Blood clinical biochemistry and packed cell volume of the Chinese three-striped box turtle, Cuora trifasciata (Reptilia: Geoemydidae).

Reference intervals for blood biochemical analytes and packed cell volume are useful to veterinarians and conservation programs for monitoring the health of threatened species. Populations of the critically endangered Chinese three-striped box turtle, Cuora trifasciata (Bell, 1825), are heavily depleted in the wild because of overharvesting; and many individuals are now maintained in captive breeding programs. Reference intervals of biochemical analytes and packed cell volume were established for individuals of this species held in a captive breeding program at Kadoorie Farm and Botanic Garden, Hong Kong. Blood samples were obtained from 86 healthy individuals from a captive population during annual health examinations between 2006 and 2010. Packed cell volume and 17 biochemical analytes were measured, and data were analyzed using both multivariate and univariate statistical analyses. Significant differences in eight analytes between males and females were identified. Ten analytes were found to increase or decrease significantly with the body weight of the turtles. The reported plasma reference intervals, based on 2.5th and 97.5th percentiles, may serve as benchmarks for clinical assessment and conservation of this critically endangered species.

Demographic evidence of illegal harvesting of an endangered asian turtle.

Harvesting pressure on Asian freshwater turtles is severe, and dramatic population declines of these turtles are being driven by unsustainable collection for food markets, pet trade, and traditional Chinese medicine. Populations of big-headed turtle (Platysternon megacephalum) have declined substantially across its distribution, particularly in China, because of overcollection. To understand the effects of chronic harvesting pressure on big-headed turtle populations, we examined the effects of illegal harvesting on the demography of populations in Hong Kong, where some populations still exist. We used mark-recapture methods to compare demographic characteristics between sites with harvesting histories and one site in a fully protected area. Sites with a history of illegal turtle harvesting were characterized by the absence of large adults and skewed ratios of juveniles to adults, which may have negative implications for the long-term viability of populations. These sites also had lower densities of adults and smaller adult body sizes than the protected site. Given that populations throughout most of the species' range are heavily harvested and individuals are increasingly difficult to find in mainland China, the illegal collection of turtles from populations in Hong Kong may increase over time. Long-term monitoring of populations is essential to track effects of illegal collection, and increased patrolling is needed to help control illegal harvesting of populations, particularly in national parks. Because few, if any, other completely protected populations remain in the region, our data on an unharvested population of big-headed turtles serve as an important reference for assessing the negative consequences of harvesting on populations of stream turtles. Evidencia Demográfica de la Captura Ilegal de una Tortuga Asiática en Peligro.

The identity of Tokays (Squamata: Gekkonidae: Gekko) from Hong Kong.

The Tokay gecko (Gekko gecko), found throughout Southeast Asia and India, is a heavily harvested species of high commercial value. Recent studies have supported the elevation of the black morph of Tokay gecko, found only in southern China and northern Vietnam, to the species Gekko reevesii. Previous genetic studies focused on specimens of G. reevesii from southwestern populations. Hong Kong, in southeastern China, has native populations of G. reevesii. To verify the identity of G. reevesii in Hong Kong, we employed three mitochondrial genes (COI, Cytb & ND2) and constructed a matrilineal genealogy using other specimens from Guangxi (southwestern China) and northern Vietnam, as well as G. gecko from a wide range of Southeast Asian countries. Our study confirmed that G. reevesii occurs naturally in Hong Kong, but one exotic population of G. gecko, likely a translocation from international trade, was also revealed. Our study did not reject the species ranking of G. reevesii. Moreover, like previous studies, we recovered a paraphyletic G. gecko, which may reflect a species complex, hybridization or incomplete lineage sorting. More extensive sampling of the two species over a broader range of their asserted distribution together with the use of both mitochondrial and nuclear DNA are required to better investigate their biogeography.

Developing bottom drifters to better understand the stranding locations of cold-stunned sea turtles in Cape Cod Bay, Massachusetts.

Every fall, juvenile sea turtles in the Northwest Atlantic Ocean are threatened by rapidly declining water temperatures. When sea turtles become hypothermic, or cold-stunned, they lose mobility-either at the surface, subsurface, or the bottom of the water column-and eventually strand at the shoreline where rescue teams associated with the Sea Turtle Stranding and Salvage Network may search for them. Understanding the effects of ocean currents on the potential stranding locations of cold-stunned sea turtles is essential to better understand stranding hotspots and increase the probability of successful discovery and recovery of turtles before they die in the cold temperatures. Traditional oceanographic drifters-instruments used to track currents-have been used to examine relationships between current and stranding locations in Cape Cod Bay, but these drifters are not representative of sea turtle morphology and do not assess how bottom currents affect stranding locations. To address these knowledge gaps, we designed new drifters that represent the shape and dimensions of sea turtles-one that can float at the surface and one that sinks to the bottom-to track both surface and bottom currents in Cape Cod Bay. We found a marked difference between the trajectories of our new drifter models and those that were previously used for similar research. These findings bring us one step closer to identifying the transport pathways for cold-stunned sea turtles and optimizing cold-stunned sea turtle search and rescue efforts in Cape Cod.

Online sale of small turtles circumvents public health regulations in the United States.

In the United States (U.S.), pet turtles have been associated with outbreaks of salmonellosis, a serious and sometimes-fatal intestinal illness caused by Salmonella bacteria, with nearly 300,000 people being infected in some years. Children are particularly susceptible because of their propensity to put items, including small turtles, in their mouths. In 1975, a U.S. federal regulation prohibited the sale of turtles <4 inches (101.6 mm) in size, except for the purposes of export, scientific, or educational purposes. This regulation was established to reduce the incidence of salmonellosis, particularly in small children. Previous research has not evaluated the availability of turtles <4 inches in size on websites selling wildlife. We monitored 16 websites in 2021 and quantified listings of small turtles. We determined whether information on Salmonella, the 1975 federal regulation, or related state regulations were provided on the websites and determined legality of sales of small turtles by state regulations. We found that all 16 websites openly advertised and sold turtles <4 inches in size, but only half of these websites provided information about Salmonella and/or the federal regulation. These websites required buyers to confirm that they were not purchasing a turtle as a pet, thereby putting the onus on the consumer to adhere to the regulation. We documented 515 listings of turtles <4 inches in size, including 47 species and one hybrid. Our study has demonstrated that internet sales of small turtles currently represent part of the thriving online pet trade in the U.S. Enforcement of the federal regulation faces jurisdictional challenges in most states. Therefore, we recommend continued public education campaigns by public health agencies in the U.S. to help reduce the risk that pet turtle ownership presents.

Non-marine turtle plays important functional roles in Indonesian ecosystems.

The Southeast Asian box turtle (Cuora amboinensis) is numerically the most important turtle exported from Indonesia. Listed as Vulnerable by the IUCN, this turtle is heavily harvested and exported for food and traditional medicine in China and for the pet trade primarily in the United States, Europe, and Japan. Despite its significance in global markets, relatively little is known about the species' ecology or importance to ecosystems. We conducted our research in a national park in Sulawesi, Indonesia, and our objectives were to quantify trophic breadth, capacity for seed dispersal between aquatic and terrestrial ecosystems, and whether ingestion of seeds by C. amboinensis enhances germination. We obtained diet samples from 200 individual turtles and found that the species is omnivorous, exhibiting an ontogenetic shift from more carnivorous to more omnivorous. Both subadults and adults scavenged on other vertebrates. In a seed passage experiment, turtles passed seeds for 2‒9 days after ingestion. Radio-tracked turtles moved, on average, about 35 m per day, indicating that seeds from ingested fruits, given seed passage durations, could be dispersed 70‒313 m from the parent tree and potentially between wetland and upland ecosystems. In a seed germination experiment, we found that ingestion of seeds by turtles enhanced germination, as compared with control seeds, for four of six plant species tested. Of these, two are common in the national park, making up a significant proportion of plant biomass in lowland swamp forest and around ephemeral pools in savanna, and are highly valued outside of the park for their lumber for construction of houses, furniture, and boats. Protection of C. amboinensis populations may be important for maintaining trophic linkages that benefit biodiversity, communities, and local economies.

Effect of road deicing salt on the susceptibility of amphibian embryos to infection by water molds.

Some causative agents of amphibian declines act synergistically to impact individual amphibians and their populations. In particular, pathogenic water molds (aquatic oomycetes) interact with environmental stressors and increase mortality in amphibian embryos. We documented colonization of eggs of three amphibian species, the wood frog (Rana sylvatica), the green frog (Rana clamitans), and the spotted salamander (Ambystoma maculatum), by water molds in the field and examined the interactive effects of road deicing salt and water molds, two known sources of mortality for amphibian embryos, on two species, R. clamitans and A. maculatum in the laboratory. We found that exposure to water molds did not affect embryonic survivorship in either A. maculatum or R. clamitans, regardless of the concentration of road salt to which their eggs were exposed. Road salt decreased survivorship of A. maculatum, but not R. clamitans, and frequency of malformations increased significantly in both species at the highest salinity concentration. The lack of an effect of water molds on survival of embryos and no interaction between road salt and water molds indicates that observations of colonization of these eggs by water molds in the field probably represent a secondary invasion of unfertilized eggs or of embryos that had died of other causes. Given increasing salinization of freshwater habitats on several continents and the global distribution of water molds, our results suggest that some amphibian species may not be susceptible to the combined effects of these factors, permitting amphibian decline researchers to devote their attention to other potential causes.

Population demography of Oldham's leaf turtle (Cyclemys oldhamii) in protected and disturbed habitats in Thailand.

BACKGROUND: Freshwater turtle populations are vulnerable to a range of human activities because of particular life history attributes, and anthropogenic impacts can cause shifts in demographic traits, including survival, density and population structure. Asian freshwater turtles have undergone dramatic population declines in recent decades principally because of collection for food, pet, and traditional medicine markets. Despite this, few studies have been conducted on the population demography of these turtles, thereby limiting our understanding of population trends and the development of conservation actions. Oldham's leaf turtle (Cyclemys oldhamii) is one of the most commonly traded turtles in Asian markets, but previous published studies have focused solely on systematics. METHODS: We conducted a mark-recapture study of C. oldhamii at three sites in northeastern Thailand-a protected stream, a degraded stream, and human-constructed ponds-and evaluated differences in survival, density, population structure, and sexual dimorphism among sites. RESULTS: We captured 77 turtles at the protected stream, 67 at the constructed ponds, and two in the degraded stream. Survival was 12% lower and density was 35% lower in the constructed ponds than in the protected stream. Size class structure was skewed toward smaller individuals at the constructed ponds, and both sites exhibited subadult-skewed age class structure. Sex ratios were not statistically different than 1:1 at either site and did not differ between sites. We did not document sexual dimorphism in either population. DISCUSSION: Explanations for lower survival, lower densities, and skewed size class structure at the constructed ponds include collection for consumption or Buddhist prayer release locally, collection for illegal export from Thailand, predation by domestic dogs associated with humans living nearby, or lower habitat quality. Evidence from our study suggests that collection, either for local use or export, is the most likely explanation for differences in demographic characteristics between the two sites. The information gained from this study may contribute to a status assessment for C. oldhamii and development of conservation actions should they become necessary to protect populations in Thailand.

Impacts of road deicing salt on the demography of vernal pool-breeding amphibians.

Deicing agents, primarily road salt, are applied to roads in 26 states in the United States and in a number of European countries, yet the scale of impacts of road salt on aquatic organisms remains largely under-studied. The issue is germane to amphibian conservation because both adult and larval amphibians are known to be particularly sensitive to changes in their osmolar environments. In this study, we combined survey, experimental, and demographic modeling approaches to evaluate the possible effects of road salt on two common vernal-pond-breeding amphibian species, the spotted salamander (Ambystoma maculatum) and the wood frog (Rana sylvatica). We found that in the Adirondack Mountain Region of New York (USA), road salt traveled up to 172 m from the highway into wetlands. Surveys showed that egg mass densities of spotted salamanders (A. maculatum) and wood frogs (R. sylvatica) were two times higher in forest pools than roadside pools, but this pattern was better explained by road proximity than by increased salinity. Experiments demonstrated that embryonic and larval survival were reduced at moderate (500 muS) and high conductivities (3000 muS) in A. maculatum and at high conductivities in R. sylvatica. Demographic models suggest that such egg and larval stage effects of salt may have important impacts on populations near roads, particularly in the case of A. maculatum, for which salt exposure may lead to local extinction. For both species, the effect of road salt was dependent upon the strength of larval density dependence and declined rapidly with distance from the roadside, with the greatest negative effects being limited to within 50 m. Based on this evidence, we argue that efforts to protect local populations of A. maculatum and R. sylvatica in roadside wetlands should, in part, be aimed at reducing application of road salt near wetlands with high conductivity levels.

Signals of forest degradation in the demography of common Asian amphibians.

BACKGROUND: Lowland areas in tropical East and Southeast Asia have a long history of conversion from forestland to agricultural land, with many remaining forests being chronically degraded by wood cutting, livestock grazing, and burning. Wetland-breeding amphibians that have evolved in lowland forests in the region have adjusted to changes in habitat composition caused by humans' activities, and populations continue to persist. However, we have little understanding of the impacts of forest disturbance on these species beyond assessments of abundance and distribution, and species considered to be common and widespread have been largely neglected. METHODS: We examined body condition and sex ratios of toads (Duttaphrynus melanostictus), predation risk in treefrogs (2 Polypedates spp.), and growth and survival of leaf litter frogs (2 Microhyla spp.) in agricultural land, degraded forest, and intact forest in two study areas, Thailand and Hong Kong. RESULTS: Toad populations exhibited higher body condition and female-biased sex ratios in intact forest. Predation of treefrog embryos by flies was lower in intact and degraded forests than in agricultural land. Embryonic survival and larval growth and survival in leaf litter frogs were lower in intact forests than in agricultural land. Results for each study were similar between study areas. DISCUSSION: For three of five of these common amphibian species, we documented signals of forest loss and disturbance in their populations. Although these species occur in disturbed habitats, loss of forest cover continues to degrade aspects of their population demography. We urge conservation biologists to consider that populations of species appearing to be common, widespread, and tolerant of human disturbance may be eroding over time.

Does forest extent affect salamander survival? Evidence from a long-term demographic study of a tropical newt.

Forest loss has been associated with reduced survival in many vertebrates, and previous research on amphibians has mostly focused on effects at early life stages. Paramesotriton hongkongensis is a tropical newt that breeds in streams but spends up to 10 months per year in terrestrial habitats. Populations are threatened by habitat degradation and collection for the pet trade, but the cryptic terrestrial lifestyle of this newt has limited our understanding of its population ecology, which inhibits development of a species-specific conservation plan. We conducted an eight-year (2007-2014) mark-recapture study on four P. hongkongensis populations in Hong Kong and used these data to evaluate relationships between forest cover, body size, and rainfall on survival and to estimate population sizes. Hong Kong has been subjected to repeated historic territory-wide deforestation, and thus, we wanted to determine whether there was a link between forest extent as a proxy of habitat quality and newt demography. Annual survival was positively associated with forest cover within core habitat of all populations and negatively related to body size. Mean annual survival (~60%) was similar to that of other stream-dwelling amphibians, but varied among years and declined substantially in 2012-2013, perhaps due to illegal collection. Despite the link between forest extent and survival, population sizes declined at the most forested site by 40% and increased by 104% and 134% at two others. Forest protection and consequential secondary succession during recent decades in Hong Kong may have been responsible for persistence of P. hongkongensis populations.

Nematomorpha from Hong Kong and description of a new species from Malaysia.

We report here for the first time horsehair worms (Nematomorpha) from Hong Kong (China). Most specimens belong to the genus Chordodes, and can be assigned to C. bipilus, C. caledoniensis and to C. cf. moutoni. Two specimens have a cuticular pattern resembling the South American species C. peraccae or C. bouvieri, but for Southeast Asia this pattern is new. These two specimens are reported here as Chordodes sp. Additionally, Gordionus chinensis was found. One new species, Acutogordius koljai, is described from Malaysia. It has a characteristically-structured region posterior to the postcloacal crescent which sets it apart from previously described species in this genus.

Evolutionary response to global change: Climate and land use interact to shape color polymorphism in a woodland salamander.

Evolutionary change has been demonstrated to occur rapidly in human-modified systems, yet understanding how multiple components of global change interact to affect adaptive evolution remains a critical knowledge gap. Climate change is predicted to impose directional selection on traits to reduce thermal stress, but the strength of directional selection may be mediated by changes in the thermal environment driven by land use. We examined how regional climatic conditions and land use interact to affect genetically based color polymorphism in the eastern red-backed salamander (Plethodon cinereus). P. cinereus is a woodland salamander with two primary discrete color morphs (striped, unstriped) that have been associated with macroclimatic conditions. Striped individuals are most common in colder regions, but morph frequencies can be variable within climate zones. We used path analysis to analyze morph frequencies among 238,591 individual salamanders across 1,170 sites in North America. Frequency of striped individuals was positively related to forest cover in populations occurring in warmer regions (>7°C annually), a relationship that was weak to nonexistent in populations located in colder regions (≤7°C annually). Our results suggest that directional selection imposed by climate warming at a regional scale may be amplified by forest loss and suppressed by forest persistence, with a mediating effect of land use that varies geographically. Our work highlights how the complex interaction of selection pressures imposed by different components of global change may lead to divergent evolutionary trajectories among populations.

Diet of the endangered big-headed turtle Platysternon megacephalum.

Populations of the big-headed turtle Platysternon megacephalum are declining at unprecedented rates across most of its distribution in Southeast Asia owing to unsustainable harvest for pet, food, and Chinese medicine markets. Research on Asian freshwater turtles becomes more challenging as populations decline and basic ecological information is needed to inform conservation efforts. We examined fecal samples collected from P. megacephalum in five streams in Hong Kong to quantify the diet, and we compared the germination success of ingested and uningested seeds. Fruits, primarily of Machilus spp., were most frequently consumed, followed by insects, plant matter, crabs and mollusks. The niche breadth of adults was wider than that of juveniles. Diet composition differed between sites, which may be attributable to the history of illegal trapping at some sites, which reduced the proportion of larger and older individuals. Digestion of Machilus spp. fruits by P. megacephalum enhanced germination success of seeds by about 30%. However, most digested seeds are likely defecated in water in this highly aquatic species, which limits the potential benefit to dispersal. The results of our study can be used by conservation-related captive breeding programs to ensure a more optimal diet is provided to captive P. megacephalum.

Is an assessment factor of 10 appropriate to account for the variation in chemical toxicity to freshwater ectotherms under different thermal conditions?

Ecotoxicity tests are often conducted following standard methods, and thus carried out at a fixed water temperature under controlled laboratory conditions. Yet, toxicity of a chemical contaminant may vary in a temperature-dependent manner, depending on the physiology of the test organism and physicochemical properties of the chemical. Although an assessment factor of 10 (AF10) is commonly adopted to account for variability in toxicity data related to temperature in the development of water quality guidelines and/or ecological risk assessment, no one has ever rigorously assessed the appropriateness of AF10 to account for potential variation in temperature-dependent chemical toxicity to aquatic organisms. This study, therefore, aims to address this issue through a meta-analysis by comparing median lethal concentration data for nine chemicals (cadmium, copper, nickel, lead, silver, zinc, arsenic, selenium and DDT) on a range of freshwater ectothermic animal species at different temperatures, and to assess whether AF10 is under- or over-protective for tropical and temperate freshwater ecosystems. Our results reveal varying extents of interaction between temperature and different chemicals on organisms and the complexity of these interactions. Applying AF10 sufficiently protects 90% of the animal species tested over a range of temperatures for cadmium, copper, nickel, silver, zinc and DDT in the tropics, but it is insufficient to adequately encompass a larger temperature variation for most studied chemicals in temperate regions. It is therefore important to set specific AFs for different climatic zones in order to achieve the desired level of ecosystem protection.

Road deicing salt irreversibly disrupts osmoregulation of salamander egg clutches.

It has been postulated that road deicing salts are sufficiently diluted by spring rains to ameliorate any physiological impacts to amphibians breeding in wetlands near roads. We tested this conjecture by exposing clutches of the spotted salamander (Ambystoma maculatum) to three chloride concentrations (1 mg/L, 145 mg/L, 945 mg/L) for nine days, then transferred clutches to control water for nine days, and measured change in mass at three-day intervals. We measured mass change because water uptake by clutches reduces risks to embryos associated with freezing, predation, and disease. Clutches in controls sequestered water asymptotically. Those in the moderate concentrations lost 18% mass initially and regained 14% after transfer to control water. Clutches in high concentration lost 33% mass and then lost an additional 8% after transfer. Our results suggest that spring rains do not ameliorate the effects of deicing salts in wetlands with extremely high chloride concentrations.

Effects of warming conditions in eastern North American forests on red-backed salamander morphology.

Several studies have reported climate-associated changes in phenotypically plastic traits of amphibians, yet it remains unknown whether amphibians can manifest an evolutionary response to global climate change at the rate and magnitude that it is occurring. To assess this issue, we examined temporal change in the morphology of the red-backed salamander (Plethodon cinereus), a small, abundant woodland salamander distributed widely in eastern North America with two distinct morphotypes: striped individuals associated with cooler microclimates and unstriped individuals associated with warmer microclimates. We compiled morph frequencies for 50,960 individual salamanders from 558 sites as recorded in the published literature and in unpublished field notes of herpetologists between 1908 and 2004. We observed that striping probability increased with increasing latitude, longitude, and elevation and decreased (from 80% to 74% range wide) with time. The combined forces of regional climate warming and, particularly, forest disturbance have evidently been sufficient to cause morphological evolution in this amphibian over the last century.