Using the 2020 global pandemic as a springboard to highlight the need for amphibian conservation in eastern Asia.

Emerging infectious diseases are on the rise in many different taxa, including, among others, the amphibian batrachochytrids, the snake fungal disease and the Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) virus, responsible for Coronavirus disease 2019 (COVID-19) in mammals. Following the onset of the pandemic linked to COVID-19, eastern Asia has shown strong leadership, taking actions to regulate the trade of potential vector species in several regions. These actions were taken in response to an increase in public awareness, and the need for a quick reaction to mitigate against further pandemics. However, trade restrictions rarely affect amphibians, despite the risk of pathogen transmission, directly, or indirectly through habitat destruction and the loss of vector consumption. Thus, species that help alleviate the risk of zoonoses or provide biological control are not protected. Hence, in view of the global amphibian decline and the risk of zoonoses, we support the current wildlife trade regulations and support measures to safeguard wildlife from overexploitation. The current period of regulation overhaul should be used as a springboard for amphibian conservation. To mitigate risks, we suggest the following stipulations specifically for amphibians. I) Restrictions to amphibian farming in eastern Asia, in relation to pathogen transmission and the establishment of invasive species. II) Regulation of the amphibian pet trade, with a focus on potential vector species. III) Expansion of the wildlife trade ban, to limit the wildlife-human-pet interface. The resulting actions will benefit both human and wildlife populations, as they will lead to a decrease in the risk of zoonoses and better protection of the environment. SIGNIFICANCE STATEMENT: There is an increasing number of emerging infectious diseases impacting all species, including amphibians, reptiles and mammals. The latest threat to humans is the virus responsible for COVID-19, and the resulting pandemic. Countries in eastern Asia have taken steps to regulate wildlife trade and prevent further zoonoses thereby decreasing the risk of pathogens arising from wild species. However, as amphibians are generally excluded from regulations we support specific trade restrictions: I) Restrictions to amphibian farming; II) regulation of the amphibian pet trade; III) expansion of the wildlife trade ban. These restrictions will benefit both human and wildlife populations by decreasing the risks of zoonoses and better protecting the environment.

Synonymous single nucleotide polymorphism in arsenic (+3) methyltransferase of the Western mosquitofish (Gambusia affinis) and its gene expression among field populations.

Naturally occurring arsenic is toxic at extremely low concentrations, yet some species persist even in high arsenic environments. We wanted to test if these species show evidence of evolution associated with arsenic exposure. To do this, we compared allelic variation across 872 coding nucleotides of arsenic (+3) methyltransferase (as3mt) and whole fish as3mt gene expression from three field populations of Gambusia affinis, from water sources containing low (1.9 ppb), medium-low (3.3 ppb), and high (15.7 ppb) levels of arsenic. The high arsenic site exceeds the US EPA's Maximum Contamination Level for drinking water. Medium-low and high populations exhibited homozygosity, and no sequence variation across all animals sampled. Eleven of 24 fish examined (45.8%) in the low arsenic population harbored synonymous single nucleotide polymorphisms (SNPs) in exons 4 and/or 10. SNP presence in the low arsenic population was not associated with differences in as3mt transcript levels compared to fish from the medium-low site, where SNPs were noted; however, as3mt expression in fish from the high arsenic concentration site was significantly lower than the other two sites. Low sequence variation in fish populations from sites with medium-low and high arsenic concentrations suggests greater selective pressure on this allele, while higher variation in the low population suggests a relaxed selection. Our results suggest gene regulation associated with arsenic detoxification may play a more crucial role in influencing responses to arsenic than polymorphic gene sequence. Understanding microevolutionary processes to various contaminants require the evaluation of multiple populations across a wide range of pollution exposures.

Interactions between a small chronic increase in diel water temperature and exposure to a common environmental contaminant on development of Arizona tiger salamander larvae.

Global climate change leading to increased temperatures may affect shifts in physiological processes especially in ectothermic organisms. Temperature-dependent shifts in developmental rate in particular, may lead to life-long changes in adult morphology and physiology. Combined with anthropogenic changes in the chemical environment, changes in developmental outcomes may affect adult functionality. The purpose of this study is to determine 1) if small increases in diel water temperature affect the development of Arizona tiger salamander (Ambystoma tigrinum nebulosum) larvae, and 2) if this change interacts with exposure to the common environmental thyroid disrupting compound, perchlorate. Larvae between Watson and Russell developmental stages 8-13 were exposed to ammonium perchlorate (AP) at doses of 0, 20 or 200ppb and then raised at either ambient or a 0.9°C elevated above ambient temperature for 81days in outdoor enclosures. During the first 5 treatment weeks, AP treatment induced slower development and smaller snout-vent length (SVL) of exposed larvae, but only in the elevated temperature group. During the later stages of development, the small increase in temperature, regardless of AP treatment, tended to decrease the time to metamorphosis and resulted in a significantly smaller body mass and worse body condition. Our results suggest that even small diel water temperature increases can affect the developmental process of salamanders and this shift in the water temperature may interact with a common environmental contaminant.

Population genetic structure of endangered Mongolian racerunner (Eremias argus) from the Korean Peninsula.

The Mongolian racerunner (Eremias argus) is a small lacertid lizard species, and its distribution range encompasses the Korean Peninsula, Mongolia, China and Russia. Eremias argus is widespread, but populations on the Korean Peninsula are small and declining, provoking concerns that genetic diversity is being lost. This species is currently listed under the Protection of Wild Fauna and Flora Act in South Korea. In this study, nine novel microsatellites for E. argus were developed with a biotin-enrichment method and used to understand its population genetic structure and delineate conservation units on the Korean Peninsula. Overall, low intrapopulation genetic diversity was observed (mean number of alleles per locus = 2.463; mean H E = 0.398) from 10 populations investigated (n = 110). Two populations (among five with n≥ 10) showed an excess of heterozygosity expected under HWE relative to that expected at mutation-drift equilibrium, indicating severe reduction in population sizes. With only a few exceptions, the overall genetic differentiation among populations was substantial with the high levels of pairwise-F ST (0.006-0.746) and -R ST (0.034-0.940) values. The results of Bayesian STRUCTURE analysis showed that E. argus populations on the Korean Peninsula were most likely partitioned into three genetic clusters. Taken all together, such low levels of gene flow and strong genetic structuring have critical implications for the conservation of this endangered species and its management.

First Report of Endemic Frog Virus 3 (FV3)-like Ranaviruses in the Korean Clawed Salamander (Onychodactylus koreanus) in Asia.

Frog virus 3 (FV3) in the genus Ranavirus of the family Iridoviridae causes mass mortality in both anurans and urodeles worldwide; however, the phylogenetic origin of FV3-like ranaviruses is not well established. In Asia, three FV3-like ranaviruses have been reported in farmed populations of amphibians and reptiles. Here, we report the first case of endemic FV3-like ranavirus infections in the Korean clawed salamander Onychodactylus koreanus, caught in wild mountain streams in the Republic of Korea (ROK), through whole-genome sequencing and phylogenetic analysis. Two isolated FV3-like ranaviruses (Onychodactylus koreanus ranavirus, OKRV1 and 2) showed high similarity with the Rana grylio virus (RGV, 91.5%) and Rana nigromaculata ranavirus (RNRV, 92.2%) but relatively low similarity with the soft-shelled turtle iridovirus (STIV, 84.2%) in open reading frame (ORF) comparisons. OKRV1 and 2 formed a monophyletic clade with previously known Asian FV3-like ranaviruses, a sister group of the New World FV3-like ranavirus clade. Our results suggest that OKRV1 and 2 are FV3-like ranaviruses endemic to the ROK, and RGV and RNRV might also be endemic strains in China, unlike previous speculation. Our data have great implications for the study of the phylogeny and spreading routes of FV3-like ranaviruses and suggest the need for additional detection and analysis of FV3-like ranaviruses in wild populations in Asian countries.

Identifying the source rookery of green turtles (Chelonia mydas) found in feeding grounds around the Korean Peninsula.

Determining the genetic diversity and source rookeries of sea turtles collected from feeding grounds can facilitate effective conservation initiatives. To ascertain the genetic composition and source rookery, we examined a partial sequence of the mitochondrial control region (CR, 796 bp) of 40 green turtles (Chelonia mydas) collected from feeding grounds around the Korean Peninsula between 2014 and 2022. We conducted genetic and mixed-stock analyses (MSA) and identified 10 CR haplotypes previously reported in Japanese populations. In the haplotype network, six, three, and one haplotype(s) grouped with the Japan, Indo-Pacific, and Central South Pacific clades, respectively. The primary rookeries of the green turtles were two distantly remote sites, Ogasawara (OGA) and Central Ryukyu Island (CRI), approximately 1,300 km apart from each other. Comparing three parameters (season, maturity, and specific feeding ground), we noted that OGA was mainly associated with summer and the Jeju Sea, whereas CRI was with fall and the East (Japan) Sea ground. The maturity did not show a distinct pattern. Our results indicate that green turtles in the feeding grounds around the Korean Peninsula originate mainly from the Japan MU and have genetic origins in the Japan, Indo-Pacific, and Central South Pacific clades. Our results provide crucial insights into rookeries and MUs, which are the focus of conservation efforts of the Republic of Korea and potential parties to collaborate for green turtle conservation.

Habitat use of loggerhead (Caretta caretta) and green (Chelonia mydas) turtles at the northern limit of their distribution range of the Northwest Pacific Ocean.

Verifying habitats, including the foraging and nesting areas for sea turtles, enables an understanding of their spatial ecology and successful planning of their conservation and management strategies. Recently, the observation frequency and bycatch of loggerhead (Caretta caretta) and green (Chelonia mydas) turtles have increased in the northern limit of their distribution range, in the northern part of the East China Sea and East (Japan) Sea. We conducted satellite tracking to investigate the habitat use of seven loggerhead and eight green turtles from June 2016 to August 2022 in this area, where little is known about their spatial ecology. We applied a 50 percent volume contour method to determine their main foraging areas and analyzed 6 environmental variables to characterize their habitats. Loggerhead turtles mainly stayed in and used the East China Sea as a foraging area during the tracking period, while two individuals among them also used the East Sea as a seasonal foraging area. Most green turtles also used the East China Sea as a foraging area, near South Korea and Japan, with one individual among them using the lower area of the East Sea as a seasonal foraging area. Notably, one green turtle traveled to Hainan Island in the South China Sea, a historical nesting area. Our results showed that the two sea turtle species included the East Sea as a seasonal foraging area, possibly owing to the abundance of food sources available, despite its relatively lower sea temperature. Considering that loggerhead and green sea turtles were observed using the northern part of the East China Sea and East Sea more frequently than previously known and that the sea temperature gradually increases due to climate change, conservation and management activities are required for sea turtles in these areas.

Arginine vasotocin (AVT) triggers courtship behavior without exposure to external stimuli and modulates the olfactory response of male Hynobius leechii salamanders.

To study the function of arginine vasotocin (AVT) in a specific courtship behavior (body undulation) in male Hynobius leechii, we injected various doses of AVT or an AVT V1a or V2 receptor antagonist into breeding and non-breeding males. After these injections, we placed the males alone or with breeding females in Petri dishes and measured the incidence and frequency of body undulation. Additionally, to test whether AVT modulates the olfactory response of males, we exposed breeding males that were injected with AVT to female odors and measured the same response. Both breeding and non-breeding males intraperitoneally injected with 50 or 100 µg of AVT exhibited body undulation. Additionally, breeding males intraperitoneally injected with 50 or 100 µg of AVT exhibited an increased frequency of body undulation when exposed to female odors. The intraperitoneal injection of 5, 25, or 50 µg of the AVT V1a or V2 receptor antagonist did not significantly decrease the incidence or frequency of body undulation of 100 µg AVT-injected breeding males. However, in a post-hoc experiment, the breeding males that were subcutaneously injected with 100 µg of the AVT V1a receptor antagonist exhibited a significant decrease in the frequency of body undulation, induced by exposure to females. Additionally, a central injection of 1 µg of AVT into the brain induced body undulation in breeding males. Our results show that AVT not only induces a specific courtship behavior in male H. leechii via AVT V1a receptors but also modulates the olfactory response.

Daily temperature fluctuations unpredictably influence developmental rate and morphology at a critical early larval stage in a frog.

BACKGROUND: Environmental temperature has profound consequences for early amphibian development and many field and laboratory studies have examined this. Most laboratory studies that have characterized the influence of temperature on development in amphibians have failed to incorporate the realities of diel temperature fluctuations (DTF), which can be considerable for pond-breeding amphibians. RESULTS: We evaluated the effects of different ecologically relevant ranges of DTF compared with effects of constant temperatures on development of embryos and larvae of the Korean fire-bellied toad (Bombina orientalis). We constructed thermal reaction norms for developmental stage, snout-vent length, and tail length by fitting a Gompertz-Gaussian function to measurements taken from embryos after 66 hours of development in 12 different constant temperature environments between 14°C and 36°C. We used these reaction norms as null models to test the hypothesis that developmental effects of DTF are more than the sum of average constant temperature effects over the distribution of temperatures experienced. We predicted from these models that growth and differentiation would be positively correlated with average temperature at low levels of DTF but not at higher levels of DTF. We tested our prediction in the laboratory by rearing B. orientalis embryos at three average temperatures (20°C, 24°C, and 28°C) and four levels of thermal variation (0°C, 6°C, 13°C, and 20°C). Several of the observed responses to DTF were significantly different from both predictions of the model and from responses in constant temperature treatments at the same average temperatures. At an average temperature of 24°C, only the highest level of DTF affected differentiation and growth rates, but at both cooler and warmer average temperatures, moderate DTF was enough to slow developmental and tail growth rates. CONCLUSIONS: These results demonstrate that both the magnitude of DTF range and thermal averages need to be considered simultaneously when parsing the effects of changing thermal environments on complex developmental responses, particularly when they have potential functional and adaptive significance.

Distribution and habitat assessments of the Slender racer, Orientocoluber spinalis, for the registration of nationally endangered species in the Republic of Korea.

Conservation assessments are essential for preserving biodiversity. However, many reptile species have not been evaluated owing to data deficiencies. The Slender racer (Orientocoluber spinalis) is threatened in four out of six inhabiting countries. However, despite its apparent rarity and data deficiency, the International Union for Conservation of Nature (IUCN) has classified it as a Least Concern. In this study, we combined field surveys, habitat analysis, and ecological niche models (ENMs) to identify the critical habitat characteristics of O. spinalis, evaluate its distribution status in the Republic of Korea, and register it as a nationally endangered species. Across the country, we found a few small populations on the mainland but large populations on the islands. Orientocoluber spinalis is mainly found in low-altitude ecotone habitats between grasslands and forests. Based on previous genetic and climatic studies, we propose designating it as an endangered species to conserve this species in protected areas such as national parks, and its non-isolated mainland populations can be preserved as source populations.

Large-scale habitat association modeling of the endangered Korean ratsnake (Elaphe schrenckii).

We used radio-telemetric monitoring and statistical models to investigate habitat associations of the endangered Korean ratsnake (Elaphe schrenckii) in a mountain region of Woraksan National Park based on the data collected from July 2007 to April 2009. We evaluated a priori models at landscape scales that incorporated natural and artificial variables to explain ratsnake presence and absence using geographic information system (GIS) databases. We generated models using logistic regression combined with Akaike's information criterion (AIC) to determine which variables are most important. The best-ranked model predicted that the ratsnakes are likely to use areas with high annual solar radiation and that they are closer to streams, coniferous and mixed forests, farmland, houses, and barren ground. We next evaluated the model by applying it to an internal and an external dataset, where the model showed high power of prediction and discrimination on the internal dataset with estimated mean probability of 94% while showing suitability as a predictive model on the external dataset with 64% probability. The present study suggests that a created habitat model can provide valuable contributions to conservation and management efforts directed towards ratsnake populations.

Prevalence of Ranavirus Infection in Three Anuran Species across South Korea.

To cope with amphibian die-offs caused by ranavirus, it is important to know the underlying ranavirus prevalence in a region. We studied the ranavirus prevalence in tadpoles of two native and one introduced anuran species inhabiting agricultural and surrounding areas at 49 locations across eight provinces of South Korea by applying qPCR. The local ranavirus prevalence and the individual infection rates at infected locations were 32.6% and 16.1%, respectively, for Dryophytes japonicus (Japanese tree frog); 25.6% and 26.1% for Pelophylax nigromaculatus (Black-spotted pond frog); and 30.5% and 50.0% for Lithobates catesbeianus (American bullfrog). The individual infection rate of L. catesbeianus was significantly greater than that of D. japonicus. The individual infection rate of P. nigromaculatus was related to the site-specific precipitation and air temperature. The individual infection rate gradually increased from Gosner development stage 39, and intermittent infection was confirmed in the early and middle developmental stages. Our results show that ranavirus is widespread among wild amphibians living in agricultural areas of South Korea, and mass die-offs by ranavirus could occur at any time.

A comparison of gene organisations and phylogenetic relationships of all 22 squamate species listed in South Korea using complete mitochondrial DNA.

Studies using complete mitochondrial genome data have the potential to increase our understanding on gene organisations and evolutionary species relationships. In this study, we compared complete mitochondrial genomes between all 22 squamate species listed in South Korea. In addition, we constructed Maximum Parsimony (MP), Maximum Likelihood (ML) and Bayesian Inference (BI) phylogenetic trees using 13 mitochondrial protein-coding genes. The mitochondrial genes for all six species in the suborder Sauria followed the same organisation as the sequenced Testudines (turtle) outgroup. In contrast, 16 snake species in the suborder Serpentes contained some gene organisational variations. For example, all snake species contained a second control region (CR2), while three species in the family Viperidae had a translocated tRNA-Pro gene region. In addition, the snake species, Elapheschrenckii, carried a tRNA-Pro pseudogene. We were also able to identify a translocation of a tRNA-Asn gene within the five tRNA (WANCY gene region) gene clusters for two true sea snake species in the subfamily Hydrophiinae. Our BI phylogenetic tree was also well fitted against currently known Korean squamate phylogenetic trees, where each family and genus unit forms monophyletic clades and the suborder Sauria is paraphyletic to the suborder Serpentes. Our results may form the basis for future northeast Asian squamate phylogenetic studies.

Past, present, and future predictions on the suitable habitat of the Slender racer (Orientocoluber spinalis) using species distribution models.

Species distribution models (SDMs) across past, present, and future timelines provide insights into the current distribution of these species and their reaction to climate change. Specifically, if a species is threatened or not well-known, the information may be critical to understand that species. In this study, we computed SDMs for Orientocoluber spinalis, a monotypic snake genus found in central and northeast Asia, across the past (last interglacial, last glacial maximum, and mid-Holocene), present, and future (2070s). The goal of the study was to understand the shifts in distribution across time, and the climatic factors primarily affecting the distribution of the species. We found the suitable habitat of O. spinalis to be persistently located in cold-dry winter and hot summer climatic areas where annual mean temperature, isothermality, and annual mean precipitation were important for suitable habitat conditions. Since the last glacial maximum, the suitable habitat of the species has consistently shifted northward. Despite the increase in suitable habitat, the rapid alterations in weather regimes because of climate change in the near future are likely to greatly threaten the southern populations of O. spinalis, especially in South Korea and China. To cope with such potential future threats, understanding the ecological requirements of the species and developing conservation plans are urgently needed.

Movement Patterns of Juvenile Loggerhead Turtles (Caretta caretta L. 1758) and Green Turtles (Chelonia mydas L. 1758) Hatched in Captivity and Released in the Korean Waters.

With most sea turtle populations declining, activities to conserve their habitat and nesting grounds and restore their populations are being implemented worldwide. To preserve the Northwestern Pacific populations, the National Marine Biodiversity Institute of Korea has been releasing artificially propagated sea turtles, but whether these individuals join the wild population remains unknown. The present study aimed to determine the movement patterns of artificially propagated juvenile loggerhead (Caretta caretta) and green (Chelonia mydas) turtles fitted with satellite transmitters on their carapaces and released in the waters of Jeju or Yeosu, Republic of Korea, between August 2018 and April 2022. Loggerheads traveled northward to the East Sea, whereas green turtles moved west or southwest. Two 36-month-old and two 48-month-old loggerheads moved toward their potential nursery grounds and toward their feeding grounds, respectively. Three green turtles with a curved carapace length (CCL) of <40 cm moved toward their nursery or feeding grounds, while three individuals (CCL > 45 cm) moved toward their inshore foraging areas. The travel paths were closely related to the direction of local sea currents. Our results implied that releasing artificially propagated sea turtles, considering their age and CCL, can positively contribute to the conservation of Northwestern Pacific populations.

Habitat use and movement patterns of the viviparous aquatic snake, Oocatochus rufodorsatus, from northeast Asia.

To determine the habitat usage and movement patterns of the viviparous aquatic snake Oocatochus rufodorsatus (formerly Elaphe rufodorsata), we radio-tracked 21 snakes on agricultural lands during two active seasons in 2007 and 2008. Male and female snakes stayed close to aquatic habitats such as paddy fields and agricultural ponds during both breeding and non-breeding periods, except when the snakes moved to dry terrestrial areas to hibernate in late fall. The use of different structural features in the habitat, such as ground, tree, underground, and water, varied depending on the air and water temperatures, female's reproductive conditions, and the time of day. Male and female snakes moved about 17 m daily and postpartum females moved farther than antepartum females. The home ranges of males and females were 0.45 ha and 0.47 ha, respectively, and the year-round home range of this species was approximately 1.54 ha (95% fixed kernel). Thus, to conserve a population of O. rufodorsatus in our study area, areas including both aquatic and terrestrial habitats within a radius of 150 m from a core pond habitat must be preserved.

Habitat associations of the endangered gold-spotted pond frog (Rana chosenica).

We used field surveys and statistical models to investigate habitat associations of the endangered gold-spotted pond frog (Rana chosenica). The characteristics of its habitat are of great importance for effective conservation for this declining species in western South Korea. We evaluated a priori models that incorporated biotic and abiotic variables at the pond and landscape scales. The best-ranked model predicts that gold-spotted pond frogs will be more abundant at sites with fewer introduced American bullfrogs (Rana [= Lithobates] catesbeianus) and greater coverage of shallow, vegetated water. Our study leads us to conclude that limiting the spread and abundance of bullfrogs has the potential to aid conservation of the gold-spotted pond frog in our region.

Genetic diversity and inferred dispersal history of the Schlegel's Japanese Gecko (Gekko japonicus) in Northeast Asia based on population genetic analyses and paleo-species distribution modelling.

To understand the genetic diversity and dispersal history of Schlegel's Japanese gecko (Gekko japonicus), we performed genetic analyses and paleo-species distributional modelling. For the genetic analysis, we analyzed mitochondrial DNA (mtDNA) (cytochrome b [Cytb] and NADH dehydrogenase 2 [ND2]) and seven microsatellite loci of 353 individuals from 11 populations (2 east coast China, 4 west and central coast Japan and 5 Korea). For the paleo-species distribution modelling, we used 432 occurrence data points (125 China, 291 Japan and 16 Korea) over the Pleistocene and Holocene. China is inferred to be the source population, which had higher genetic diversity (mtDNA) and more private alleles (mtDNA) compared to Japanese and Korean populations. Differences between the three counties were very small in the mtDNA haplotype network despite some genetic structure among the three countries. Microsatellite analysis inferred that genetic exchange has actively occurred among the Chinese, Japanese and Korean populations. Suitable habitats in Japan should have been plentiful by the mid-Holocene, but have only recently become available in Korea. These results suggest that dispersal of G. japonicus occurred after the Holocene warming from the east coast of China to the west and central coasts of Japan and Korea, and gene flow is actively occurring among the three countries.

F-series prostaglandin function as sex pheromones in the Korean salamander, Hynobius leechii.

In order to test whether prostaglandins (PGs) function as sex pheromones in Hynobius leechii, a salamander that externally fertilizes its eggs, we conducted electro-olfactogram (EOG) studies with 19 PGs, liquid chromatography tandem mass spectrometry (LC-MS/MS) analyses of female and male holding waters, and behavioral tests on selected PGs. Of the 19 PGs tested, only three induced strong EOG responses from both males and ovulated females: 15-epi-prostaglandin F2alpha (15(R)-PGF2alpha), 15-keto-prostaglandin F2alpha (15K-PGF2alpha), and 13,14-dihydro-15-keto-prostaglandin F2alpha (13,14-dh-15K-PGF2alpha). In the LC-MS/MS studies, samples of holding water from ovulated females contained higher concentrations of 15(R)-PGF2alpha, PGF2alpha, and 13,14-dh-15K-PGF2alpha than those from males or oviposited females. In the behavioral tests, only 15(R)-PGF2alpha and ovulated female holding water induced significant reproductive behavior from male salamanders. These results suggest that F-series prostaglandins function as sex pheromones in amphibians.

Spring and summer microhabitat use by Schlegel's Japanese gecko, Gekko japonicus (Reptilia: Squamata: Gekkonidae), in urban areas.

Differential microhabitat use may be beneficial to achieving fitness in seasonally variable environmental conditions. To explore whether the microhabitat use of the nocturnal Schlegel's Japanese gecko, Gekko japonicus, varies seasonally and depends on juvenile, male, and female reproductive groups, we investigated five categorical and five quantitative measure variables of microhabitat use in a wild population both in spring and summer. Most geckos were found on white, vertical planes of concrete and plastered brick walls. None of the categorical variables (type of location, substrate, substrate color, light source, and refuge) significantly differed according to season or group, while substrate temperature and irradiance at the location where geckos were observed and the distance from the nearest potential refuge were significantly greater in summer than in spring. The quantitative measure variables did not differ among the reproductive groups. These results suggest that G. japonicus seasonally adjusts its microhabitat use mainly in terms of quantitative measure variables rather than categorical variables.