Volumes & issues:
33, 4(2016); http://dx.doi.org/10.2108/zs150077View Description Hide Description
This study describes the reproductive microhabitat of Melanophryniscus montevidensis and its use in two water bodies (WBs) in Barra de la Laguna de Rocha, Uruguay. Monthly field trips were performed between March 2012 and February 2013. Variables related to the WBs and vegetation, as well as parameters linked to the usage the amphibians make of the site (e.g: distance to the border of the pond, water depth and the vegetation use) were recorded. The behavior shown by the individuals during the breeding activity was recorded. This activity occurs in shallow temporary WBs with abundant hydrophilic vegetation. The individuals were found more frequently in areas near the edge of the pond, which has denser vegetation. The calling males were found closer to the border of the pond, and they showed better body condition than the non-calling males. In addition to calling activities, males used alternative tactics to find couples, such as active search of females, and aggressive behaviors, such as male displacing and physical combat. Such behaviors are common in anurans with explosive reproductive dynamics. The characterization of the reproductive microhabitats permits the proposal of strategies for the conservation of the species in Uruguay, given that the loss and fragmentation of habitats is one of the main causes considered for the decrease in their populations.
The Size of Winter-Flooded Paddy Fields No Longer Limits the Foraging Habitat Use of the Endangered Crested Ibis (Nipponia nippon) in Winter33, 4(2016); http://dx.doi.org/10.2108/zs150012View Description Hide Description
Paddy fields have traditionally been viewed as the key foraging habitats for the endangered crested ibis (Nipponia nippon). With the population of this species now increasing, its distribution has expanded to both lowland areas and outside the nature reserve. However, little is known about the current foraging habitat preferences of these birds, especially during winter. In this research, a total of 54 used sites and 50 unused sites were investigated during winter from December 2011 to January 2012. The results of logistic regression analysis indicate that soil softness, human disturbance, and distance to the nearest road were important factors. For the site plots of winter-flooded paddy fields, the birds prefer the paddy fields with higher coverage of vegetation, except softer foraging sites and lower human-related disturbance. In lowland areas, the size of winter-flooded paddy fields was not a limiting factor, due to the availability of other wetlands capable of providing abundant food. The micro-habitat characteristics were important indicators of foraging habitat quality rather than the size of winter-flooded paddy fields, and the food accessibility may play an important role in the process of foraging habitat use. We suggest the improvement of the foraging micro-habitat and environmental characteristics would be effective in ensuring the availability of food in the dispersed lowland areas. The local people still needed to be encouraged and compensated by their single-cropping cultivation, ploughed the paddy fields after harvesting and irrigated them with shallow water flooded in the original core areas of the nature reserve.
Estimated Seed Shadow Generated by Japanese Martens(Martes melampus): Comparison with Forest-Dwelling Animals in Japan33, 4(2016); http://dx.doi.org/10.2108/zs160017View Description Hide Description
To evaluate the seed shadow generated by wild Japanese martens (Martes melampus), we combined data on their ranging behavior from the northern foot of Mt. Fuji, central Japan (seven males and three females) with data on gut passage time obtained from martens in Toyama Municipal Family Park Zoo (three males and one female). The movement distances varied, and mean distances for 0–1, 2–3, and 4–5 h intervals were 152.4, 734.7, and 1,162.4 m, respectively, with no significant sex difference. The mean gut passage time of ingested seeds was 7.4 h (range: 0.6–51.7 h), and two-thirds were defecated within 12 h. Seeds of fleshy fruits was frequently transported to 501–1,000 m, and 20% of ingested seeds were transported > 1,000 m from feeding sites. We found positive correlations between body size and home range of the animals in Japan and their seed dispersal distances. We conclude that Japanese martens are medium-range dispersers that can transport seeds from the source to open habitats conducive for germination and/or growth, partly due to scent marking behaviors.
Comparative Analysis of Genome and Epigenome in Closely Related Medaka Species Identifies Conserved Sequence Preferences for DNA Hypomethylated Domains33, 4(2016); http://dx.doi.org/10.2108/zs160030View Description Hide Description
The genomes of vertebrates are globally methylated, but a small portion of genomic regions are known to be hypomethylated. Although hypomethylated domains (HMDs) have been implicated in transcriptional regulation in various ways, how a HMD is determined in a particular genomic region remains elusive. To search for DNA motifs essential for the formation of HMDs, we performed the genome-wide comparative analysis of genome and DNA methylation patterns of the two medaka inbred lines, Hd-rRII1 and HNI-II, which are derived from northern and southern subpopulations of Japan and exhibit high levels of genetic variations (SNP, ∼ 3%). We successfully mapped > 70% of HMDs in both genomes and found that the majority of those mapped HMDs are conserved between the two lines (common HMDs). Unexpectedly, the average genetic variations are similar in the common HMD and other genome regions. However, we identified short well-conserved motifs that are specifically enriched in HMDs, suggesting that they may play roles in the establishment of HMDs in the medaka genome.
- Molecular Biology
33, 4(2016); http://dx.doi.org/10.2108/zs150186View Description Hide Description
In most animals, aging is an irreversible process; however the species Turritopsis sp. has been observed to undergo a rejuvenation process as many as 14 times. In the present study, we used multiplexed RNA libraries to obtain the transcriptome from four developmental stages (St) of Turritopsis sp., including (I) immature medusa, (II) dumpling, (III) dumpling with a short stolon, and (IV) polyp, which had recently rejuvenated. A total of 4.02 billion paired-end reads were assembled de novo, yielding 90,327 contigs. Our analyses revealed that significant blast hits were recovered for 74% of the assembled contigs, and 19% were successfully annotated with gene ontology (GO) terms. A BLAST search demonstrated that 32% of the contigs were most similar to Hydra vulgarissequences. Raw reads from each sample were mapped against the contigs to find St-specific genes. This represents the first comprehensive set of de novo transcriptome data for this species, which may provide clues toward a better understanding of cyclical rejuvenation in multicellular animals.
33, 4(2016); http://dx.doi.org/10.2108/zs150130View Description Hide Description
The objective of the present study was to analyze chondrogenesis and the ossification pattern of the limbs of Melanosuchus niger in order to contribute with possible discussions on homology and the fusion pattern of autopodial elements and phylogeny. In the Reserva Extrativista do Lago Cuniã, Rondônia, Brazil, six nests were marked and two eggs removed from each nest at 24-hour intervals until hatching. Embryos were cleared using KOH; bone tissue was stained with alizarin red S and cartilage with Alcian blue. Routine staining with HE was also performed. In the pectoral girdle, the scapula showed ossification centers before the coracoid process. In the pelvic girdle, the ilium and the ischium were condensed as a single cartilage, although ossification took place through two separate centers, forming distinct elements in the adult. The pubis developed from an independent cartilaginous center with free end, which reflects its function in breathing. In the initial stages, the stylopodium and the zeugopodium developed from the condensation of a Y-shaped cartilage in the limbs, and differentiation of the primary axis and digital arch were observed. The greatest changes were observed in the mesopodia. In their evolution, Crocodylia underwent a vast reduction in the number of autopodial elements as a consequence of fusions and ossification of some elements. This study shows that the chondrogenesis and ossification sequences are dissociated. Moreover, the differences between M. niger and other species show clear variation in the patterns for these events in Alligatoridae.
Comparative Morphological Analysis of the Immature Stages of the Grass Blue Butterflies Zizeeria and Zizina (Lepidoptera: Lycaenidae)33, 4(2016); http://dx.doi.org/10.2108/zs150171View Description Hide Description
The pale grass blue butterfly has been used to assess the biological effects of the Fukushima nuclear accident. Zizeeria and Zizina are two closely related genera of grass blue butterflies that are widely distributed in tropical to temperate Asia, Australia, and Africa, making them suitable environmental indicators for these areas. However, the morphological features of the immature stages have been examined only in fragmentary fashion. Here, we reared Zizeeria maha argia, Zizeeria maha okinawana, Zizeeria karsandra karsandra, Zizina emelina emelina, Zizina otis labradus, and Zizina otis riukuensis using a standard rearing method that was developed for Zizeeria maha, and comparatively identified morphological traits to effectively classify the immature stages of species or subspecies. Morphological information on these and other subspecies including Zizeeria knysna knysna and Zizina otis antanossa from Africa was also collected from literature. The subspecies were all reared successfully. The subspecies all had dorsal nectary and tentacle organs with similar morphology. For the subspecies of Zizeeria maha, only minor morphological differences were noted. Similarly, the subspecies of Zizina otis shared many traits. Most importantly, Zizeeria and Zizina differed in the shape of the sensory hairs that accompany the dorsal nectary organ; Zizeeriahad pointed hairs, and Zizina had blunt or rounded hairs. However, Zizina emelina exhibited several intermediate features between these two genera. Overall, the morphological traits did not completely reflect the conventional systematic relationships. This comparative study describes the efficient rearing of the grass blue butterflies and provides a morphological basis for the use of these species as environmental indicators.
Phylogenetic Relationships of Japanese Auritibicen Species (Hemiptera: Cicadidae: Cryptotympanini) Inferred from Mitochondrial and Nuclear Gene Sequences33, 4(2016); http://dx.doi.org/10.2108/zs160007View Description Hide Description
We investigated the phylogenetic relationships and divergence times within the genus Auritibicen(Cicadidae: Cicadinae: Cryptotympanini), analyzing five Japanese species (A. japonicus, A. bihamatus,A. kyushyuensis, A. esakii and A. flammatus) and three species from East Asian mainland and Taiwan (A. atrofasciatus, A. intermedius and A. chujoi) using mitochondrial cytochrome oxidase subunit I (COI) and nuclear elongation factor 1-alpha (EF-1a) gene sequences. Although the EF-1a gene tree did not resolve the relationships among these Auritibicen species, the trees based on COI gene and the combined data set showed that Japanese taxa comprised three distinct lineages: the individual species A. flammatus and A. bihamatus, and the A. japonicus group, comprising A. japonicus, A. esakii and A. kyushyuensis from Japan and A. intermedius from Korea. In A. kyushyuensis, which comprises three populations in Kyushu, western Honshu and Shikoku, the specimens from western Honshu and Shikoku were closely related to each other, but not to the specimen from Kyushu; instead, they were sister to the Korean A. intermedius. The incongruence between the gene tree and species tree necessitates further population genetic and morphological studies to confirm the classification and species status of the western Honshu and Shikoku populations of A. kyushyuensis, which were originally described as two independent species. Divergence time estimation suggested that the most recent common ancestor of Auritibicen species studied dated back to the late Pliocene and that the species of the A. japonicus group diverged during the mid Pleistocene. Thus, the Pleistocene climatic fluctuation may have promoted the divergence of the Auritibicen species.
Seawater Polluted with Highly Concentrated Polycyclic Aromatic Hydrocarbons Suppresses Osteoblastic Activity in the Scales of Goldfish, Carassius auratus33, 4(2016); http://dx.doi.org/10.2108/zs150211View Description Hide Description
We have developed an original in vitro bioassay using teleost scale, that has osteoclasts, osteoblasts, and bone matrix as each marker: alkaline phosphatase (ALP) for osteoblasts and tartrate-resistant acid phosphatase (TRAP) for osteoclasts. Using this scale in vitro bioassay, we examined the effects of seawater polluted with highly concentrated polycyclic aromatic hydrocarbons (PAHs) and nitro-polycyclic aromatic hydrocarbons (NPAHs) on osteoblastic and osteoclastic activities in the present study. Polluted seawater was collected from two sites (the Alexandria site on the Mediterranean Sea and the Suez Canal site on the Red Sea). Total levels of PAHs in the seawater from the Alexandria and Suez Canal sites were 1364.59 and 992.56 ng/l, respectively. We were able to detect NPAHs in both seawater samples. Total levels of NPAHs were detected in the seawater of the Alexandria site (12.749 ng/l) and the Suez Canal site (3.914 ng/l). Each sample of polluted seawater was added to culture medium at dilution rates of 50, 100, and 500, and incubated with the goldfish scales for 6 hrs. Thereafter, ALP and TRAP activities were measured. ALP activity was significantly suppressed by both polluted seawater samples diluted at least 500 times, but TRAP activity did not change. In addition, mRNA expressions of osteoblastic markers (ALP, osteocalcin, and the receptor activator of the NF-κB ligand) decreased significantly, as did the ALP enzyme activity. In fact, ALP activity decreased on treatment with PAHs and NPAHs. We conclude that seawater polluted with highly concentrated PAHs and NPAHs influences bone metabolism in teleosts.
A Stenohaline Medaka, Oryzias woworae, Increases Expression of Gill Na+, K+-ATPase and Na+, K+, 2Cl- Cotransporter 1 to Tolerate Osmotic Stress33, 4(2016); http://dx.doi.org/10.2108/zs150157View Description Hide Description
The present study aimed to evaluate the osmoregulatory mechanism of Daisy's medaka, O. woworae,as well as demonstrate the major factors affecting the hypo-osmoregulatory characteristics of euryhaline and stenohaline medaka. The medaka phylogenetic tree indicates that Daisy's medaka belongs to the celebensis species group. The salinity tolerance of Daisy's medaka was assessed. Our findings revealed that 20‰ (hypertonic) saltwater (SW) was lethal to Daisy's medaka. However, 62.5% of individuals survived 10‰ (isotonic) SW with pre-acclimation to 5‰ SW for one week. This transfer regime, “Experimental (Exp.) 10‰ SW”, was used in the following experiments. After 10‰ SW-transfer, the plasma osmolality of Daisy's medaka significantly increased. The protein abundance and distribution of branchial Na+, K+-ATPase (NKA) and Na+, K+, 2Cl- cotransporter 1 (NKCC1) were also examined after transfer to 10‰ SW for one week. Gill NKA activity increased significantly after transfer to 10‰ SW. Meanwhile, elevation of gill NKA αα-subunit protein-abundance was found in the 10‰ SW-acclimated fish. In gill cross-sections, more and larger NKA-immunoreactive (NKA-IR) cells were observed in the Exp. 10‰ SW medaka. The relative abundance of branchial NKCC1 protein increased significantly after transfer to 10‰ SW. NKCC1 was distributed in the basolateral membrane of NKA-IR cells of the Exp. 10‰ SW group. Furthermore, a higher abundance of NKCC1 protein was found in the gill homogenates of the euryhaline medaka, O. dancena, than in that of the stenohaline medaka, O. woworae.