Can cyclone exposure explain behavioural and demographic variation among lemur species?

Madagascar has a harsh and stochastic climate because of regular natural disturbances. This history of regular cyclones has been hypothesised to have directed evolutionary changes to lemur behaviour and morphology that make them more resilient to sudden environmental change. These adaptations may include: small group sizes, high degrees of energy-conserving behaviours, generalist habitat use, small home ranges, small body size, and a limited number of frugivorous species. To date, however, no one has tested how variation in cyclone exposure across Madagascar is associated with variation in these resilience traits. In this study, we created a detailed cyclone impact map for Madagascar using Koppen-Geiger climate class, historical cyclone tracks, the Saffir Class of cyclone and hurricane intensity, and precipitation data. We also used existing literature to calculate a resilience score for 26 lemur species for which data existed on resilience traits. Our cyclone impact map was then overlaid on known geographic ranges of these species and compared to resilience score while controlling for phylogenetic non-independence and spatial autocorrelation. We found no association between cyclone impact in a lemur range and their resilience score. When assessing traits individually, however, we found that cyclone impact was positively associated with body size, suggesting that the more impacted a species is by cyclones the smaller they are. We also found cyclone impact to be negatively associated with frugivory, with species in higher impact zones eating more fruit. While unexpected, this could reflect an increased production in fruit in tree fall gaps following cyclones. While we did not find a pattern between cyclone impact and behavioural resilience in lemurs, we suggest a similar study at a global scale across all primates would allow for more taxonomic variation and reveal larger patterns key to understanding past and future vulnerability to natural disturbances in primates.

Consistency in Verreaux's sifaka home range and core area size despite seasonal variation in resource availability as assessed by Enhanced Vegetation Index (EVI).

Primates are adept at dealing with fluctuating availability of resources and display a range of responses to minimize the effects of food scarcity. An important component of primate conservation is to understand how primates adapt their foraging and ranging patterns in response to fluctuating food resources. Animals optimize resource acquisition within the home range through the selection of resource-bearing patches and choose between contrasting foraging strategies (resource-maximizing vs. area-minimizing). Our study aimed to characterize the foraging strategy of a folivorous primate, Verreaux's sifaka (Propithecus verreauxi), by evaluating whether group home range size varied between peak and lean leaf seasons within a seasonally dry tropical forest in Madagascar. We hypothesized that Verreaux's sifaka used the resource maximization strategy to select high-value resource patches so that during periods of resource depression, the home range area did not significantly change in size. We characterized resource availability (i.e., primary productivity) by season at Kirindy Mitea National Park using remotely-sensed Enhanced Vegetation Index data. We calculated group home ranges using 10 years of focal animal sampling data collected on eight groups using both 95% and 50% kernel density estimation. We used area accumulation curves to ensure each group had an adequate number of locations to reach seasonal home range asymptotes. Neither 95% home ranges nor 50% core areas differed across peak and lean leaf resource seasons, supporting the hypothesis that Verreaux's sifaka use a resource maximization strategy. With a better understanding of animal space use strategies, managers can model anticipated changes under environmental and/or anthropogenic resource depression scenarios. These findings demonstrate the value of long-term data for characterizing and understanding foraging and ranging patterns. We also illustrate the benefits of using satellite data for characterizing food resources for folivorous primates.

Dental caries in living and extinct strepsirrhines with insights into diet.

Dental caries is one of the most common diseases afflicting modern humans and occurs in both living and extinct non-human primates, as well as other mammalian species. Compared to other primates, less is known about the etiology or frequency of caries among the Strepsirrhini. Given the link between caries and diet, caries frequency may be informative about the dietary ecology of a given animal. Understanding rates of caries in wild populations is also critical to assessing dental health in captive populations. Here, we examine caries frequency in a sample of 36 extant strepsirrhine species (n = 316 individuals) using odontological collections of wild-, non-captive animals housed at the American Museum of Natural History by counting the number of specimens characterized by the disease. Additionally, in the context of studying caries lesions in strepsirrhines, case studies were also conducted to test if similar lesions were found in their fossil relatives. In particular, two fossil strepsirrhine species were analyzed: the earliest Late Eocene Karanisia clarki, and the subfossil lemur Megaladapis madagascariensis. Our results suggest that caries affects 13.92% of the extant individuals we examined. The frugivorous and folivorous taxa were characterized by the highest overall frequency of caries, whereas the insectivores, gummivores, and omnivores had much lower caries frequencies. Our results suggest that caries may be common among wild populations of strepsirrhines, and in fact is more prevalent than in many catarrhines and platyrrhines. These findings have important implications for understanding caries, diet, and health in living and fossil taxa.

An integrated passive acoustic monitoring and deep learning pipeline for black-and-white ruffed lemurs (Varecia variegata) in Ranomafana National Park, Madagascar.

The urgent need for effective wildlife monitoring solutions in the face of global biodiversity loss has resulted in the emergence of conservation technologies such as passive acoustic monitoring (PAM). While PAM has been extensively used for marine mammals, birds, and bats, its application to primates is limited. Black-and-white ruffed lemurs (Varecia variegata) are a promising species to test PAM with due to their distinctive and loud roar-shrieks. Furthermore, these lemurs are challenging to monitor via traditional methods due to their fragmented and often unpredictable distribution in Madagascar's dense eastern rainforests. Our goal in this study was to develop a machine learning pipeline for automated call detection from PAM data, compare the effectiveness of PAM versus in-person observations, and investigate diel patterns in lemur vocal behavior. We did this study at Mangevo, Ranomafana National Park by concurrently conducting focal follows and deploying autonomous recorders in May-July 2019. We used transfer learning to build a convolutional neural network (optimized for recall) that automated the detection of lemur calls (57-h runtime; recall = 0.94, F1 = 0.70). We found that PAM outperformed in-person observations, saving time, money, and labor while also providing re-analyzable data. Using PAM yielded novel insights into V. variegata diel vocal patterns; we present the first published evidence of nocturnal calling. We developed a graphic user interface and open-sourced data and code, to serve as a resource for primatologists interested in implementing PAM and machine learning. By leveraging the potential of this pipeline, we can address the urgent need for effective primate population surveys to inform conservation strategies.

Effects of forest fragmentation on the dietary ecology and activity of a nocturnal lemur community in North West Madagascar.

Deforestation and habitat fragmentation is the primary threat to primate populations. The primates that live within degraded and anthropogenically disturbed habitats typical of fragmented landscapes have to cope with lower availability of resources in comparison to primates in continuous, undisturbed forests. While some species are sensitive to forest fragmentation, some evidence exists to suggest that primates can alter their behavior and adapt to such changes, which enables their survival in suboptimal habitat. In this study, we assessed how forest fragmentation and its associated edge-effects impact the feeding ecology and activity levels of a nocturnal primate community in the Sahamalaza-Iles Radama National Park, North West Madagascar. From March 06, 2019 to May 17, 2022, we collected data on tree and invertebrate phenology at our study site, and feeding ecology and activity for 159 lemur individuals from four species. Fruit and flower availability varied significantly between continuous and fragmented forest, and between forest core and edge areas, with continuous forest exhibiting higher continuous fruit and flower availability. Lemur feeding ecology varied significantly too, as the feeding niches of all four species were significantly different between continuous and fragmented forest and between core and edge areas. However, lemur activity levels were mostly consistent among all forest areas. The results of this study suggest that nocturnal lemurs are able to adapt their dietary ecology in response to the available food sources within their habitat. Due to this flexible ecology and dietary plasticity, the lemurs do not need to significantly alter their behavior in different environments to fulfill their dietary needs. While nocturnal lemurs demonstrate adaptability and flexibility to degraded habitat, it is unclear how far this plasticity will stretch considering that Madagascar's forests are still being cleared at an alarming rate. Urgent conservation action is therefore needed to ensure the future of lemur habitat.

Activity budget and seasonal activity shifts in sympatric lemurs: Increased lean season effort in a cathemeral frugivore contrasts with energy conservation in a diurnal folivore.

One of the most fundamental aspects of a species' behavioral strategy is its activity budget; for primates this generally involves the allocation of available time among resting, feeding, traveling, and social behavior. Comparisons between species, populations, or individuals can reveal divergences in adaptive strategies and current stressors, and reflect responses to such diverse pressures as predation, thermoregulation, nutrition, and social needs. Further, variation across seasons is an important part of behavioral strategies to survive food scarcity; this can involve increasing or decreasing effort. We documented activity over the 24-h cycle for the cathemeral, frugivorous Eulemur fulvus and the diurnal, folivorous Propithecus diadema across 13-18 months at Tsinjoarivo, Madagascar. Their activity budgets were dominated by resting (E. fulvus: 74.1%; P. diadema: 85.2%), followed by feeding (15.8%, 12.4%), traveling (9.31%, 1.74%) and social activities (0.76%, 0.70%), respectively. The lower feeding and higher resting in P. diadema likely reflect slower gastrointestinal transit and higher reliance on microbial fermentation to extract energy from fibrous food. The two species showed opposite lean season strategies. E. fulvus increased activity, with more feeding but less travel time, consistent with a shift to less-profitable fruits, and some leaves and flowers, while increasing feeding effort to compensate ("energy maximizing"). P. diadema showed less variation across months, but the lean season still evoked reduced effort across the board (feeding, travel, and social behavior), consistent with a "time minimizing" strategy prioritizing energy conservation and microbe-assisted digestion. Understanding these divergent shifts is key to understanding natural behavior and the extent of behavioral flexibility under stressful conditions. Finally, the complex patterns of fruit availability (intra- and interannually) and the species' behavioral responses across months underscore the need to move beyond simplistic "lean/abundant season" and "fruit/leaf" dichotomies in understanding underlying energetic strategies, and species' vulnerability to habitat change.

Urinary C-peptide and total triiodothyronine as energetic biomarkers for studies of lemurs.

Measuring energy balance and energy metabolism can provide crucial information for understanding the ecological and behavioral drivers of an animal's energetic and physiological condition. Both urinary C-peptide (uCP) of insulin and urinary total triiodothyronine (uTT3) have been validated as noninvasive biomarkers of energy balance and metabolic activity in haplorrhine primates. This study attempts to validate uCP and uTT3 measures in strepsirrhines, a phylogenetically distinct primate clade, using the ruffed lemur (genus Varecia) as a model. We experimentally manipulated the diet of captive black-and-white (Varecia variegata) and red (Varecia rubra) ruffed lemurs at Duke Lemur Center across a 4-week period. We collected urine samples from subjects (n = 5) each day during 1 week of control diet, 2 weeks of calorie-restricted diet and 1 week of refeeding, designed to temporarily reduce energy balance and metabolism. We also tested the outcome of filter paper as a storage method by comparing to controls (frozen at -20°C) to assess its suitability for studies of wild populations. We successfully measured uCP and uTT3 levels in frozen urine samples using commercial enzyme immunoassay kits and found that both biomarkers were excreted at lower concentrations (C-peptide: 1.35 ng/mL, 54% reduction; TT3: 1.5 ng/mL, 37.5% reduction) during calorie-restricted periods compared to normal diet periods. Filter paper recovery for uCP was 19%, though values were significantly positively correlated with frozen control samples. uTT3 could not be recovered at measurable concentrations using filter paper. These methods enable noninvasive measurement of energetic conditions in wild strepsirrhines and subsequent assessment of relationships between energy balance and numerous socioecological drivers in primate populations.

The provisioned primate: patterns of obesity across lemurs, monkeys, apes and humans.

Non-human primates are potentially informative but underutilized species for investigating obesity. I examined patterns of obesity across the Primate order, calculating the ratio of body mass in captivity to that in the wild. This index, relative body mass, for n = 40 non-human primates (mean ± s.d.: females: 1.28 ± 0.30, range 0.67-1.78, males: 1.24 ± 0.28, range 0.70-1.97) overlapped with a reference value for humans (women: 1.52, men: 1.44). Among non-human primates, relative body mass was unrelated to dietary niche, and was marginally greater among female cohorts of terrestrial species. Males and females had similar relative body masses, but species with greater sexual size dimorphism (male/female mass) in wild populations had comparatively larger female body mass in captivity. Provisioned populations in wild and free-ranging settings had similar relative body mass to those in research facilities and zoos. Compared to the wild, captive diets are unlikely to be low in protein or fat, or high in carbohydrate, suggesting these macronutrients are not driving overeating in captive populations. Several primate species, including chimpanzees, a sister-species to humans, had relative body masses similar to humans. Humans are not unique in the propensity to overweight and obesity. This article is part of a discussion meeting issue 'Causes of obesity: theories, conjectures and evidence (Part II)'.

Identification of 3 novel herpesviruses in prosimians with lymphoproliferative disease.

Although many studies have characterized catarrhine and platyrrhine primate herpesviruses, little is known about herpesviruses in prosimians. We aimed to identify and characterize herpesviruses in prosimians with proliferative lymphocytic disease. DNA was extracted from tissues of 9 gray mouse lemurs (Microcebus murinus) and 3 pygmy slow lorises (Nycticebus pygmaeus) with lymphoproliferative lesions, and we performed nested PCR and sequencing for detection of herpesviruses and polyomaviruses. We identified 3 novel herpesviruses and performed phylogenetic analyses to characterize their relationship with other herpesviruses. A gray mouse lemur herpesvirus clustered with other primate herpesviruses within the subfamily Betaherpesvirinae, just basal to the genus Cytomegalovirus. The other gray mouse lemur herpesvirus and the pygmy slow loris herpesvirus clustered within the subfamily Gammaherpesvirinae, although the relationships within the subfamily were less resolved. Quantitative PCR assays were developed for the 2 new gray mouse lemur viruses, providing specific, faster, less expensive, and quantitative detection tools. Further studies are needed to elucidate the relationship between the presence of these viruses and the severity or presence of lymphoproliferative lesions in prosimians.

Mean amplitude of low frequency fluctuations measured by fMRI at 11.7 T in the aging brain of mouse lemur primate.

Non-human primates are a critical species for the identification of key biological mechanisms in normal and pathological aging. One of these primates, the mouse lemur, has been widely studied as a model of cerebral aging or Alzheimer's disease. The amplitude of low-frequency fluctuations of blood oxygenation level-dependent (BOLD) can be measured with functional MRI. Within specific frequency bands (e.g. the 0.01-0.1 Hz), these amplitudes were proposed to indirectly reflect neuronal activity as well as glucose metabolism. Here, we first created whole brain maps of the mean amplitude of low frequency fluctuations (mALFF) in young mouse lemurs (mean ± SD: 2.1 ± 0.8 years). Then, we extracted mALFF in old lemurs (mean ± SD: 8.8 ± 1.1 years) to identify age-related changes. A high level of mALFF was detected in the temporal cortex (Brodmann area 20), somatosensory areas (Brodmann area 5), insula (Brodmann areas 13-6) and the parietal cortex (Brodmann area 7) of healthy young mouse lemurs. Aging was associated with alterations of mALFF in somatosensory areas (Brodmann area 5) and the parietal cortex (Brodmann area 7).

Evaluating the impact of environmental education around Ranomafana National Park.

A total of 94% of lemur species are currently threatened with extinction and more than 17 species of giant lemur are already extinct. To help prevent the extinction of Madagascar's remaining lemurs, Dr. Patricia Wright initiated conservation programs in the Ranomafana region of southern Madagascar in the 1990s. These continued and expanded, and in 2003 were consolidated with Dr. Wright's research activities when Center ValBio ("CVB") was founded in 2003. CVB believes in the "One Health" approach in understanding the relationship between humans and the environment, and one of their core principles is that effective conservation is science-based. CVB's environmental education (EE) programs (discussed herein) operate in various primary schools surrounding Ranomafana national park (RNP). The all-Malagasy team consists of long-term conservation educators as well as young intern teachers, who together address the issues of valuing lemurs and the forests that they require to survive. In this paper, we will describe three of CVB's EE programs and evaluate their impact. The primary tool used to assess impact was an analysis of pre- and post -intervention test scores evaluated using a Kruskal-Wallis test. We show that these programs (1) are popular, (2) produce concrete outputs that can change rural villages, and (3) improve local knowledge on the importance of biodiversity and sustainable development.

Spontaneous neoplasms in captive red-bellied lemurs (Eulemur rubriventer).

Red-bellied lemurs (Eulemur rubriventer: genus Eulemur, family Lemuridae, Suborder Strepsirrhini) are non-human primates endemic to the forests of Madagascar and listed as 'vulnerable' on the IUCN Red List of Threatened Species. Currently, descriptions of neoplasia in this species are extremely scarce, with only one case of hepatocellular adenoma reported. Prosimian submissions received by the Anatomic Pathology Service at North Carolina State University from January 2010 to January 2021 were retrieved. A total of 200 cases of Strepsirrhini prosimians were identified, representing 57 (28.5%) individuals from the genus Eulemur of which seven (12.3%) cases were red-bellied lemurs. Neoplasia was identified in two of the seven (28.57%) cases. The first case was a 25-year-old, intact female with hepatocellular carcinoma and the second a 33-year-old, intact female with a cervical mass consistent with osteoblastic osteosarcoma arising from the left clavicular bone. Hepatocellular neoplasms are commonly reported in prosimians, with carcinomas more frequent than adenomas. In contrast, osteosarcoma has been rarely described in lemuriformes, with four cases reported. To our knowledge, this is the first report of osteosarcoma and hepatocellular carcinoma in a red-bellied lemur.

Seasonal changes in the parasite prevalence of a small Malagasy lemur species (Lepilemur edwardsi).

Parasitic infections can impact the fitness of individuals and can have influence on animals' population dynamics. An individuals' parasite prevalence often changes depending on external or seasonal changes, for example, rainfall and ambient temperatures, but also on internal changes, for example, changes in body condition. In this study we aimed to identify the environmental factors that may influence the intestinal parasite and ectoparasite prevalence of the folivorous Malagasy primate species, Lepilemur edwardsi, living in a seasonal dry deciduous forest. Species living in this habitat have to adapt to seasonal changes of ambient temperature, with almost no precipitation during the dry season and hence strong fluctuations of resource availability throughout the year. We sampled the feces and ectoparasites of L. edwardsi throughout the year. Intestinal parasite prevalence increased from the wet to the dry season and was highest in the late dry season, which might be due to the accompanying decrease in diet quality. Conversely, ectoparasite prevalence decreased in the dry season, presumably due to the prevailing unfavorable environmental conditions for the development of ectoparasites (i.e., mites and ticks). Paired with the higher resting metabolism and stress level of L. edwardsi during the late dry season, it seems that this species may struggle when dry seasons intensify in its habitat.

Gut Site and Gut Morphology Predict Microbiome Structure and Function in Ecologically Diverse Lemurs.

Most studies of wildlife gut microbiotas understandably rely on feces to approximate consortia along the gastrointestinal tract. We therefore compared microbiome structure and predicted metagenomic function in stomach, small intestinal, cecal, and colonic samples from 52 lemurs harvested during routine necropsies. The lemurs represent seven genera (Cheirogaleus, Daubentonia, Varecia, Hapalemur, Eulemur, Lemur, Propithecus) characterized by diverse feeding ecologies and gut morphologies. In particular, the hosts variably depend on fibrous foodstuffs and show correlative morphological complexity in their large intestines. Across host lineages, microbiome diversity, variability, membership, and function differed between the upper and lower gut, reflecting regional tradeoffs in available nutrients. These patterns related minimally to total gut length but were modulated by fermentation capacity (i.e., the ratio of small to large intestinal length). Irrespective of feeding strategy, host genera with limited fermentation capacity harbored more homogenized microbiome diversity along the gut, whereas those with expanded fermentation capacity harbored cecal and colonic microbiomes with greater diversity and abundant fermentative Ruminococcaceae taxa. While highlighting the value of curated sample repositories for retrospective comparisons, our results confirm that the need to survive on fibrous foods, either routinely or in hypervariable environments, can shape the morphological and microbial features of the lower gut.

Material and food exploration by zoo-housed animals can inform cognition and enrichment apparatus design.

To robustly study zoo animal cognition and provide effective enrichment, we must provide animals with carefully designed apparatus made from appropriate (safe, attractive, practical) materials. However, all too often, this design phase is overlooked or omitted from the literature. We evaluated how a troop of 12 ring-tailed lemurs (Lemur catta) explored a range of novel materials and whole foods during outdoor social testing. These items were not intended to test cognition or be enriching; rather we viewed them as the potential "building blocks" from which to build our future apparatus. Lemurs preferred to explore wooden surfaces, but had no preference for manipulanda made from different materials. Large amounts of metal and untreated wood should be avoided in the future; metal produced too much heat and glare, and wood was damaged by biting/chewing. Lemurs used one or two hands to explore manipulanda, and simple touching was more common than twisting or pulling. However, lemurs were most likely to explore by smell than touch or by mouth. Social testing preserved "normal" conditions for the lemurs, including natural food stealing and scrounging in high- and low-ranking individuals, respectively. Our findings culminated in the development of a static, low-level cognitive task apparatus, constructed from modular plastic units. We encourage other researchers to report how they develop cognitive and enrichment apparatuses and consider a similar preference-testing approach.

Forest access restores foraging and ranging behavior in captive sifakas.

Captive wildlife benefit from ecologically informed management strategies that promote natural behaviors. The Duke Lemur Center has pioneered husbandry programs rooted in species' ecology for a diversity of lemurs, including housing social groups in multiacre forest enclosures. We systematically document the foraging and ranging patterns of Coquerel's sifakas (Propithecus coquereli) living in these forest enclosures. Coquerel's sifakas are seasonal frugo-folivores that exhibit striking feeding flexibility in the wild. They are also one of the few members of the Indriidae family to persist in captivity. During all-day follows in the spring and summer of 2 consecutive years, we tracked the behavior of 14 sifakas in six forest enclosures. The sifakas' ranging and foraging patterns reflected those of wild sifakas in western Madagascar: On average, DLC sifakas occupied 3-day home ranges of 1.2 ha, traveled 473 m/day, and spent 26% of their time foraging for wild foodstuffs. The sifakas foraged most for young and mature leaves, fruits, nuts, and flowers from 39 plant species, especially red maple (Acer rubrum), tulip poplar (Liriodendron tulipifera), black locust (Robinia pseudoacacia), grapevine (Vitis rotundifolia), hickory (Carya spp.), and white oak (Quercus alba). Foraging patterns varied across seasons, enclosure areas, and groups, potentially reflecting differences in phenology, microhabitats, and individual preferences. While demonstrating that captive-bred primates express wild-like behaviors under ecologically relevant conditions, our results underscore the feeding flexibility of the Coquerel's sifaka. Captive wildlife exhibiting the range of species-specific behaviors are key resources for ecological research and might be best suited for future reintroductions.

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.

Nine years of the Red Book Challenge conservation education in Southern Madagascar: What we have learned.

Red Book Challenge Conservation Education Madagascar is a small-scale, arts-based, conservation education initiative founded in 2012 by two Lemur catta researchers working in the Berenty Reserve and incorporated as a nonprofit organization in 2022. The target demographic is children and adults living in Southern Madagascar. The Red Book Challenge program initially consisted of providing supplementary conservation education through classroom visits at the elementary schools in Berenty village. The school children created small conservation booklets with red covers, giving the project its name. Today, the program has expanded to include the village of Besakoa Ambany and the city of Fort Dauphin, while remaining a small-scale program. To date, the Red Book Challenge has completed seven outreach projects aimed at generating goodwill in the local community, community development, and conservation education. This retrospective analysis functions as a formative evaluation of the feasibility of future projects. This paper will focus on four projects that fall under the category of conservation education: (1) Using films to promote conservation topics (2) field trips to observe lemurs (3) a Malagasy-run conservation club, and (4) a coloring book. These projects have been funded through small grants from the International Conservation Fund of Canada, and private donations. This paper includes the Red Book Challenge's history, results from the participant surveys, and the successes and failures in each stage of our work. Six shifts in thinking occurred: (1) use of Malagasy talent to create educational conservation films (2) more formal assessment before and after field trips (3) improvement of recruitment, training and supervision of staff to keep the conservation club self-sufficient (4) enlistment of Malagasy illustrators and adaptation of environmentally friendly printing practices for the coloring books (5) disseminate study outcomes to international community though publication (6) reframe the program from spare-time volunteer work to full-time research work.

Updated lemur species ranges in Madagascar's Corridor Forestier d'Ambositra Vondrozo (COFAV).

Madagascar's high rates of endemism, paired with its escalating deforestation rates, has made it one of the most important conservation priorities on the planet. In southeastern Madagascar, the Corridor Forestier d'Ambositra Vondrozo (COFAV) is an unprotected rainforest corridor that sustains ∼15 species of lemurs, most of which are endangered. The COFAV connects many protected areas and is therefore essential for gene flow, dispersal, and the long-term sustainability of animal populations in the area. The corridor has not been surveyed extensively since the 1990s, and even so, only a fraction of the sites have been sampled multiple times. The goal of our study was to survey the COFAV, from Ranomafana National Park to the Mananara River, to provide updated species occurrences and ranges. Combining data across multiple teams using different inventorying methods, we surveyed a total area of 227 km2 throughout an eight-month period. We recorded every lemur occurrence (sighting or vocalization) and noted the species, date, time, group size, and GPS coordinates. We found 11 lemur species and one putative hybrid species. The geographic ranges for three species (Hapalemur aureus, H. griseus, Propithecus edwardsi) were larger than previously thought. The range of Varecia variegata should be shortened and adjusted accordingly, as the species appears transient (at best) in the northern parts of Ranomafana National Park and was not found south of the Ambohimahamasina/Ikongo region. This study provides updated geographic ranges for lemur species in the COFAV, important information for future censuses, species assessments, and conservation measures for future implementation.

Interspecies Papillomavirus Type Infection and a Novel Papillomavirus Type in Red Ruffed Lemurs (Varecia rubra).

The Papillomaviridae are a family of vertebrate-infecting viruses of oncogenic potential generally thought to be host species- and tissue-specific. Despite their phylogenetic relatedness to humans, there is a scarcity of data on papillomaviruses (PVs) in speciose non-human primate lineages, particularly the lemuriform primates. Varecia variegata (black-and-white ruffed lemurs) and Varecia rubra (red ruffed lemurs), two closely related species comprising the Varecia genus, are critically endangered with large global captive populations. Varecia variegata papillomavirus (VavPV) types -1 and -2, the first PVs in lemurs with a fully identified genome, were previously characterized from captive V. variegata saliva. To build upon this discovery, saliva samples were collected from captive V. rubra with the following aims: (1) to identify PVs shared between V. variegata and V. rubra and (2) to characterize novel PVs in V. rubra to better understand PV diversity in the lemuriform primates. Three complete PV genomes were determined from V. rubra samples. Two of these PV genomes share 98% L1 nucleotide identity with VavPV2, denoting interspecies infection of V. rubra by VavPV2. This work represents the first reported case of interspecies PV infection amongst the strepsirrhine primates. The third PV genome shares <68% L1 nucleotide identity with that of all PVs. Thus, it represents a new PV species and has been named Varecia rubra papillomavirus 1 (VarPV1). VavPV1, VavPV2, and VarPV1 form a new clade within the Papillomaviridae family, likely representing a novel genus. Future work diversifying sample collection (i.e., lemur host species from multiple genera, sample type, geographic location, and wild populations) is likely to uncover a world of diverse lemur PVs.