We examined the compensational recovery of the response rate (relative occurrence) of the wind-evoked escape behavior in unilaterally cercus-ablated crickets (Gryllus bimaculatus) and elucidated the existence of a sensitive period for such recovery by rearing the crickets under different conditions. In one experiment, each cricket was reared in an apparatus called a walking inducer (WI) to increase the sensory input to the remaining cercus, i.e., the self-generated wind caused by walking. In another experiment, each cricket was reared in a small plastic case separate from the outside atmosphere (wind-free: WF). In this rearing condition, the cricket did not experience self-generated wind as walking was prohibited. During the recovery period after the unilateral cercus ablation, the crickets were reared under either the WI or WF condition to investigate the role of the sensory inputs on the compensational recovery of the response rate. The compensational recovery of the response rate occurred only in the crickets reared under the WI condition during the early period after the ablation. In particular, WI rearing during the first three days after the ablation resulted in the largest compensational recovery in the response rate. In contrast, no compensational recovery was observed in the crickets reared under the WF condition during the first three days. These results suggest that a sensitive period exists in which sensory inputs from the remaining cercus affect the compensational recovery of the response rate more effectively than during other periods.

To elucidate the mechanism for preventing entry into embryonic diapause or breakdown of diapause in Bombyx mori by HCl and dimethyl sulfoxide (DMSO) treatment or a combination of cold and HCl treatment, we performed quantitative analysis of Ca² and Mg² in the chorion and egg content using inductively coupled plasma atomic emission spectrometry (ICP-AES). When diapause eggs that had been incubated at 25°C for 2 days from oviposition and at 4°C for an additional six days were treated with HCl solution, the amount of Ca² in the chorion and egg content after HCl treatment was reduced to one-seventh, as compared with the amount before treatment. In contrast, there was no change in the amount of Mg² with HCl treatment. The amount of Ca² in the HCl solution after the diapause eggs were treated increased 7.5-fold, as compared with that of eggs treated with water. Even when 17-day-old diapausing eggs were treated with HCl, which did not break diapause, the amount of Ca² in the chorion and egg content was reduced to one-fifth, as compared with the control. Meanwhile, changes in Ca² and Mg² contents were not observed in 12-hr-old diapause-destined eggs before or after treatment with DMSO, which effectively prevents diapause. These data may suggest that Ca² efflux from diapause eggs by HCl is not directly associated with preventing entry into diapause or breaking of diapause. In addition, we discovered that the amount of Ca² in diapause-destined eggs was more than 2.4-fold larger than in non-diapause-destined eggs.

A previous study by our group reported that mouse and human myoblasts fail to express myogenin and to fuse into multi-nucleate myotubes when cultured at low temperature, such as 30°C, but that this activity is rescued by adding IGF-I and vitamin C to the culture medium. In the present study, we examined mitochondrial activity as a target of the inhibitory effects of the low culture temperature. It has been suggested that mitochondria regulate myogenesis. By using a mouse myoblast cell line C2C12, we demonstrate that the expression of cytochrome c oxidase subunit I (COX I), which is encoded in mitochondrial genome, increases during myogenic differentiation at the normal culture temperature (38°C), but that this up-regulation is inhibited at 30°C. The mitochondrial membrane potential also decreased at 30°C compared to the culture at 38°C. However, IGF-I and vitamin C rescued both COX I expression and mitochondrial membrane potential at 30°C as promoting muscle differentiation. We also find that the rescue of mitochondrial activity by IGF-I and vitamin C at 30°C occurred after the myogenin expression, which suggests that myogenin regulates mitochondrial function during myogenesis. We suggest that our low temperature-culture system may be suitable for use in studying the detailed mechanism of myogenin-related phenomena during myogenesis.

Eggshell evaluation may serve as an indicator of the effect of substances released in the environment, which may change eggshell shape, size, structure, and/or chemical composition. Additionally, exposure may interfere with hatching rates in contaminated eggs. The objective of this study was to better understand how exposure to the insecticide methyl parathion interferes with chemical changes in eggshells of Podocnemis expansa throughout their artificial incubation, as well as with egg hatchability. A total of 343 P. expansa eggs were collected in a natural reproduction area for the species. These eggs were transferred to and artificially incubated in the Wild Animal Teaching and Research Laboratory at Universidade Federal de Uberlândia. On the first day of artificial incubation, 0, 35, 350, and 3500 ppb of methyl parathion were incorporated to the substrate. Eggs were collected every three days for chemical analysis of eggshells. Hatchability was evaluated as the number of hatchlings in each treatment, for the eggs that were not used in the chemical analysis. Student's T-test was used for data on eggshell chemical composition, and the Binomial Test for Two Proportions was used in the hatchability analysis, at a 5% significance level. It was observed that the incorporation of methyl parathion to the substrate on the first day of artificial incubation of P. expansa eggs reduced the levels of total fat in the shells throughout their incubation, besides reducing egg hatchability.

Drosoph1la ornatifrons of the guarani group (Diptera: Drosophilidae) is found mainly in humid areas of the Atlantic Forest biome, especially in the southern region of Brazil. Historical and contemporary fragmentation events influenced species diversity and distribution in this biome, although the role of paleoclimatic and paleogeographic events remain to be verified. The objective of the present study was to evaluate the demographic structure of D. ornatifrons from collection sites that are remnants of Atlantic Forest in southern Brazil, in order to contribute to the understanding of the processes that affected the patterns of genetic variability in this species. To achieve this goal, we sequenced 51 individuals from nine localities and 64 individuals from six localities for the mitochondrial genes Cytochrome Oxidase I and II, respectively. Our results indicate that D. ornatifrons may have experienced a demographic expansion event from the southernmost locations of its distribution, most likely from those located next to the coast and in fragments of Atlantic Forest inserted in the Pampa biome (South 2 group), towards the interior (South 1 group). This expansion probably started after the last glacial maximum, between 20,000 and 18,000 years ago, and was intensified near the Pleistocene-Holocene transition, around 12,000 years ago, when temperature started to rise. In this work we discuss how the haplotypes found barriers to gene flow and dispersal, influenced by the biogeographic pattern of Atlantic Forest.

Eremias multiocellata is a viviparous lizard that is known to exhibit temperature-dependent sex determination (TSD). Conventional Giemsa staining under light microscope examination has identified the karyotype of this species to be 2 n = 36 I 2 m, with no detectable heteromorphic sex chromosomes. However, a highly differentiated female-specific chromosome, W, which is homomorphic with the Z chromosome, is found in the present study by the high-resolution cytogenetic method of comparative genomic hybridization (CGH). The results show that E. multiocellata is a viviparous lizard with both TSD and ZW heterogametic sex chromosomes. Despite the fact that a different sex ratio of male offspring was found in two populations (separated by an altitude of 1400 m) in previous incubation experiments, we demonstrate, using genomic in situ hybridization (GISH), that there is no significant chromosomal loss or acquisition between the two populations. This suggests that temperature may play a more important role in lowland populations. These results most likely indicate that E. multiocellata is transitioning between the evolutionary processes of TSD and genotypic sex determination (GSD) systems, and also give clues to the effect of TSD versus GSD in this process.

Medium-sized carnivorous mammals are important seed dispersers of fleshy fruits. The raccoon dog (Nyctereutes procyonoides viverrinus) often feeds on fleshy fruits and forms latrines. This behavior may potentially lead to seed dispersal. To determine if this is the case, we studied 1) seed recovery in the droppings of raccoon dogs, and 2) the transportation of seeds between habitats using plastic markers in a western suburb of Tokyo, Japan. In total, 32,473 seeds of 50 plant taxa were recovered from 120 raccoon dog droppings during a year, and 95.7% of the seeds were found to be those of fleshy fruits. The species most frequently recovered were the eurya (Eurya japonica, 52.6%), the brambles (Rubus spp., 17.4%), and the black night shade (Solanum nigrum, 16.0%). A total of 7,412 plastic markers were embedded in baits at 14 bait plots and were recovered in the feces of the raccoon dogs at 22 latrines. The “transportation rates” were calculated in 50-m distance classes and found that most seeds (43.5%) were deposited within 50 m from the bait point, suggesting very short seed dispersal distances. Inter-habitat transportation was observed: 64.9% of the retrieved markers deposited in the forest were transported to other places within the forest. In contrast, almost all of the markers (99.4%) deposited in the open site were transported within the same habitat. These findings suggest that the seeds of forest plants bearing berries can be dispersed out of the forest to open areas by raccoon dogs.

The aim of this study was to analyze the histology and the histochemical distribution of glycoproteins (GPs) and lipids of the hepatopancreas of Cyrtograpsus angulatus and Neohelice granulata acclimated to salinities of 10 psu (hyperregulation) and 35 psu (osmoconformation). Sections of the hepatopancreas of adult male crabs were treated with hematoxylin-eosin; Masson trichrome; Alcian Blue (pHs 2.8, 1.0, 0.5); Toluidine Blue (pHs 5.6, 4.2); periodic acid Schiff; Sudan Black and Red. At salinity 35 psu, the hepatopancreas of both species exhibited typical histological features, whereas at salinity 10 psu, detachment of the basal lamina, desquamated epithelium, disrupted brush border, loss of intercellular cohesion, hypertrophied tubular lumen, and hemolymph infiltration between cells were observed in some zones. Resorptive cells (R-cells) and vacuoles of blister-like cells (B-cells) of both species show a higher glycogen content at 35 psu than at 10 psu. At lower salinities, the cytoplasm of the different cell types evidence higher contents of carboxylated GPs in N. granulata and of sulfated GPs in C. angulatus. At both salinities, and at the two pHs in N. granulata and at pH 5.6 in C. angulatus, the brush border, the vacuoles of B-cells and the peritrophic membrane show metachromasia. R-cell vacuoles and the cytoplasm of all cell types —except for the E-cells—at all salinities in both species show abundant lipid droplets. The results of the present study contribute significant data to the histophysiology of crustacean decapods, favoring the comprehension of the complex adjustment mechanisms facing saline stress in euryhaline crabs.

Feeding adaptations are a conspicuous feature of avian evolution. Bill and cranial shape as well as the jaw muscles are closely related to diet choice and feeding behaviors. Diurnal raptors of Falconiformes exhibit a wide range of foraging behaviors and prey preferences, and are assigned to seven dietary groups in this study. Skulls of 156 species are compared from the dorsal, lateral and ventral views, by using geometric morphometric techniques with those landmarks capturing as much information as possible on the overall shape of cranium, bill, orbits, nostrils and attachment area for different jaw muscles. The morphometric data showed that the skull shape of scavengers differ significantly from other raptors, primarily because of different feeding adaptations. As a result of convergent evolution, different scavengers share generalized common morphology, possessing relatively slender and lower skulls, longer bills, smaller and more sideward orbits, and more caudally positioned quadrates. Significant phylogenetic signals suggested that phylogeny also played important role in shape variation within scavengers. New World vultures can be distinguished by their large nostrils, narrow crania and small orbits; Caracaras typically show large palatines, crania and orbits, as well as short, deep and sharp bill.

To analyze the phylogenetic relationships of the genus Pseudocrangonyx within the superfamily Crangonyctoidea, we sequenced a partial LSU rDNA gene from four species (six specimens). The monophyly of Pseudocrangonyx and its affinity to Crymostygius were strongly supported by four reconstruction methods: neighbor-joining, minimum evolution, maximum-likelihood, and Bayesian inference. Current geographical distributions of the taxa studied and their phylogenetic relationships established in our study suggest that members of Pseudocrangonyctidae and Crymostygius diverged from a common Crangonyx-like ancestor in the Arctic Basin region of the northern hemisphere.

Social environments often affect the development of organisms. In Tenebrionidae beetles, larval development can be arrested at the final instar stage in the presence of conspecific larvae. This developmental plasticity is considered to be an anti-cannibalistic strategy but the critical environmental determinants and actual effects remain to be elucidated. In this study, we examined the effects of the heterospecific environment, conspecific sexual environment (i.e., presence of conspecific male or female), and abiotic physical stimulation on the pupation decision of the sexually dimorphic horned-flour beetle Gnatocerus cornutus. Additionally, actual anti-cannibalistic or antipredatory effects of developmental arrest were evaluated by analyzing stage-dependent vulnerabilities. When G. cornutus larvae were maintained with a G. cornutus larva, a G. cornutus adult, or T. castaneum adult, the developmental period up to the prepupal stage was significantly elongated, suggesting that the cue is not species-specific. Sexual environment did not affect the timing of pupation in G. cornutus; however, we found that abiotic tactile stimulations by glass beads could repress pupation. We also discovered that prepupal and pupal stages were more vulnerable to cannibalism and predation than the larval stage. These data suggest that G. cornutus larvae use non-species specific tactile stimulation as a decision cue for pupation and it has broader defensive effects against heterospecific predation as well as conspecific cannibalism.

This paper describes the characteristics of male and female germ cells during gonadal development and the gonadal maturity scale of the blue crab, Callinectes sapidus. A total of 20 specimens were collected monthly from June to November 2012, in two areas off the coast of the Gulf of Mexico: the San Andrés Lagoon and Alvarado Lagoon. The gonads were removed and processed following the standard technique of hematoxylin and eosin staining. An important event in oogenesis (pre-vitellogenesis) was the appearance of a perinuclear vesicle in the cytoplasm and the accumulation of yolk granules. Later, vitellogenesis began and there was an accumulation of nutritive droplets and fragmentation of the perinuclear vesicle. During spermatogenesis, the accumulation of two fluids was observed that were involved in the formation of the spermatophore and the delay of spermiogenesis. Based on the histological features of gonad maturity, five stages were described (inactive, early gametogenesis, development, maturity, and resorption), in females and males. This proposal can be useful for the study of reproductive seasonality of this species.

Paramacrochiron chironecicola n. sp. (Copepoda: Cyclopoida: Macrochironidae) is described from the highly venomous box jellyfish Chironex sp. collected from Malampaya Sound, Palawan Island, The Philippines. This is the first record of copepods associated with cubozoan medusae, although other cnidarian groups such scyphozoans, hydrozoans, and anthozoans are common hosts for symbiotic copepods. The infection sites were on the subumbrella, pedalium, and rhopalium, but also rarely on the adradial furrow. The new species is distinguished from other congeners by a combination of the following features: (1) the fifth pedigerous somite dorsally covering the anterior part of the female genital double-somite; (2) the fine structures of the antenna (relative lengths of segments) and maxilliped (positions of terminal elements) of the female; (3) the relatively long outer spines on the exopodal segments of legs 1–4; (4) the relatively long and thick female leg 5 bearing a long protopodal seta which reaches to the distal margin of the exopod; (5) the relatively short caudal ramus in the female; and (6) the plump prosome and short urosome in the male. Since members of the genus typically parasitize scyphozoans, especially rhizostomes, the association of this parasitic copepod on cubozoans may reflect the relatively close phylogenetic relationship between cubozoans and scyphozoans.

We report a new tree frog of the genus Gracixalus from western Thailand and describe it as a new species Gracixalus seesom based on results of morphological and molecular analyses. The new species is a small-sized Gracixalus (male snout-vent length ca. 22 mm) and is morphologically similar to G. gracilipes, but is easily distinguished from it by its dorsal tan color in life, absence of white spot on lower lip, and black markings on its foot webbing. The new species also clearly differs from all the other members of the genus by the combination of small body size, triangular snout, and light yellowish brown dorsum without distinct tuberculations. Problems of phylogeny and taxonomy of the genus Gracixalus are briefly discussed.

The Crinoidea are the most primitive class of living echinoderms, and suffered a severe crisis during the Late Permian mass extinction event. All post-Palaeozoic crinoids, including living species, belong to the Articulata, and morphological and recent molecular studies demonstrate that they form a monophyletic clade. The Articulata originated from Palaeozoic cladid crinoids, but the nature and timing of their origination remains obscure. Problems with understanding the origin and early evolution of the Articulata have arisen because the Permian—Triassic crinoid fossil record is particularly poor. We report on a new genus and species from the earliest Triassic, which is the oldest known post-Palaeozoic articulate crinoid and fundamentally alters our understanding of the early evolution of the Articulata. Prior to this study, the most primitive post-Palaeozoic articulate was thought to be Holocrinus of the order Isocrinida. Unexpectedly, the new taxon belongs to the order Encrinida, which reveals a previously hidden diversity of crinoids in the earliest Triassic. Its discovery implies either a dramatic radiation of crinoids in the immediate post-extinction aftermath, when environmental conditions were at their most severe, or a pre-extinction origin of the crown group articulates and survival of multiple lineages.