isolated populations of this arboreal solitary forager.

Diversification processes in Gerp's mouse lemur demonstrate the importance of rivers and altitude as biogeographic barriers in Madagascar's humid rainforests.

Madagascar exhibits exceptionally high levels of biodiversity and endemism. Models to explain the diversification and distribution of species in Madagascar stress the importance of historical variability in climate conditions which may have led to the formation of geographic barriers by changing water and habitat availability. The relative importance of these models for the diversification of the various forest-adapted taxa of Madagascar has yet to be understood. Here, we reconstructed the phylogeographic history of Gerp's mouse lemur (Microcebus gerpi) to identify relevant mechanisms and drivers of diversification in Madagascar's humid rainforests. We used restriction site associated DNA (RAD) markers and applied population genomic and coalescent-based techniques to estimate genetic diversity, population structure, gene flow and divergence times among M. gerpi populations and its two sister species M. jollyae and M. marohita. Genomic results were complemented with ecological niche models to better understand the relative barrier function of rivers and altitude. We show that M. gerpi diversified during the late Pleistocene. The inferred ecological niche, patterns of gene flow and genetic differentiation in M. gerpi suggest that the potential for rivers to act as biogeographic barriers depended on both size and elevation of headwaters. Populations on opposite sides of the largest river in the area with headwaters that extend far into the highlands show particularly high genetic differentiation, whereas rivers with lower elevation headwaters have weaker barrier functions, indicated by higher migration rates and admixture. We conclude that M. gerpi likely diversified through repeated cycles of dispersal punctuated by isolation to refugia as a result of paleoclimatic fluctuations during the Pleistocene. We argue that this diversification scenario serves as a model of diversification for other rainforest taxa that are similarly limited by geographic factors. In addition, we highlight conservation implications for this critically endangered species, which faces extreme habitat loss and fragmentation.

Effects of Forest Fragmentation on Connectivity and Genetic Diversity in an Endemic and an Invasive Rodent in Northwestern Madagascar.

Habitat loss and fragmentation are of concern to conservation biologists worldwide. However, not all organisms are affected equally by these processes; thus, it is important to study the effects of living in fragmented habitats on species that differ in lifestyle and habitat requirements. In this study, we examined the dispersal and connectivity patterns of rodents, one endemic (Eliurus myoxinus) and one invasive (Rattus rattus), in two landscapes containing forest fragments and adjacent continuous forest patches in northwestern Madagascar. We generated genetic (RADseq) data for 66 E. myoxinus and 81 R. rattus individuals to evaluate differences in genetic diversity as well as inbreeding and connectivity in two landscapes. We found higher levels of inbreeding and lower levels of genetic diversity in E. myoxinus compared with R. rattus. We observed related dyads both within and between habitat patches and positive spatial autocorrelation at lower distance classes for both species, with a stronger pattern of spatial autocorrelation in R. rattus. Across each site, we identified contrasting migration rates for each species, but these did not correspond to habitat-matrix dichotomies. The relatively low genetic diversity in the endemic E. myoxinus suggests ecological constraints that require further investigation.

Host-related and environmental factors influence long-term ectoparasite infestation dynamics of mouse lemurs in northwestern Madagascar.

Parasite infestations depend on multiple host-related and environmental factors. In the case of ectoparasites, which are exposed to the environment beyond the host, an impact of climate, expressed by seasonal or yearly variations, can be expected. However, long-term dynamics of ectoparasite infestations are rarely studied in nonhuman primates. We investigated the yearly variations in ectoparasite infestations of two small primates, the gray (Microcebus murinus) and the golden-brown (Microcebus ravelobensis) mouse lemur. For a more comprehensive evaluation, we also analyzed the potential effects of yearly and monthly climatic variation (temperature, rainfall) in addition to habitat, host sex, age, species, and body mass, on ectoparasite infestation. Individuals of both host species were sampled in two study sites within the Ankarafantsika National Park in northwestern Madagascar during several months (March-November) and across 4 years (2010, 2011, 2015, 2016). Our results show significant monthly and yearly variations in the infestation rates of three native ectoparasite taxa (Haemaphysalis spp. ticks, Schoutedenichia microcebi chigger mites, Lemurpediculus spp. sucking lice) and in ectoparasite species richness in both mouse lemur species. In addition, significant impacts of several host-related (species, sex, body mass) and environmental factors (habitat, temperature, rainfall) were found, but with differences in relevance for the different parasite taxa and partly deviating in their direction. Although some differences could be attributed to either permanent or temporary presence of the parasites on the host or to ecological differences between the host species, the lack of specific knowledge regarding the life cycle and microhabitat requirements of each parasite taxon precludes a complete understanding of the factors that determine their infestation dynamics. This study demonstrates the presence of yearly and monthly dynamics in lemur-parasite interactions in tropical, seasonal, dry deciduous forests in Madagascar, which call out for broad ecological long-term studies focusing both on primate hosts and their parasites.

Habitat and ecological niche characteristics of the elusive Hairy-eared Dwarf Lemur (Allocebus trichotis) with updated occurrence and geographic range data.

Ecological niche modeling (ENM) is particularly useful in the study of cryptic and elusive species that are only rarely seen, difficult to study in the wild, and simultaneously threatened by a multitude of anthropogenic factors. We collected presence records one such example, the understudied Hairy-eared Dwarf Lemur (Allocebus trichotis), from the literature and supplemented these with observations from surveys in previously unstudied areas to model its fundamental and realized niche throughout Madagascar. We further evaluated habitat characteristics at each sighting location to judge its conservation status and its adaptability toward habitat degradation. We found 31 presence localities in the literature and directly observed A. trichotis a further 23 times. According to the ENMs, precipitation of the driest quarter and maximum temperature of the warmest month mostly influenced its distribution, restricting it to the higher altitudes of the northeastern humid forests of Madagascar. Its fundamental niche, extent of occurrence, and area of occupancy were estimated as 65,819, 53,054, and 16,967 km² respectively. The latter, however, declined by 29.2% since 1990. On a habitat level, it was most often observed in primary forest habitats of different degradation stages (85.7%, n = 21). The fundamental niche of A. trichotis is most likely restricted by climatic factors. Although it can be found in moderately degraded forests, it is known to depend on the availability of tree holes and therefore old-growth forest structures. Ongoing deforestation and habitat fragmentation are likely to be the major threats to the survival of this species. Our ENMs provide evidence on where to intensify survey efforts on the elusive Hairy-eared Dwarf Lemurs of Madagascar.

Molecular phylogenetics of the sucking louse genus Lemurpediculus (Insecta: Phthiraptera), ectoparasites of lemurs, with descriptions of three new species.

Sucking lice live in intimate association with their hosts and often display a high degree of host specificity. The present study investigated sucking lice of the genus Lemurpediculus from six mouse lemur (Microcebus) and two dwarf lemur (Cheirogaleus) species endemic to the island of Madagascar, considered a biodiversity hotspot. Louse phylogenetic trees were created based on cytochrome C oxidase subunit I (COI), elongation factor 1α (EF1α) and internal transcribed spacer 1 (ITS1) sequences. While clustering according to host species was generally observed for COI and ITS1, suggesting high host specificity of the examined lice, EF1α sequences alone did not distinguish between lice of different Microcebus species, possibly due to rather recent divergence. As bootstrap support for basal tree structure was rather low, further data are necessary to resolve the evolutionary history of louse-mouse lemur associations. Three new species of sucking lice are described: Lemurpediculus zimmermanni sp. Nov. From Microcebus ravelobensis, Lemurpediculus gerpi sp.nov. from Microcebus gerpi, and Lemurpediculus tsimanampesotsae sp. nov. from Microcebus griseorufus. These new species are compared with all known congeneric species and identifying features are illustrated for all known species of Lemurpediculus.

Long-term host-pathogen evolution of endogenous beta- and gammaretroviruses in mouse lemurs with little evidence of recent retroviral introgression.

Madagascar's flora and fauna have evolved in relative isolation since the island split from the African and Indian continents. When the last common ancestors of lemurs left Africa between 40 and 70 million years ago, they carried a subset of the viral diversity of the mainland population within them, which continued to evolve throughout the lemur radiation. Relative to other primate radiations, we know very little about the past or present viral diversity of lemurs, particularly mouse lemurs. Using high-throughput sequencing, we identified two gammaretroviruses and three betaretroviruses in the genomes of four species of wild mouse lemurs. The two gammaretroviruses and two betaretroviruses have not previously been described. One betaretrovirus was previously identified. All identified viruses are present in both Lorisiformes and Lemuriformes but absent from haplorrhine primates. The estimated ages of these viruses are consistent with the estimated divergence dates of the host lineages, suggesting they colonized the lemur genome after the Haplorrhine-Strepsirrhine split, but before the Lorisiformes-Lemuriformes split and before the colonization of Madagascar. The viral phylogenies connect multiple lineages of retroviruses from non-lemur and non-Madagascar-native species, suggesting substantial cross-species transmission occurred deep in the primate clade prior to its geographic dispersal. These phylogenies provide novel insights into known retroviral clades. They suggest that the origin of gammaretroviruses in rodents or bats may be premature and that the Jaagsiekte sheep virus clade may be older and more broadly distributed among mammals than previously thought.

Filarial infections in lemurs: Evidence for a wide geographical distribution and low host specificity among lemur species.

The relevance of emerging infectious diseases continues to grow worldwide as human activities increasingly extend into formerly remote natural areas. This is particularly noticeable on the island of Madagascar. As closest relatives to humans on the island, lemurs are of particular relevance as a potential origin of zoonotic pathogen spillover. Knowledge of pathogens circulating in lemur populations is, however, very poor. Particularly little is known about lemur hemoparasites. To infer host range, ecological and geographic spread of the recently described hemoparasitic nematode Lemurfilaria lemuris in northwestern Madagascar, a total of 942 individuals of two mouse lemur species (Microcebus murinus [n = 207] and Microcebus ravelobensis [n = 433]) and two rodent species (the endemic Eliurus myoxinus [n = 118] and the invasive Rattus rattus [n = 184]) were captured in two fragmented forest landscapes (Ankarafantsika National Park and Mariarano Classified Forest) in northwestern Madagascar for blood sample examination. No protozoan hemoparasites were detected by microscopic blood smear screening. Microfilaria were present in 1.0% (2/207) of M. murinus and 2.1% (9/433) of M. ravelobensis blood samples but not in rodent samples. Internal transcribed spacer 1 (ITS-1) sequences were identical to an unnamed Onchocercidae species previously described to infect a larger lemur species, Propithecus verreauxi, about 650 km further south. In contrast to expectations, L. lemuris was not detected. The finding of a pathogen in a distantly related host species, at a considerable geographic distance from the location of its original detection, instead of a microfilaria species previously described for one of the studied host species in the same region, illustrates our low level of knowledge of lemur hemoparasites, their host ranges, distribution, modes of transmission, and their zoonotic potential. Our findings shall stimulate new research that will be of relevance for both conservation medicine and human epidemiology.

Retraction: Teixeira et al. RADseq Data Suggest Occasional Hybridization between Microcebus murinus and M. ravelobensis in Northwestern Madagascar. Genes 2022, 13, 913.

The published article [...].

Population genomic structure in Goodman's mouse lemur reveals long-standing separation of Madagascar's Central Highlands and eastern rainforests.

Madagascar's Central Highlands are largely composed of grasslands, interspersed with patches of forest. The historical perspective was that Madagascar's grasslands had anthropogenic origins, but emerging evidence suggests that grasslands were a component of the pre-human Central Highlands vegetation. Consequently, there is now vigorous debate regarding the extent to which these grasslands have expanded due to anthropogenic pressures. Here, we shed light on the temporal dynamics of Madagascar's vegetative composition by conducting a population genomic investigation of Goodman's mouse lemur (Microcebus lehilahytsara; Cheirogaleidae). These small-bodied primates occur both in Madagascar's eastern rainforests and in the Central Highlands, making them a valuable indicator species. Population divergences among forest-dwelling mammals will reflect changes to their habitat, including fragmentation, whereas patterns of post-divergence gene flow can reveal formerly wooded migration corridors. To explore these patterns, we used RADseq data to infer population genetic structure, demographic models of post-divergence gene flow, and population size change through time. The results offer evidence that open habitats are an ancient component of the Central Highlands, and that widespread forest fragmentation occurred naturally during a period of decreased precipitation near the last glacial maximum. Models of gene flow suggest that migration across the Central Highlands has been possible from the Pleistocene through the recent Holocene via riparian corridors. Though our findings support the hypothesis that Central Highland grasslands predate human arrival, we also find evidence for human-mediated population declines. This highlights the extent to which species imminently threatened by human-mediated deforestation may already be vulnerable from paleoclimatic conditions.

RADseq Data Suggest Occasional Hybridization between Microcebus murinus and M. ravelobensis in Northwestern Madagascar.

The occurrence of natural hybridization has been reported in a wide range of organisms, including primates. The present study focuses on the endemic lemurs of Madagascar, primates for which only a few species occur in sympatry or parapatry with congeners, thereby creating limited opportunity for natural hybridization. This study examines RADseq data from 480 individuals to investigate whether the recent expansion of Microcebus murinus towards the northwest and subsequent secondary contact with Microcebus ravelobensis has resulted in the occurrence of hybridization between the two species. Admixture analysis identified one individual with 26% of nuclear admixture, which may correspond to an F2- or F3-hybrid. A composite-likelihood approach was subsequently used to test the fit of alternative phylogeographic scenarios to the genomic data and to date introgression. The simulations yielded support for low levels of gene flow (2Nm0 = 0.063) between the two species starting before the Last Glacial Maximum (between 54 and 142 kyr). Since M. murinus most likely colonized northwestern Madagascar during the Late Pleistocene, the rather recent secondary contact with M. ravelobensis has likely created the opportunity for occasional hybridization. Although reproductive isolation between these distantly related congeners is not complete, it is effective in maintaining species boundaries.

AA-amyloidosis in captive northern tree shrews (Tupaia belangeri).

A high prevalence of AA-amyloidosis was identified in a breeding colony of northern tree shrews (Tupaia belangeri) in a retrospective analysis, with amyloid deposits in different organs being found in 26/36 individuals (72%). Amyloid deposits, confirmed by Congo red staining, were detected in kidneys, intestines, skin, and lymph nodes, characteristic of systemic amyloidosis. Immunohistochemically, the deposited amyloid was intensely positive with anti-AA-antibody (clone mc4), suggesting AA-amyloidosis. The kidneys were predominantly affected (80%), where amyloid deposits ranged from mild to severe and was predominantly located in the renal medulla. In addition, many kidneys contained numerous cysts with atrophy of the renal parenchyma. There was no significant association between concurrent neoplastic or inflammatory processes and amyloidosis. The lack of distinctive predisposing factors suggests a general susceptibility of captive T. belangeri to develop amyloidosis. Clinical and laboratory findings of a female individual with pronounced kidney alterations were indicative of renal failure. The observed tissue tropism with pronounced kidney alterations, corresponding renal dysfunction, and an overall high prevalence suggests amyloidosis as an important disease in captive tree shrews.

Impact of model assumptions on demographic inferences: the case study of two sympatric mouse lemurs in northwestern Madagascar.

BACKGROUND: Quaternary climate fluctuations have been acknowledged as major drivers of the geographical distribution of the extraordinary biodiversity observed in tropical biomes, including Madagascar. The main existing framework for Pleistocene Malagasy diversification assumes that forest cover was strongly shaped by warmer Interglacials (leading to forest expansion) and by cooler and arid glacials (leading to forest contraction), but predictions derived from this scenario for forest-dwelling animals have rarely been tested with genomic datasets. RESULTS: We generated genomic data and applied three complementary demographic approaches (Stairway Plot, PSMC and IICR-simulations) to infer population size and connectivity changes for two forest-dependent primate species (Microcebus murinus and M. ravelobensis) in northwestern Madagascar. The analyses suggested major demographic changes in both species that could be interpreted in two ways, depending on underlying model assumptions (i.e., panmixia or population structure). Under panmixia, the two species exhibited larger population sizes across the Last Glacial Maximum (LGM) and towards the African Humid Period (AHP). This peak was followed by a population decline in M. ravelobensis until the present, while M. murinus may have experienced a second population expansion that was followed by a sharp decline starting 3000 years ago. In contrast, simulations under population structure suggested decreasing population connectivity between the Last Interglacial and the LGM for both species, but increased connectivity during the AHP exclusively for M. murinus. CONCLUSION: Our study shows that closely related species may differ in their responses to climatic events. Assuming that Pleistocene climatic conditions in the lowlands were similar to those in the Malagasy highlands, some demographic dynamics would be better explained by changes in population connectivity than in population size. However, changes in connectivity alone cannot be easily reconciled with a founder effect that was shown for M. murinus during its colonization of the northwestern Madagascar in the late Pleistocene. To decide between the two alternative models, more knowledge about historic forest dynamics in lowland habitats is necessary. Altogether, our study stresses that demographic inferences strongly depend on the underlying model assumptions. Final conclusions should therefore be based on a comparative evaluation of multiple approaches.

Past environmental changes affected lemur population dynamics prior to human impact in Madagascar.

Quaternary climatic changes have been invoked as important drivers of species diversification worldwide. However, the impact of such changes on vegetation and animal population dynamics in tropical regions remains debated. To overcome this uncertainty, we integrated high-resolution paleoenvironmental reconstructions from a sedimentary record covering the past 25,000 years with demographic inferences of a forest-dwelling primate species (Microcebus arnholdi), in northern Madagascar. Result comparisons suggest that climate changes through the African Humid Period (15.2 - 5.5 kyr) strongly affected the demographic dynamics of M. arnholdi. We further inferred a population decline in the last millennium which was likely shaped by the combination of climatic and anthropogenic impacts. Our findings demonstrate that population fluctuations in Malagasy wildlife were substantial prior to a significant human impact. This provides a critical knowledge of climatically driven, environmental and ecological changes in the past, which is essential to better understand the dynamics and resilience of current biodiversity.

Habitat fragmentation and vegetation structure impact gastrointestinal parasites of small mammalian hosts in Madagascar.

Deleterious effects of habitat loss and fragmentation on biodiversity have been demonstrated in numerous taxa. Although parasites represent a large part of worldwide biodiversity, they are mostly neglected in this context. We investigated the effects of various anthropogenic environmental changes on gastrointestinal parasite infections in four small mammal hosts inhabiting two landscapes of fragmented dry forest in northwestern Madagascar. Coproscopical examinations were performed on 1,418 fecal samples from 903 individuals of two mouse lemur species, Microcebus murinus (n = 199) and M. ravelobensis (n = 421), and two rodent species, the native Eliurus myoxinus (n = 102) and the invasive Rattus rattus (n = 181). Overall, sixteen parasite morphotypes were detected and significant prevalence differences between host species regarding the most common five parasites may be explained by parasite-host specificity or host behavior, diet, and socioecology. Ten host- and habitat-related ecological variables were evaluated by generalized linear mixed modeling for significant impacts on the prevalence of the most abundant gastrointestinal parasites and on gastrointestinal parasite species richness (GPSR). Forest maturation affected homoxenous parasites (direct life cycle) by increasing Lemuricola, but decreasing Enterobiinae gen. sp. prevalence, while habitat fragmentation and vegetation clearance negatively affected the prevalence of parasites with heterogenic environment (i.e., Strongyloides spp.) or heteroxenous (indirect cycle with intermediate host) cycles, and consequently reduced GPSR. Forest edges and forest degradation likely change abiotic conditions which may reduce habitat suitability for soil-transmitted helminths or required intermediate hosts. The fragility of complex parasite life cycles suggests understudied and potentially severe effects of decreasing habitat quality by fragmentation and degradation on hidden ecological networks that involve parasites. Since parasites can provide indispensable ecological services and ensure stability of ecosystems by modulating animal population dynamics and nutrient pathways, our study underlines the importance of habitat quality and integrity as key aspects of conservation.

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.

Cryptic Patterns of Speciation in Cryptic Primates: Microendemic Mouse Lemurs and the Multispecies Coalescent.

Mouse lemurs (Microcebus) are a radiation of morphologically cryptic primates distributed throughout Madagascar for which the number of recognized species has exploded in the past two decades. This taxonomic revision has prompted understandable concern that there has been substantial oversplitting in the mouse lemur clade. Here, we investigate mouse lemur diversity in a region in northeastern Madagascar with high levels of microendemism and predicted habitat loss. We analyzed RADseq data with multispecies coalescent (MSC) species delimitation methods for two pairs of sister lineages that include three named species and an undescribed lineage previously identified to have divergent mtDNA. Marked differences in effective population sizes, levels of gene flow, patterns of isolation-by-distance, and species delimitation results were found among the two pairs of lineages. Whereas all tests support the recognition of the presently undescribed lineage as a separate species, the species-level distinction of two previously described species, M. mittermeieri and M. lehilahytsara is not supported-a result that is particularly striking when using the genealogical discordance index (gdi). Nonsister lineages occur sympatrically in two of the localities sampled for this study, despite an estimated divergence time of less than 1 Ma. This suggests rapid evolution of reproductive isolation in the focal lineages and in the mouse lemur clade generally. The divergence time estimates reported here are based on the MSC calibrated with pedigree-based mutation rates and are considerably more recent than previously published fossil-calibrated relaxed-clock estimates. We discuss the possible explanations for this discrepancy, noting that there are theoretical justifications for preferring the MSC estimates in this case. [Cryptic species; effective population size; microendemism; multispecies coalescent; speciation; species delimitation.].