The East Indies reptile tick Amblyomma helvolum Koch, 1844 (Acari: Ixodidae), taxonomy, biology and new host records, including the first record of human infestation.

Amblyomma helvolum is a widespread, generalist ectoparasite of reptiles in the oriental region, and has the potential to become highly invasive should it be inadvertently introduced outside its native range through the exotic pet trade. All life stages of A. helvolum are re-characterised morphologically and the first examples of nanism (dwarfism) and gynandromorphy (male and female tissue in one animal) for the species are described. Eighteen new hosts records are presented for A. helvolum, including the first case of human infestation. The taxonomy, distribution, ecology, phenology, disease associations, and invasion biology of the species are also discussed.

How do King Cobras move across a major highway? Unintentional wildlife crossing structures may facilitate movement.

Global road networks continue to expand, and the wildlife responses to these landscape-level changes need to be understood to advise long-term management decisions. Roads have high mortality risk to snakes because snakes typically move slowly and can be intentionally targeted by drivers.We investigated how radio-tracked King Cobras (Ophiophagus hannah) traverse a major highway in northeast Thailand, and if reproductive cycles were associated with road hazards.We surveyed a 15.3 km stretch of Highway 304 to determine if there were any locations where snakes could safely move across the road (e.g., culverts and bridges). We used recurse analyses to detect possible road-crossing events, and used dynamic Brownian Bridge Movement Models (dBBMMs) to show movement pathways association with possible unintentional crossing structures. We further used Integrated Step Selection Functions (ISSF) to assess seasonal differences in avoidance of major roads for adult King Cobras in relation to reproductive state.We discovered 32 unintentional wildlife crossing locations capable of facilitating King Cobra movement across the highway. While our dBBMMs broadly revealed underpasses as possible crossing points, they failed to identify specific underpasses used by telemetered individuals; however, the tracking locations pre- and post-crossing and photographs provided strong evidence of underpass use. Our ISSF suggested a lower avoidance of roads during the breeding season, although the results were inconclusive. With the high volume of traffic, large size of King Cobras, and a 98.8% success rate of crossing the road in our study (nine individuals: 84 crossing attempts with one fatality), we strongly suspect that individuals are using the unintentional crossing structures to safely traverse the road.Further research is needed to determine the extent of wildlife underpass use at our study site. We propose that more consistent integration of drainage culverts and bridges could help mitigate the impacts of roads on some terrestrial wildlife.

Native Burmese pythons exhibit site fidelity and preference for aquatic habitats in an agricultural mosaic.

Animal movement and resource use are tightly linked. Investigating these links to understand how animals use space and select habitats is especially relevant in areas affected by habitat fragmentation and agricultural conversion. We set out to explore the space use and habitat selection of Burmese pythons (Python bivittatus) in a heterogenous, agricultural landscape within the Sakaerat Biosphere Reserve, northeast Thailand. We used VHF telemetry to record the daily locations of seven Burmese pythons and created dynamic Brownian Bridge Movement Models to produce occurrence distributions and model movement extent and temporal patterns. To explore relationships between movement and habitat selection we used integrated step selection functions at both the individual and population level. Burmese pythons had a mean 99% occurrence distribution contour of 98.97 ha (range 9.05-285.56 ha). Furthermore, our results indicated that Burmese pythons had low mean individual motion variance, indicating infrequent moves and long periods at a single location. In general, Burmese pythons restricted movement and selected aquatic habitats but did not avoid potentially dangerous land use types like human settlements. Although our sample is small, we suggest that Burmese pythons are capitalizing on human disturbed landscapes.

Can post-capture photographic identification as a wildlife marking technique be undermined by observer error? A case study using King Cobras in northeast Thailand.

Identifying individuals with natural markings is increasing in popularity to non-invasively support population studies. However, applying natural variation among individuals requires careful evaluation among target species, snakes for example have little validation of such methods. Here we introduce a mark-free identification method for King Cobras (Ophiophagus hannah) from the Sakaerat Biosphere Reserve, in northeast Thailand using both subcaudal scale pholidosis (scale arrangement and number) and unique ventral body markings to distinguish individuals. This project aims to evaluate the impact of observer error on individual identification. Observers of varying expertise, will distinguish between King Cobra individuals using identifying photographs from a previous study. We will ask randomly assigned observers to distinguish individuals via: 1) subcaudal pholidosis, 2) ventral body markings, and 3) combination of both measures. Using Bayesian logistic regression, we will assess the probability observers correctly distinguish individuals. Based on exploratory observations, we hypothesise that there will be a high probability of correct identifications using subcaudal pholidosis and ventral body markings. We aim to stimulate other studies implementing identification techniques for scrutinous assessment of such methods, in order to avoid subsequent errors during long-term population studies.

No room to roam: King Cobras reduce movement in agriculture.

BACKGROUND: Studying animal movement provides insights into how animals react to land-use changes. As agriculture expands, we can use animal movement to examine how animals change their behaviour in response. Recent reviews show a tendency for mammalian species to reduce movements in response to increased human landscape modification, but reptile movements have not been as extensively studied. METHODS: We examined movements of a large reptilian predator, the King Cobra (Ophiophagus hannah), in Northeast Thailand. We used a consistent regime of radio telemetry tracking to document movements across protected forest and adjacent agricultural areas. Using dynamic Brownian Bridge Movement Model derived motion variance, Integrated Step-Selection Functions, and metrics of site reuse, we examined how King Cobra movements changed in agricultural areas. RESULTS: Motion variance values indicated that King Cobra movements increased in forested areas and tended to decrease in agricultural areas. Our Integrated Step-Selection Functions revealed that when moving in agricultural areas King Cobras restricted their movements to remain within vegetated semi-natural areas, often located along the banks of irrigation canals. Site reuse metrics of residency time and number of revisits appeared unaffected by distance to landscape features (forests, semi-natural areas, settlements, water bodies, and roads). Neither motion variance nor reuse metrics were consistently affected by the presence of threatening landscape features (e.g. roads, human settlements), suggesting that King Cobras will remain in close proximity to threats, provided habitat patches are available. CONCLUSIONS: Although King Cobras displayed individual heterogeneity in their response to agricultural landscapes, the overall trend suggested reduced movements when faced with fragmented habitat patches embedded in an otherwise inhospitable land-use matrix. Movement reductions are consistent with findings for mammals and forest specialist species.

The pollination system of the widely distributed mammal-pollinated Mucuna macrocarpa (Fabaceae) in the tropics.

Although the pollinators of some plant species differ across regions, only a few mammal-pollinated plant species have regional pollinator differences in Asia. Mucuna macrocarpa (Fabaceae) is pollinated by squirrels, flying foxes, and macaques in subtropical and temperate islands. In this study, the pollination system of M. macrocarpa was identified in tropical Asia, where the genus originally diversified. This species requires "explosive opening" of the flower, where the wing petals must be pressed down and the banner petal pushed upward to fully expose the stamens and pistil. A bagging experiment showed that fruits did not develop in inflorescences (n = 66) with unopened flowers, whereas fruits developed in 68.7% of inflorescences (n = 131) with opened flowers. This indicated that the explosive opening is needed for the species to reproduce. Four potential pollinator mammals were identified by a video camera-trap survey, and 78.3% and 60.1% of monitored inflorescences (n = 138) were opened by gray-bellied squirrels (Callosciurus caniceps) and Finlayson's squirrels (C. finlaysonii), respectively, even though more than 10 mammal species visited flowers. Nectar was surrounded by the calyx, and the volume and sugar concentration of secreted nectar did not change during the day. This nectar secretion pattern is similar to those reported by previous studies in other regions. These results showed that the main pollinators of M. macrocarpa in the tropics are squirrels. However, the species' nectar secretion pattern is not specifically adapted to this particular pollinator. Pollinators of M. macrocarpa differ throughout the distribution range based on the fauna present, but there might not have been no distinctive changes in the attractive traits that accompanied these changes in pollinators.

Is Leopoldamys neilli (Rodentia, Muridae) a synonym of Leopoldamys herberti? A reply to Balakirev et al. (2013).

Recently, Balakirev et al. (2013) presented a taxonomic revision of the genus Leopoldamys based on phylogenetic analyses. They identified five main Leopoldamys genetic lineages and suggested to rename several of them. According to these authors, the genetic lineage previously thought to belong to L. edwardsi (lineage L1) should be assigned to L. revertens while L. neilli (lineage L2) should be considered as a junior synonym of L. herberti. Using molecular and morphological data from a large sampling of Leopoldamys specimens, the aim of the present study was to investigate the taxonomic status of L. herberti and L. neilli. This study reveals that, contrary to Balakirev et al.'s statement, both genetic lineages L1 and L2 occur in Nakhon Ratchasima Province, close to the type locality of L. herberti. We also show that the external measurements and color pattern of L. herberti are highly similar to those of L1 specimens but are not consistent with the morphology of L2 specimens. Therefore these results strongly suggest that L. herberti should be assigned to the genetic lineage L1. Consequently L. neilli should not be considered as a junior synonym of L. herberti and this study confirms that the appropriate name of the genetic lineage L2 is L. neilli. Moreover, as our results show that L. herberti should be assigned to the lineage L1, this name has nomenclatural priority over L. revertens, the species name suggested by Balakirev et al. (2013) for this lineage.

Combined mitochondrial and nuclear markers revealed a deep vicariant history for Leopoldamys neilli, a cave-dwelling rodent of Thailand.

BACKGROUND: Historical biogeography and evolutionary processes of cave taxa have been widely studied in temperate regions. However, Southeast Asian cave ecosystems remain largely unexplored despite their high scientific interest. Here we studied the phylogeography of Leopoldamys neilli, a cave-dwelling murine rodent living in limestone karsts of Thailand, and compared the molecular signature of mitochondrial and nuclear markers. METHODOLOGY/PRINCIPAL FINDINGS: We used a large sampling (n = 225) from 28 localities in Thailand and a combination of mitochondrial and nuclear markers with various evolutionary rates (two intronic regions and 12 microsatellites). The evolutionary history of L. neilli and the relative role of vicariance and dispersal were investigated using ancestral range reconstruction analysis and Approximate Bayesian computation (ABC). Both mitochondrial and nuclear markers support a large-scale population structure of four main groups (west, centre, north and northeast) and a strong finer structure within each of these groups. A deep genealogical divergence among geographically close lineages is observed and denotes a high population fragmentation. Our findings suggest that the current phylogeographic pattern of this species results from the fragmentation of a widespread ancestral population and that vicariance has played a significant role in the evolutionary history of L. neilli. These deep vicariant events that occurred during Plio-Pleistocene are related to the formation of the Central Plain of Thailand. Consequently, the western, central, northern and northeastern groups of populations were historically isolated and should be considered as four distinct Evolutionarily Significant Units (ESUs). CONCLUSIONS/SIGNIFICANCE: Our study confirms the benefit of using several independent genetic markers to obtain a comprehensive and reliable picture of L. neilli evolutionary history at different levels of resolution. The complex genetic structure of Leopoldamys neilli is supported by congruent mitochondrial and nuclear markers and has been influenced by the geological history of Thailand during Plio-Pleistocene.

Revisiting the taxonomy of the Rattini tribe: a phylogeny-based delimitation of species boundaries.

BACKGROUND: Rodents are recognized as hosts for at least 60 zoonotic diseases and may represent a serious threat for human health. In the context of global environmental changes and increasing mobility of humans and animals, contacts between pathogens and potential animal hosts and vectors are modified, amplifying the risk of disease emergence. An accurate identification of each rodent at a specific level is needed in order to understand their implications in the transmission of diseases. Among the Muridae, the Rattini tribe encompasses 167 species inhabiting South East Asia, a hotspot of both biodiversity and emerging and re-emerging diseases. The region faces growing economical development that affects habitats, biodiversity and health. Rat species have been demonstrated as significant hosts of pathogens but are still difficult to recognize at a specific level using morphological criteria. DNA-barcoding methods appear as accurate tools for rat species identification but their use is hampered by the need of reliable identification of reference specimens. In this study, we explore and highlight the limits of the current taxonomy of the Rattini tribe. RESULTS: We used the DNA sequence information itself as the primary information source to establish group membership and estimate putative species boundaries. We sequenced two mitochondrial and one nuclear genes from 122 rat samples to perform phylogenetic reconstructions. The method of Pons and colleagues (2006) that determines, with no prior expectations, the locations of ancestral nodes defining putative species was then applied to our dataset. To give an appropriate name to each cluster recognized as a putative species, we reviewed information from the literature and obtained sequences from a museum holotype specimen following the ancient DNA criteria. CONCLUSIONS: Using a recently developed methodology, this study succeeds in refining the taxonomy of one of the most difficult groups of mammals. Most of the species expected within the area were retrieved but new putative species limits were also indicated, in particular within Berylmys and Rattus genera, where future taxonomic studies should be directed. Our study lays the foundations to better investigate rodent-born diseases in South East Asia and illustrates the relevance of evolutionary studies for health and medical sciences.

Ectoparasitic fauna of birds, and volant and non-volant small mammals captured at Srinakarin Dam, Kanchanaburi, Thailand.

The investigation of ectoparasitic fauna on birds, and volant and nonvolant small mammals at Srinakarin Dam, Kanchanaburi Province, Thailand was carried out under a national biodiversity and disease surveillance program for four consecutive months: January, February, May and June 2009. A total of 122 animals, comprised of 15 species of birds, 9 species of volant small mammals and 8 species of non-volant small mammals were examined for ectoparasite infestation. Of these animals, 1 genus of hard ticks (Ixodidae), 2 species of mesostigmatid mites (Laelapidae), 4 genera in three families of astigmatid mites (Proctophyllodidae, Pteronyssidae and Trouessartiidae), 4 species in three families of lice (Philopteridae, Polyplacidae and Trichodectidae) and 2 families of batflies (Nycteribiidae and Streblidae) were collected. This is the first survey conducted to determine ectoparasites infesting birds and small mammals living in the reserved forest of Srinakarin Dam, Thailand. A lower infestation rate of ectoparasites was observed in mammals, ranging from 3.5% to 10.3% than birds, with infestation rates between 7.3% and 34.2%. No major potential health risks to people who lived in this area were found.

Geographical variation in skull morphology of gray-bellied squirrel Callosciurus caniceps.

The geographical variation of the gray-bellied squirrel (Callosciurus caniceps) was examined using osteometry of skull in Southeast Asia. From the principal component analysis (PCA), the plots of the northern localities from Nan to Kanchanaburi and those of the southern localities from Narathiwat to Kuala Lumpur in male were completely separated. In female, the plots of the locality from Uttradit to Kanchanaburi and those of the locality from Pattani to Negri Sembilan were completely separated. We called these northern localities and southern localities which are distinguished by the PCA as N group and S group. The size and shape of the skulls of these squirrels indicated the differences between N group and S group from t-test and U-test. These results may be influenced by the two transitions of the phytogeography around the southernmost locality in N group and the northernmost locality in S group in the peninsular Thailand and Malay Peninsula. Localities which are located between N and S groups were called the Middle (M) group. From the PCA among N, S groups and each locality of M group, the plots of localities such as Prachuap Khiri Khan, Chumphon, Krabi, Nakhon Si Thammarat and Trang in both sexes of M group could not be separated from those of N and S groups. We suggest that the sympatric distribution of N and S groups and the hybrid of N and S populations may be seen in these localities of M group.