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Microbiological characterization of gut symbiotic bacteria in a limited number of stinkbugs of the families Acanthosomatidae, Plataspidae, Pentatomidae, Scutelleridae, Parastrachiidae, Alydidae and Pyrrhocoridae has shown symbiotic association with midgut bacteria to be common in phytophagous taxa of these heteropteran insects. Here we investigated the midgut bacterial symbiont of Eucorysses grandis, a stinkbug of the family Scutelleridae. A specific gammaproteobacterium was consistently identified in insects from five different geographic origins. The bacterium was detected in 64 of 64 insects sampled from three host populations. Phylogenetic analyses revealed that the bacterium constitutes a distinct lineage in the Gammaproteobacteria, neither closely related to the gut symbiont of another scutellerid stinkbug, Cantao ocellatus, nor to gut symbionts of other stinkbugs. Diagnostic PCR, in situ hybridization and electron microscopy demonstrated that the bacterium is located extracelluarly, in the midgut fourth section, which possesses crypts. These results indicate that the primary gut symbionts have multiple evolutionary origins in the Scutelleridae. A Sodalis-allied facultative symbiont was also identified in some insects from natural populations. Biological aspects of the primary gut symbiont and the secondary Sodalis-allied symbiont are discussed.
The age structure of breeding females of Hynobius nebulosus has not been studied sufficiently. We estimated the ages of 76 individuals from a population in Kyoto by using skeletochronology. The mean age and snout-vent length (SVL) of this population were 4.6 years and 55.7 mm, respectively. It was estimated that the youngest females breed two years post hatching at a mean SVL of 46.5 mm, but a larger number of individuals begins breeding at three years and a mean SVL of 52.2 mm. Because most males also start to breed at three years, there seems to be no gender difference in the timing of sexual maturation. The age of the oldest female was estimated to be 11.8 years. It is possible that the life history of H. nebulosus is characterized by early maturation and arrested growth, and short longevity.
The profiles of sex steroids (estradiol-17β, testosterone and 11-ketotestosterone) and the mRNA levels of gonadotropins (luteinizing hormone and follicle-stimulating hormone) were investigated before and after downstream migration in females of the Japanese eel species Anguilla japonica, which were collected in the brackish Hamana Lake and its inlet freshwater rivers. Eels were separated into three groups using otolith microchemistry: ‘migrants’ that grew in the inlet rivers and then made a downstream migration to Hamana Lake mainly in October and November; ‘non-migrant’ yellow eels caught in rivers during the same season; and ‘residents,’ which were yellow eels caught in rivers in August. Sex steroid levels, especially those of testosterone and 11-ketotestosterone, were higher in migrants than in non-migrants and residents. Real-time quantitative PCR analysis indicated that mRNA levels of luteinizing hormone (LH) β-subunits were significantly higher in migrants than in other groups, whereas those of follicle-stimulating hormone β-subunits did not show significant changes during downstream migration. The high levels of these hormones during downstream migration raise the question about if they also play a role in motivating the migratory behavior of eels.
Heparan sulfate (HS), a polysaccharide ubiquitously expressed in animals, is essential for development and homeostasis. Degradation of HS by heparanase, an endoglucuronidase, may affect pathophysiological function. Expression of the heparanase gene has been found elevated in a number of pathological conditions. The goal of this work was to investigate the impact of heparanase on expression of other genes. DNA microarray analysis revealed that 1, 042 genes in the cortex and 1,039 genes in the thalamus are up- or down-regulated more than 2-fold in mouse brain overexpresssing human heparanase. Of these genes, two of the early growth response genes, Egr1 and Egr2, are substantially upregulated in the cortex, but essentially unchanged in the thalamus. RT-PCR analysis demonstrated a significant increase of Egr2, but a minor increase of Egr1, in human embryonic kidney cells stably overexpressing heparanase. The upregulated expression of Egr genes is also observed in hepatoma cells with upregulated expression of heparanase. Earlier studies reported that Egr1 induced heparanase expression; our findings suggest a possible reciprocal regulation of Egr and heparanase expression. Furthermore, overexpression of heparanase influenced expression of most genes involved in heparan sulfate proteoglycan biosynthesis, albeit to a different degree in the cortex and thalamus of the transgenic mice.
Locomotor activity rhythms of dark stock flies of Drosophila melanogaster kept in complete darkness for 700 to 1340 generations were examined. The stock was established by the late Prof. S. Mori in November 1954 to investigate long-term effects of darkness on organisms. The activity of flies was recorded under three types of light conditions: DD after LD12:12, and DD after exposure to a 3.5 h (P3) or 7.5 h (P7) light pulse. In all of these conditions, the experimental dark flies exhibited clear circadian rhythms similar to those of control light flies. We compare our results with those of various studies on troglobites.
Terpios hoshinota is a black-colored sponge encrusting both live and dead corals. On examination, we determined that in the mesohyl, unicellular cyanobacteria were distributed extracellularly in dense populations; roughly half of the area in the histological sections was occupied by the cyanobacterial cells in the mesohyl of the inner zone. Pigment extraction tests of whole sponges and microspectrophotometrical analysis of cyanobacterial symbionts showed that the black coloration of the sponge is attributable to extremely densely packed cyanobacteria expressing R-phycoerythrin. Spermatic follicles were found in the mesohyl of specimens collected in summer, indicating seasonal reproduction, which is likely to play a crucial role in long-distance dispersal. We also found a possible oocyte that did not contain cyanobacteria in the ooplasm.
Bioconvection emerges in a dense suspension of swimming protists as a consequence of their negative-gravitactic upward migration and later settling as a blob of density greater than that of water. Thus, gravity is an important parameter governing bioconvective pattern formation. However, inconsistencies are found in previous studies dealing with the response of bioconvection patterns to increased gravity acceleration (hypergravity); the wave number of the patterns has been reported to decrease during the hypergravity phases of parabolic aircraft flight, while it increases in centrifugal hypergravity. In this paper, we reassess the responses of bioconvection to altered gravity during parabolic flight on the basis of vertical and horizontal observations of the patterns formed by Tetrahymena thermophila and Chlamydomonas reinhardtii. Spatiotemporal analyses of the horizontal patterns revealed an increase in the pattern wave number in both pre- and post-parabola hypergravity. Vertical pattern analysis was generally in line with the horizontal pattern analysis, and further revealed that hypergravity-induced changes preceded at the top layer of the suspensions while microgravity-induced changes appeared to occur from the bottom part of the settling blobs. The responses to altered gravity were rather different between the two sample species: T. thermophila tended to drastically modify its bioconvection patterns in response to changes in gravity level, while the patterns of C. reinhardtii responded to a much lesser extent. This difference can be attributed to the distinct physical and physiological properties of the individual organisms, suggesting a significant contribution of the gyrotactic property to the swimming behavior of some protists.
Alpha-fetoprotein (AFP) is a fetal glycoprotein that is known as a biomarker for monitoring pregnancy in many mammalian species. However, characterization of AFP has not yet been undertaken in any cetacean species. Here, we purified AFP from the serum of fetal striped dolphin by chemical precipitation followed by a combination of immunoadsorbent column chromatography and gel filtration. The molecular masses of native and denatured dolphin AFP were estimated to be ∼78,000 Da by gel filtration and ∼68,000 Da by SDS-PAGE, respectively, representing typical masses reported for mammalian AFPs. In fetal serum, only the AFP band (∼68,000 Da) appeared to be immunoreactive to an antiserum against purified dolphin AFP, indicating sufficient specificity for the development of an AFP immunoassay. Full-length cDNA encoding for the dolphin AFP was cloned from fetal liver and revealed an open reading frame comprising 610 amino acid residues, which included a putative signal peptide of 18 amino acid residues. This was followed by a sequence identical to the N-terminus of purified AFP. The deduced amino acid sequence of dolphin AFP showed more than 80% identity to those of other mammalian AFPs. To our knowledge, the present report represents the first identification and characterization of AFP from any cetacean species.
The burrows created by benthos in tidal flats provide various habitats to other organisms. Echiuran burrows are unique among these in being persistently disturbed by the host's undulating activity, but little is known on how symbionts adapt to such a unique habitat. We report here the morphological and ecological adaptation by two bivalve species of Basterotia (Sportellidae), including one new species, which are commensals with burrowing echiuran worms. The burrows of Ikedosoma gogoshimense were inhabited by Basterotia gouldi at intertidal gravelly mud flats in the central Seto Inland Sea, whereas those of Ochetostoma erythrogrammon were inhabited by Basterotia carinata n. sp. at an intertidal gravelly coral-sand flat at Amami-Ohshima Island. Both bivalve species were found embedded in the burrow wall with their posterior inhalant and exhalant apertures gaping to the burrow lumen, suggesting that they utilize the water currents created by host echiurans. The posteriorly robust, laterally inflated shell with developed carina is considered an adaptation to symbiotic life, as it is exposed to pressure caused by the host's persistent undulating activity. Females of Basterotia bivalves were larger than males, suggesting size-dependent sex change, and possessed brooded veligers in the ctenidium. Our findings suggest that species-specific intimate association with echiurans may be widespread among the Sportellidae bivalves, whose biology remains poorly understood.
A new species of the genus Rhinolophus is described from Yunnan Province, southwestern China. The new taxon belongs to the Rhinolophus “philippinensis-group” and is distinguished by differences in the nose-leaf structures, craniodental characteristics, and bacular features.
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