Identification of Turtle-Shell Growth Year Using Hyperspectral Imaging Combined with an Enhanced Spatial-Spectral Attention 3DCNN and a Transformer.

Turtle shell (Chinemys reecesii) is a prized traditional Chinese dietary therapy, and the growth year of turtle shell has a significant impact on its quality attributes. In this study, a hyperspectral imaging (HSI) technique combined with a proposed deep learning (DL) network algorithm was investigated for the objective determination of the growth year of turtle shells. The acquisition of hyperspectral images was carried out in the near-infrared range (948.72-2512.97 nm) from samples spanning five different growth years. To fully exploit the spatial and spectral information while reducing redundancy in hyperspectral data simultaneously, three modules were developed. First, the spectral-spatial attention (SSA) module was developed to better protect the spectral correlation among spectral bands and capture fine-grained spatial information of hyperspectral images. Second, the 3D convolutional neural network (CNN), more suitable for the extracted 3D feature map, was employed to facilitate the joint spatial-spectral feature representation. Thirdly, to overcome the constraints of convolution kernels as well as better capture long-range correlation between spectral bands, the transformer encoder (TE) module was further designed. These modules were harmoniously orchestrated, driven by the need to effectively leverage both spatial and spectral information within hyperspectral data. They collectively enhance the model's capacity to extract joint spatial and spectral features to discern growth years accurately. Experimental studies demonstrated that the proposed model (named SSA-3DTE) achieved superior classification accuracy, with 98.94% on average for five-category classification, outperforming traditional machine learning methods using only spectral information and representative deep learning methods. Also, ablation experiments confirmed the effectiveness of each module to improve performance. The encouraging results of this study revealed the potentiality of HSI combined with the DL algorithm as an efficient and non-destructive method for the quality control of turtle shells.

Growing as slow as a turtle: Unexpected maturational differences in a small, long-lived species.

Turtle body size is associated with demographic and other traits like mating success, reproductive output, maturity, and survival. As such, growth analyses are valuable for testing life history theory, demographic modeling, and conservation planning. Two important but unsettled research areas relate to growth after maturity and growth rate variation. If individuals exhibit indeterminate growth after maturity, older adults may have an advantage in fecundity, survival, or both over younger/smaller adults. Similarly, depending on how growth varies, a portion of the population may mature earlier, grow larger, or both. We used 23-years of capture-mark-recapture data to study growth and maturity in the Spotted Turtle (Clemmys guttata), a species suffering severe population declines and for which demographic data are needed for development of effective conservation and management strategies. There was strong support for models incorporating sex as a factor, with the interval growth model reparametrized for capture-mark-recapture data producing later mean maturation estimates than the age-based growth model. We found most individuals (94%) continued growing after maturity, but the instantaneous relative annual plastral growth rate was low. We recommend future studies examine the possible contribution of such slow, continued adult growth to fecundity and survival. Even seemingly negligible amounts of annual adult growth can have demographic consequences affecting the population vital rates for long-lived species.

Global Analysis of Transcriptome and Translatome Revealed That Coordinated WNT and FGF Regulate the Carapacial Ridge Development of Chinese Soft-Shell Turtle.

The turtle carapace is composed of severely deformed fused dorsal vertebrae, ribs, and bone plates. In particular, the lateral growth in the superficial layer of turtle ribs in the dorsal trunk causes an encapsulation of the scapula and pelvis. The recent study suggested that the carapacial ridge (CR) is a new model of epithelial-mesenchymal transition which is essential for the arrangement of the ribs. Therefore, it is necessary to explore the regulatory mechanism of carapacial ridge development to analyze the formation of the turtle shell. However, the current understanding of the regulatory network underlying turtle carapacial ridge development is poor due to the lack of both systematic gene screening at different carapacial ridge development stages and gene function verification studies. In this study, we obtained genome-wide gene transcription and gene translation profiles using RNA sequencing and ribosome nascent-chain complex mRNA sequencing from carapacial ridge tissues of Chinese soft-shell turtle at different development stages. A correlation analysis of the transcriptome and translatome revealed that there were 129, 670, and 135 codifferentially expressed genes, including homodirection and opposite-direction differentially expressed genes, among three comparison groups, respectively. The pathway enrichment analysis of codifferentially expressed genes from the Kyoto Encyclopedia of Genes and Genomes showed dynamic changes in signaling pathways involved in carapacial ridge development. Especially, the results revealed that the Wnt signaling pathway and MAPK signaling pathway may play important roles in turtle carapacial ridge development. In addition, Wnt and Fgf were expressed during the carapacial ridge development. Furthermore, we discovered that Wnt5a regulated carapacial ridge development through the Wnt5a/JNK pathway. Therefore, our studies uncover that the morphogenesis of the turtle carapace might function through the co-operation between conserved WNT and FGF signaling pathways. Consequently, our findings revealed the dynamic signaling pathways acting on the carapacial ridge development of Chinese soft-shell turtle and provided new insights into uncover the molecular mechanism underlying turtle shell morphogenesis.

Divergent incubation temperature effects on thermal sensitivity of hatchling performance in two different latitudinal populations of an invasive turtle.

The incubation temperature for embryonic development affects several aspects of hatchling performance, but its impact on the thermal sensitivity of performance attributes remains poorly investigated. In the present study, Trachemys scripta elegans hatchlings from two different latitudinal populations were collected to assess the effects of different incubation temperatures on the locomotor (swimming speed) and physiological (heart rate) performances, and the thermal sensitivity of these two attributes. The incubation temperature significantly affected the examined physiological traits. Hatchling turtles produced at low incubation temperature exhibited relatively higher cold tolerance (lower body temperatures at which the animals lose the ability to escape from the lethal conditions), and reduced heart rate and swimming speed. Furthermore, the effect of incubation temperature on the thermal sensitivity of swimming speed differed between the low- and high-latitude populations. At relatively high incubation temperatures, the high-latitude hatchling turtles exhibited reduced thermal sensitivities of swimming speed than those of the low-latitude ones. Reduced thermal sensitivity of locomotor performance together with high cold tolerance, exhibited by the high-latitude hatchling turtles potentially reflected local adaptation to relatively colder and more thermally-variable environments.

Influence of Herbicide Exposure and Ranavirus Infection on Growth and Survival of Juvenile Red-Eared Slider Turtles (Trachemys scripta elegans).

Ranaviruses are an important wildlife pathogen of fish, amphibians, and reptiles. Previous studies have shown that susceptibility and severity of infection can vary with age, host species, virus strain, temperature, population density, and presence of environmental stressors. Experiments are limited with respect to interactions between this pathogen and environmental stressors in reptiles. In this study, we exposed hatchling red-eared slider turtles (Trachemys scripta elegans) to herbicide and ranavirus treatments to examine direct effects and interactions on growth, morbidity, and mortality. Turtles were assigned to one of three herbicide treatments or a control group. Turtles were exposed to atrazine, Roundup ProMax®, or Rodeo® via water bath during the first 3 weeks of the experiment. After 1 week, turtles were exposed to either a control (cell culture medium) or ranavirus-infected cell lysate via injection into the pectoral muscles. Necropsies were performed upon death or upon euthanasia after 5 weeks. Tissues were collected for histopathology and detection of ranavirus DNA via quantitative PCR. Only 57.5% of turtles exposed to ranavirus tested positive for ranaviral DNA at the time of death. Turtles exposed to ranavirus died sooner and lost more mass and carapace length, but not plastron length, than did controls. Exposure to environmentally relevant concentrations of herbicides did not impact infection rate, morbidity, or mortality of hatchling turtles due to ranavirus exposure. We also found no direct effects of herbicide or interactions with ranavirus exposure on growth or survival time. Results of this study should be interpreted in the context of the modest ranavirus infection rate achieved, the general lack of growth, and the unplanned presence of an additional pathogen in our study.

Influence of incubation temperature on embryo development, hatchling morphology and early growth rate in red-footed tortoise (Chelonoidis carbonaria).

Reptile embryos respond to temperature changes with metabolic and physiological adjustments that influence hatchling success, phenotype, behaviour, and growth rate. Climate change and global warming can affect the reptile population by altering the frequencies of hatchling survival and phenotypes. Therefore, previous studies proposed artificial incubation as a potential strategy for mitigating these effects. Red-footed tortoise (Chelonoidis carbonaria) eggs were collected and incubated at constant temperatures of 27.5 °C and 29.5 °C to investigate the physiological effects of temperature on embryo development, hatchling morphology, and early post-hatch growth rate. The direct effect of temperature on the incubation period, egg mass loss, hatching success, hatchling size, and mass was evaluated at hatching and three months of age. Hatchlings from 29.5 °C presented a shorter incubation period (141 days) than those from 27.5 °C (201 days; p < 0.05). Egg mass loss, hatchling mass, and size at hatching were not different between the incubation temperatures (p > 0.05). However, the hatching success (survival rate) was lower (64.5% versus 100%) in eggs incubated at 29.5 °C, but the hatchling mass and straight plastron width were higher at three months of age than those from eggs incubated at 27.5 °C (p < 0.05). These results indicate that incubation temperature influences hatching success and hatchling size and mass in the first months by influencing the early growth rate.

Housing experience affects adult neurogenesis in turtles (Chrysemys picta).

Adult neurogenesis increases in mammals when they are exposed to an enriched environment or given the opportunity to exercise. In this experiment, we investigated whether turtles would show differences in the number of new neurons in the telencephalon when they were exposed to deep water, conspecifics, and plants and logs (EE group), compared to a group of animals housed in individual cages with shallow water (IN group). A control group (EX) was given deep water and conspecifics but no plants and logs. We gave nine injections of BrdU over a 3-week period, starting when the turtles were introduced to the housing. The results showed that both the EE and the EX groups had more new cells in the dorsal ventricular ridge (DVR), a sensory area of the telencephalon. The two groups did not differ from one another. The group-housed animals also had a higher percentage of new neurons in the DVR that were double labeled for NeuN, a marker of neurons, compared to the IN group. There were no significant differences between groups in the number of new cells in the medial cortex, the homolog of the hippocampus. These findings demonstrate that the housing experience influences the number of new cells that survive in the brains of turtles. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Nontraditional systems in aging research: an update.

Research on the evolutionary and mechanistic aspects of aging and longevity has a reductionist nature, as the majority of knowledge originates from experiments on a relatively small number of systems and species. Good examples are the studies on the cellular, molecular, and genetic attributes of aging (senescence) that are primarily based on a narrow group of somatic cells, especially fibroblasts. Research on aging and/or longevity at the organismal level is dominated, in turn, by experiments on Drosophila melanogaster, worms (Caenorhabditis elegans), yeast (Saccharomyces cerevisiae), and higher organisms such as mice and humans. Other systems of aging, though numerous, constitute the minority. In this review, we collected and discussed a plethora of up-to-date findings about studies of aging, longevity, and sometimes even immortality in several valuable but less frequently used systems, including bacteria (Caulobacter crescentus, Escherichia coli), invertebrates (Turritopsis dohrnii, Hydra sp., Arctica islandica), fishes (Nothobranchius sp., Greenland shark), reptiles (giant tortoise), mammals (blind mole rats, naked mole rats, bats, elephants, killer whale), and even 3D organoids, to prove that they offer biogerontologists as much as the more conventional tools. At the same time, the diversified knowledge gained owing to research on those species may help to reconsider aging from a broader perspective, which should translate into a better understanding of this tremendously complex and clearly system-specific phenomenon.

Neonatal biometry and characterization of Amazonian Turtle nests (Podocnemis expansa)

The present study aimed to study the reproduction of the Amazon turtle (Podocnemis expansa) through the evaluation of the reproductive parameters of adult females. This study was carried out in the Crixás-Açu River, municipality of Mundo Novo, state of Goiás, Brazil. In September, the biometrics of 20 females were evaluated during the spawning period. Their nests were marked for subsequent evaluation of hatchlings, measuring the distances of each nest to the river and vegetation. The second stage consisted of the evaluation of hatchlings and characterization of nests after egg hatching. The datawere tested using the Pearson Correlation to measure the degree of linear correlation between the variables analyzed, such as the parameters of females, hatchlings, nests, and eggs. Nest depth was positively correlated with the number of eggshells found. We also found a correlation between the number of eggsshells and the distance of nests to the vegetation. Although moderate, this result indicates that as the distance to the vegetation increases, the number of eggshells found in nests decreases. The spawning site for Amazon turtle breeding is crucial to determine a higher number of eggshatching and better development of hatchlings. However, the literature is still very scarce regarding the choice of spawning grounds and the influence that vegetation may have on the development of freshwater turtle hatchlingsin Brazil.

Long-term survey of sea turtles (Caretta caretta) reveals correlations between parasite infection, feeding ecology, reproductive success and population dynamics.

Long-term monitoring of host-parasite interactions is important for understanding the consequences of infection on host fitness and population dynamics. In an eight-year survey of the loggerhead sea turtle (Caretta caretta) population nesting in Cabo Verde, we determined the spatiotemporal variation of Ozobranchus margoi, a sanguivorous leech best known as a vector for sea turtle fibropapilloma virus. We quantified O. margoi association with turtles' δ15N and δ13C stable isotopes to identify where infection occurs. We then measured the influence of infection on reproduction and offspring fitness. We found that parasite prevalence has increased from 10% of the population in 2010, to 33% in 2017. Stable isotope analysis of host skin samples suggests transmission occurs within the host's feeding grounds. Interestingly, we found a significant interaction between individual size and infection on the reproductive success of turtles. Specifically, small, infected females produced fewer offspring of poorer condition, while in contrast, large, infected turtles produced greater clutch sizes and larger offspring. We interpret this interaction as evidence, upon infection, for a size-dependent shift in reproductive strategy from bet hedging to terminal investment, altering population dynamics. This link between infection and reproduction underscores the importance of using long-term monitoring to quantify the impact of disease dynamics over time.

Effect of acclimation temperature and substrate type on selected temperature, movement and activity of juvenile spiny softshell turtles (Apalone spinifera) in an aquatic thermal gradient.

In some turtle species, temperature selection may be influenced by environmental conditions, including acclimation temperature and substrate quality. These factors may be particularly important for softshell turtles that are highly aquatic and often thermoregulate by burying in the substrate in shallow water microhabitats. We tested for effects of acclimation temperature (22 °C or 27 °C) and substrate type (sand or gravel) on the selected temperature and movement patterns of 20 juvenile spiny softhshell turtles (Apalone spinifera; Reptilia: Trionychidae) in an aquatic thermal gradient of 14-34 °C. Among 7-11 month old juvenile softshell turtles, acclimation temperature and substrate type did not influence temperature selection, nor alter activity and movement patterns. During thermal gradient tests, both 22- and 27 °C-acclimated turtles selected the warmest temperature (34 °C) available most frequently, regardless of substrate type (sand or gravel). Similarly, acclimation temperature and substrate type did not influence movement patterns of turtles, nor the number of chambers used in the gradient tests. These results suggest that juvenile Apalone spinifera are capable of detecting small temperature increments and prefer warm temperatures that may positively influence growth and metabolism, and that thermal factors more significantly influence aquatic thermoregulation in this species than does substrate type.

Identifying patterns in foraging-area origins in breeding aggregations of migratory species: Loggerhead turtles in the Northwest Atlantic.

Population assessments conducted at reproductive sites of migratory species necessitate understanding the foraging-area origins of breeding individuals. Without this information, efforts to contextualize changes in breeding populations and develop effective management strategies are compromised. We used stable isotope analysis of tissue samples collected from loggerhead sea turtles (Caretta caretta) nesting at seven sites in the Northern Recovery Unit (NRU) of the eastern United States (North Carolina, South Carolina and Georgia) to assign females to three separate foraging areas in the Northwest Atlantic Ocean (NWA). We found that the majority of the females at NRU nesting sites (84.4%) use more northern foraging areas in the Mid-Atlantic Bight, while fewer females use more proximate foraging areas in the South Atlantic Bight (13.4%) and more southerly foraging areas in the Subtropical Northwest Atlantic (2.2%). We did not find significant latitudinal or temporal trends in the proportions of NRU females originating from different foraging areas. Combining these findings with previous data from stable isotope and satellite tracking studies across NWA nesting sites showed that variation in the proportion of adult loggerheads originating from different foraging areas is primarily related differences between recovery units: individuals in the NRU primarily use the Mid-Atlantic Bight foraging area, while individuals from the three Florida recovery units primarily use the Subtropical Northwest Atlantic and Eastern Gulf of Mexico foraging areas. Because each foraging area is associated with its own distinct ecological characteristics, environmental fluctuations and anthropogenic threats that affect the abundance and productivity of individuals at nesting sites, this information is critical for accurately evaluating population trends and developing effective region-specific management strategies.

Seascape Genetics and the Spatial Ecology of Juvenile Green Turtles.

Understanding how ocean currents impact the distribution and connectivity of marine species, provides vital information for the effective conservation management of migratory marine animals. Here, we used a combination of molecular genetics and ocean drift simulations to investigate the spatial ecology of juvenile green turtle (Chelonia mydas) developmental habitats, and assess the role of ocean currents in driving the dispersal of green turtle hatchlings. We analyzed mitochondrial (mt)DNA sequenced from 358 juvenile green turtles, and from eight developmental areas located throughout the Southwest Indian Ocean (SWIO). A mixed stock analysis (MSA) was applied to estimate the level of connectivity between developmental sites and published genetic data from 38 known genetic stocks. The MSA showed that the juvenile turtles at all sites originated almost exclusively from the three known SWIO stocks, with a clear shift in stock contributions between sites in the South and Central Areas. The results from the genetic analysis could largely be explained by regional current patterns, as shown by the results of passive numerical drift simulations linking breeding sites to developmental areas utilized by juvenile green turtles. Integrating genetic and oceanographic data helps researchers to better understand how marine species interact with ocean currents at different stages of their lifecycle, and provides the scientific basis for effective conservation management.

Population dynamics and biological feasibility of sustainable harvesting as a conservation strategy for tropical and temperate freshwater turtles.

BACKGROUND: Conservation strategies are urgently needed for tropical turtles that are increasingly threatened by unsustainable exploitation. Studies conducted exclusively in temperate zones have revealed that typical turtle life history traits (including delayed sexual maturity and high adult survivorship) make sustainable harvest programs an unviable strategy for turtle conservation. However, most turtles are tropical in distribution and the tropics have higher, more constant and more extended ambient temperature regimes that, in general, are more favorable for population growth. METHODS: To estimate the capacity of temperate and tropical turtles to sustain harvest, we synthesized life-history traits from 165 predominantly freshwater turtle species in 12 families (Carettochelydae, Chelidae, Chelydridae, Dermatemydidae, Emydidae, Geoemydidae, Kinosternidae, Pelomedusidae, Platysternidae, Podocnemididae, Staurotypidae and Trionychidae). The influence of climate variables and latitude on turtle life-history traits (clutch size, clutch frequency, age at sexual maturity, and annual adult survival) were examined using Generalized Additive Models. The biological feasibility of sustainable harvest in temperate and tropical species was evaluated using a sensitivity analysis of population growth rates obtained from stage-structured matrix population models. RESULTS: Turtles at low latitudes (tropical zones) exhibit smaller clutch sizes, higher clutch frequency, and earlier age at sexual maturity than those at high latitudes (temperate zones). Adult survival increased weakly with latitude and declined significantly with increasing bioclimatic temperature (mean temperature of warmest quarter). A modeling synthesis of these data indicates that the interplay of life-history traits does not create higher harvest opportunity in adults of tropical species. Yet, we found potential for sustainable exploitation of eggs in tropical species. CONCLUSIONS: Sustainable harvest as a conservation strategy for tropical turtles appears to be as biologically problematic as in temperature zones and likely only possible if the focus is on limited harvest of eggs. Further studies are urgently needed to understand how the predicted population surplus in early life stages can be most effectively incorporated into conservation programs for tropical turtles.

Contribution of green turtles Chelonia mydas to total herbivore biomass in shallow tropical reefs of oceanic islands.

Green turtles are megaherbivores with a key role in the dynamics of tropical seagrass meadows, but little is known about their relevance as herbivores in tropical reef habitats. We conducted underwater censuses of green turtles, herbivorous fishes and sea urchins in two distinct tropical regions: Fernando de Noronha (Western Atlantic Ocean) and the Hawaiian Archipelago (Central Pacific Ocean), to assess the contribution of green turtles to the total herbivore biomass in shallow reef habitats of tropical oceanic islands. Juvenile green turtles ranging 40-60 cm were observed at most of the surveyed sites, and hence, could be considered typical components of the shallow reef fauna of tropical oceanic islands. Furthermore, they were usually one of the most abundant species of roving herbivores in many of the sites surveyed. However, the biomass of green turtles was usually much lower than the aggregated biomass of fishes or sea urchins, which usually constituted most of the total herbivore biomass. Green turtles made a major contribution to the total herbivore biomass only in sheltered sites with low rugosity, low coral cover and high algal cover. Further investigation on the trophic redundancy between herbivores is required to assess the actual relevance of green turtles in reef ecosystems of oceanic islands, compared to herbivorous fishes and sea urchins, because different herbivores may target different algal resources and complementarity may be needed to maintain ecosystem functioning across large, naturally varied reefscapes.

Development-specific transcriptomic profiling suggests new mechanisms for anoxic survival in the ventricle of overwintering turtles.

Oxygen deprivation swiftly damages tissues in most animals, yet some species show remarkable abilities to tolerate little or even no oxygen. Painted turtles exhibit a development-dependent tolerance that allows adults to survive anoxia approximately four times longer than hatchlings: adults survive ∼170 days and hatchlings survive ∼40 days at 3°C. We hypothesized that this difference is related to development-dependent differences in ventricular gene expression. Using a comparative ontogenetic approach, we examined whole transcriptomic changes before, during and 5 days after a 20-day bout of anoxic submergence at 3°C. Ontogeny accounted for more gene expression differences than treatment (anoxia or recovery): 1175 versus 237 genes, respectively. Of the 237 differences, 93 could confer protection against anoxia and reperfusion injury, 68 could be injurious and 20 may be constitutively protective. Most striking during anoxia was the main expression pattern of all 76 annotated ribosomal protein (R-protein) mRNAs, which decreased in anoxia-tolerant adults, but increased in anoxia-sensitive hatchlings, suggesting adult-specific regulation of translational suppression. These genes, along with 60 others that decreased their levels in adults and either increased or remained unchanged in hatchlings, implicate antagonistic pleiotropy as a mechanism to resolve the long-standing question about why hatchling painted turtles overwinter in terrestrial nests, rather than emerge and overwinter in water during their first year. In summary, developmental differences in the transcriptome of the turtle ventricle revealed potentially protective mechanisms that contribute to extraordinary adult-specific anoxia tolerance, and provide a unique perspective on differences between the anoxia-induced molecular responses of anoxia-tolerant and anoxia-sensitive phenotypes within a species.

Population recovery changes population composition at a major southern Caribbean juvenile developmental habitat for the green turtle, Chelonia mydas.

Understanding the population composition and dynamics of migratory megafauna at key developmental habitats is critical for conservation and management. The present study investigated whether differential recovery of Caribbean green turtle (Chelonia mydas) rookeries influenced population composition at a major juvenile feeding ground in the southern Caribbean (Lac Bay, Bonaire, Caribbean Netherlands) using genetic and demographic analyses. Genetic divergence indicated a strong temporal shift in population composition between 2006-2007 and 2015-2016 (ϕST = 0.101, P < 0.001). Juvenile recruitment (<75.0 cm straight carapace length; SCL) from the north-western Caribbean increased from 12% to 38% while recruitment from the eastern Caribbean region decreased from 46% to 20% between 2006-2007 and 2015-2016. Furthermore, the product of the population growth rate and adult female abundance was a significant predictor for population composition in 2015-2016. Our results may reflect early warning signals of declining reproductive output at eastern Caribbean rookeries, potential displacement effects of smaller rookeries by larger rookeries, and advocate for genetic monitoring as a useful method for monitoring trends in juvenile megafauna. Furthermore, these findings underline the need for adequate conservation of juvenile developmental habitats and a deeper understanding of the interactions between megafaunal population dynamics in different habitats.