Resilience of marine turtle regional management units to climate change

Enhancing species resilience to changing environmental conditions is often suggested as a climate change adaptation strategy. To effectively achieve this, it is necessary first to understand the factors that determine species resilience, and their relative importance in shaping the ability of species to adjust to the complexities of environmental change. This is an extremely challenging task because it requires comprehensive information on species traits. We explored the resilience of 58 marine turtle regional management units (RMUs) to climate change, encompassing all seven species of marine turtles worldwide. We used expert opinion from the IUCN‐SSC Marine Turtle Specialist Group (n = 33 respondents) to develop a Resilience Index, which considered qualitative characteristics of each RMU (relative population size, rookery vulnerability, and genetic diversity) and non climate‐related threats (fisheries, take, coastal development, and pollution/pathogens). Our expert panel perceived rookery vulnerability (the likelihood of functional rookeries becoming extirpated) and non climate‐related threats as having the greatest influence on resilience of RMUs to climate change. We identified the world's 13 least resilient marine turtle RMUs to climate change, which are distributed within all three major ocean basins and include six of the world's seven species of marine turtle. Our study provides the first look at inter‐ and intra‐species variation in resilience to climate change and highlights the need to devise metrics that measure resilience directly. We suggest that this approach can be widely used to help prioritize future actions that increase species resilience to climate change.     

Efficient embryonic culture method for the Japanese striped snake,Elaphe quadrivirgata,and its early developmental stages

The morphogenesis of snake embryos is an elusive yet fascinating research target for developmental biologists. However, few data exist on development of early snake embryo due to limited availability of pregnant snakes, and the need to harvest early stage embryos directly from pregnant snakes before oviposition without knowing the date of fertilization. We established an ex vivo culture method for early snake embryos using the Japanese striped snake, Elaphe quadrivirgata. This method, which we named “sausage‐style (SS) culture”, allows us to harvest snake embryos at specific stages for each experiment. Using this SS culture system, we calculated somite formation rate at early stages before oviposition. The average somite formation rate between 6/7 and 12/13 somite stages was 145.9 min, between 60/70 and 80/91 somite stages 42.4 min, and between 113–115 and 126/127 somite stages 71 min. Thus, somite formation rate that we observed during early snake embryogenesis was changed over time. We also describe a developmental staging series for E. quadrivirgata. This is the first report of a developmental series of early snake embryogenesis prior to oviposition by full‐color images with high‐resolution. We propose that the SS culture system is an easy method for treating early snake embryos ex vivo.     

Embryogenic staging of fugu, Takifugu rubripes, and expression profiles of aldh1a2, aldh1a3 and cyp26a1

Fugu (Takifugu rubripes) has contributed as an ideal model organism for understanding the structure and evolution of vertebrate genomes, but also has potential as a good model organism for developmental biology because of the availability of the genome information. However, there is no comprehensive report describing the developmental stages, which is fundamental data for developmental biology. Here we describe a series of stages of the embryonic development of fugu during the first 8 days after fertilization, i.e. from fertilization to hatching. We define seven periods of embryogenesis – the zygote, cleavage, blastula, gastrula, segmentation, pharyngula, and hatching periods. Stages subdividing these periods are defined based on morphological characteristics. In addition, as a model experiment of gene expression analysis using this staging series, we performed in situ hybridization of aldh1a2, aldh1a3 and cyp26a1 that play regulatory roles in retinoic acid (RA) metabolism essential for embryogenesis. This report provides fundamental information on fugu embryogenesis, which is anticipated to facilitate the use of fugu as a model organism for developmental studies.     

Post-embryonic development of sheepshead minnow <Emphasis Type="Italic">Cyprinodon variegatus</Emphasis>: a staging tool based on externally visible anatomical traits

The sheepshead minnow Cyprinodon variegatus has become a favoured model for laboratory studies because of their small size, rapid development, and tolerance of laboratory conditions. Here, we analyse sheepshead minnow post-embryonic development with the goal of providing a generally useful method for staging fish after embryogenesis. Groups of three females and two males were placed in breeding chambers designed for this experiment. More than 100 eggs were collected and maintained in seawater. Embryos were selected under a dissection microscope and placed in incubation dishes (50 per dish) at 26 °C. On day six, embryos hatched and larvae were transferred to 1 L beakers. To define a simplified normalization table for sheepshead minnow development, we measured each fish for its standard length and examined the fish for four externally evident traits: pigmentation pattern, caudal fin morphology, anal fin morphology, and dorsal fin morphology. The four traits were chosen, because they are easily visualized with standard laboratory equipment such as the stereomicroscope and camera. We have provided criteria for staging sheepshead minnows in studies of post-embryonic development. Our data suggest that dorsal and anal fin morphology may serve as a useful phenotype for defining metamorphic climax stages throughout post-embryonic development in C. variegatus. The staging systems we propose should facilitate detailed anatomical and developmental analyses in relation to ecotoxicological studies on potential disruption of the thyroid axis by xenobiotics and endocrine-disrupting compounds.     

Morphological and temporal variation in early embryogenesis contributes to species divergence in Malawi cichlid fishes

The cichlid fishes comprise the largest extant vertebrate family and are the quintessential example of rapid “explosive” adaptive radiations and phenotypic diversification. Despite low genetic divergence, East African cichlids harbor a spectacular intra- and interspecific morphological diversity, including the hyper-variable, neural crest (NC)-derived traits such as coloration and craniofacial skeleton. Although the genetic and developmental basis of these phenotypes has been investigated, understanding of when, and specifically how early, in ontogeny species-specific differences emerge, remains limited. Since adult traits often originate during embryonic development, the processes of embryogenesis could serve as a potential source of species-specific variation. Consequently, we designed a staging system by which we compare the features of embryogenesis between three Malawi cichlid species—Astatotilapia calliptera, Tropheops sp. ‘mauve’ and Rhamphochromis sp. “chilingali”—representing a wide spectrum of variation in pigmentation and craniofacial morphologies. Our results showed fundamental differences in multiple aspects of embryogenesis that could underlie interspecific divergence in adult adaptive traits. First, we identified variation in the somite number and signatures of temporal variation, or heterochrony, in the rates of somite formation. The heterochrony was also evident within and between species throughout ontogeny, up to the juvenile stages. Finally, the identified interspecific differences in the development of pigmentation and craniofacial cartilages, present at the earliest stages of their overt formation, provide compelling evidence that the species-specific trajectories begin divergence during early embryogenesis, potentially during somitogenesis and NC development. Altogether, our results expand our understanding of fundamental cichlid biology and provide new insights into the developmental origins of vertebrate morphological diversity.     

The ontogeny of color: developmental origins of divergent pigmentation in Drosophila americana and D. novamexicana

Pigmentation is a model trait for evolutionary and developmental analysis that is particularly amenable to molecular investigation in the genus Drosophila. To better understand how this phenotype evolves, we examined divergent pigmentation and gene expression over developmental time in the dark‐bodied D. americana and its light‐bodied sister species D. novamexicana. Prior genetic analysis implicated two enzyme‐encoding genes, tan and ebony, in pigmentation divergence between these species, but questions remain about the underlying molecular mechanisms. Here, we describe stages of pupal development in both species and use this staging to determine when pigmentation develops and diverges between D. americana and D. novamexicana. For the developmental stages encompassing pigment divergence, we compare mRNA expression of tan and ebony over time and between species. Finally, we use allele‐specific expression assays to determine whether interspecific differences in mRNA abundance have a cis‐regulatory basis and find evidence of cis‐regulatory divergence for both tan and ebony. cis‐regulatory divergence affecting tan had a small effect on mRNA abundance and was limited to a few developmental stages, yet previous data suggests that this divergence is likely to be biologically meaningful. Our study suggests that small and developmentally transient expression changes may contribute to phenotypic diversification more often than commonly appreciated. Recognizing the potential phenotypic impact of such changes is important for a scientific community increasingly focused on dissecting quantitative variation, but detecting these types of changes will be a major challenge to elucidating the molecular basis of complex traits.     

Resources available to individuals and organisations involved with marine turtle research and conservation in the Mediterranean

Abstract

We review some key resources freely or cheaply available to those involved with marine turtle research and conservation in the Mediterranean. These include: Marine Turtle Newsletter, Listservs, bibliographic databases, Mediterranean specific technical manuals, IUCN Marine Turtle Specialist Group publications, internet sources and books.     

Adult and early‐stage characters of Brassolini contain conflicting phylogenetic signal (Lepidoptera,Nymphalidae)

This study examines the contribution of early‐stages and adult characters to the reconstruction of the phylogeny of Brassolini butterflies. Parsimony analyses used both equal weights and implied weights, and a series of analyses were performed. First, we analysed adult and early‐stages partitions independently and in combination for a subset of 27 species; in these cases the matrices were mostly complete. Whereas the adult partition alone produced a topology that was well resolved and congruent with previous studies, the early‐stages partition produced a poorly resolved tree under equal weights. Furthermore, implied weights produced a well‐resolved early‐stages topology that differed significantly from the adult topology. When both partitions were combined for 27 species, implied weights yielded a topology that resembled the adult tree except for the positions of Bia and Penetes, but statistical node support was generally lower. This suggests that stochastic noise increased when early‐stage characters were added to the adult partition, but the combined partitions topology was not statistically different from that based on adult characters alone. Second, given that preserved early stages are not as readily available as adults, we analysed a matrix including 45 species in which early‐stage data were missing for 18 species, and compared the topology to that produced by the adult partition alone. Results were similar to the analyses including fewer species; the combined partitions tree was similar to that from the adult partition except for the position of Bia and Penetes. We compare our findings to other genus‐level phylogenetic studies within Lepidoptera that have also used early‐stages and adult characters.     

Effects of habitat edges and nest‐site characteristics on Painted Bunting nest success

Declines in populations of Painted Buntings (Passerina ciris) over the past several decades have led to their recent classification as a species of conservation concern. To better assess their status, we investigated factors associated with productivity and abundance of a population in south‐central Louisiana during 2010–2011. We monitored 41 Painted Bunting nests, 14 with video cameras, to identify predators, parasitism events, and improve nest success estimates. Vegetation measurements were also collected at nest sites and non‐nest sites to quantify habitat characteristics. We used an information‐theoretic approach to evaluate support for multiple models evaluating nest success. Highly supported models indicated large negative effects of Brown‐headed Cowbird (Molothrus ater) parasitism, and positive effects of increased canopy cover and distance to habitat edge on daily survival rates. Our estimate of daily survival rate was 0.94 ± 0.03 and the probability of survival was 0.25 ± 0.02. Point‐count data revealed that densities of Painted Buntings were greater in treeline habitats than in open scrub‐shrub and mature forest edge habitats. Furthermore, treelines had higher densities of large trees (>23 cm dbh) and percent canopy cover, variables positively associated with nest success, than open scrub‐shrub and forest edge habitats. In general, survival rates and causes of nest failure in southern Louisiana were similar to those determined for breeding populations of Painted Buntings at other sites. Our results suggest that treelines, despite having a high edge‐to‐area ratio, might be preferred nesting habitat for Painted Buntings in our study area.     

Spatial distribution of turtle barnacles on the green sea turtle, Chelonia mydas

Distribution patterns of epibiotic barnacles on green sea turtles were investigated in waters neighboring Okinawa, Japan. A number of barnacle species were found to coexist on the turtles and were classified into three genera: Chelonibia, Platylepas and Stomatolepas. Attachment sites on the turtles varied among the barnacle species, suggesting that there is niche partitioning with respect to their microhabitat selection. Turtle bodies offer a “patchy” environment for barnacles, so we also analyzed coexistence patterns in the context of an aggregation model. Within each genus, individual barnacles showed a clumped distribution. The different genera do not have mutually exclusive distribution patterns, but instead occur on the same turtle to various degrees. However, when turtles were divided into two size classes, both the level of aggregation and the degree of interspecific overlap among the barnacles was significantly lower on large turtles. We suggest that obtaining basic information on turtle epibionts will shed light on the biology of wild turtles, which is still largely unknown.     

Catching the phylogenic history through the ontogenic hourglass: a phylogenomic analysis of Drosophila body segmentation genes

SUMMARY The phylogenetic information content of different developmental stages is a long‐standing issue in the study of development and evolution. We performed phylogenetic analyses of 51 body segmentation genes in 12 species of Drosophila in order to investigate the impact of the mode of evolution of development on phylogeny inference. Previous studies of these genes in Drosophila using pairwise phenetic comparisons at the species group level revealed the presence of an “hourglass model” (HG), wherein mid‐embryonic stages are the most evolutionarily constrained. We utilized two character‐based approaches: taxonomic congruence using the relative consensus fork index (RCFI), in which phylogenies are inferred from each gene separately and compared with a total evidence tree (TET), and partitioned simultaneous analysis using several indices such as branch support (BS) and localized incongruence length difference (LILD) test. We also proposed a new index, the recapitulatory index (R), which divides the number of synapomorphies on the total number of informative characters in a data set. Polynomial adjustment of both BS and R indices showed strong support for the hourglass model regardless of the taxonomic level (species subgroup vs. subgenera), showing less phylogenetic information content for mid‐developmental stages (mainly the zygotic segment polarity stage). Significant LILD scores were randomly distributed among developmental stages revealing the absence of differential selective constraints, but were significantly related to chromosomal location showing physical (linkage) impact on phylogenetic incongruence. RCFI was the most sensitive measure to taxonomic level, having a convex parabola at the species subgroup level in support of the hourglass model and a concave parabola at the subgeneric level in support of the adaptive penetrance model. This time‐dependent discrepancy of best fit developmental model parallels previous conflicting results from the vertebrates. Because of the quasi‐phenetic nature of this index, we argue that the discrepancy is due to the evolutionary rate heterogeneity of developmental genes rather than to fundamental differences among organisms. We suggest that simultaneous character‐based analyses give better macroevolutionary support to the hourglass model of the developmental constraints on genome evolution than pairwise phenetic comparisons.     

Developmental staging table of the green iguana

Embryonic staging tables provide information to standardize embryological investigations and to subsidize discussions about evolution. We have established a developmental staging table for Iguana iguana iguana. The sample was composed of 142 embryos, incubated at a constant temperature and collected at regular intervals. Morphological features as pharyngeal arches, craniofacial structures, eyes, limbs, claws, pigmentation, scales and egg tooth were evaluated to determine development stages. The normal staging table includes 17 stages from oviposition to hatching, based on chronology and morphological external features. Stages from 1 to 27 occur before oviposition. Stage 28 was the first described, because all embryos presented limb bud anlage, key feature of the previous stage. We used pharyngeal arches and limb buds to describe the first stages; claws, genital papilla and scales to describe the middle stages; and pigmentation, size and egg tooth to describe the last stages. Incubation lasted approximately 2 months in a controlled environment. The results were similar to the data from other lizards, confirming the embryonic conservative pattern of the group.     

Effects of biotic and abiotic factors on the oxygen content of green sea turtle nests during embryogenesis

Several biotic and abiotic factors can influence nest oxygen content during embryogenesis. Several of these factors were determined during each developmental stage of green sea turtle embryos on Wan-an Island, Penghu Archipelago, Taiwan. We examined oxygen content in 7 nests in 2007 and 11 in 2008. Oxygen in the adjacent sand, total and viable clutch sizes, air, sand and nest temperatures, and sand characters of each nest were also determined. Oxygen content was lower in late stages than in the early and middle stages. It was also lower in the middle layer than in the upper and bottom layers. Nest temperature showed opposite trends, reaching its maximum value in late stages of development. Nest oxygen content was influenced by fraction of viable eggs, total clutch sizes, sand temperatures, maximum nest temperature and maximum change in the nest temperature during incubation. Clutch size during embryogenesis was the most influential factor overall. However, the major influential factors were different for different developmental stages. In the first half of the incubation, the development rate was low, and the change in the nest oxygen content was influenced mainly by the clutch size. During the second half, the rapid embryonic development rate became the dominant factor, and hatchling activities caused even greater oxygen consumption during the last stage of development.