Gene expression profiles in the testis associated with testis-ova in adult Japanese medaka (Oryziaslatipes) exposed to 17α-ethinylestradiol

The occurrence of oocytes in the testis (testis-ova) of several fish species is often associated with exposure of estrogenic chemicals. However, induction mechanisms of the testis-ova remain to be elucidated. To develop marker genes for detecting testis-ova in the testis, adult male medaka were exposed to nominal concentration of 100 ng L⁻¹ of 17α-ethinylestradiol (EE2) for 3-5 weeks, and 800 ng estradiol benzoate (EB) for 3 weeks (experiment I), and a measured concentration of 20 ng L⁻¹ EE2 for 1-6 weeks (experiment II). Histological analysis was performed for the testis, and microarray analyses were performed for the testis, liver and brain. Microarray analysis in the estrogen-exposed medaka liver showed vitellogenin and choriogenin as estrogen responsive genes. Testis-ova were induced in the testis after 4 weeks of exposure to 100 ng L⁻¹ EE2, 3 weeks of exposure to 800 ng EB, and 6 weeks of exposure to 20 ng L⁻¹ EE2. Microarray analysis of estrogen-exposed testes revealed up-regulation of genes related to zona pellucida (ZP) and the oocytes marker gene, 42Sp50. Using quantitative RT-PCR we confirmed that Zpc5 gene can be used as a marker for the detection of testis-ova in male medaka.     

Styrene dimers and trimers affect reproduction of daphnid (Ceriodaphnia dubia)

The endocrine disruptor activity of styrene in humans and other vertebrates appears to be negligible. However, offspring numbers were reduced in Ceriodaphnia dubia bred in polystyrene cups. Styrene dimers and trimers were found to be eluted from the polystyrene cups by hexane and methanol with gas chromatography-mass spectrometry. Styrene dimers and trimers at concentrations of 0.04-1.7 microg/l affected C. dubia fertility (25% reduction after seven days), suggesting that styrenes have the potential to impair crustacean populations in the aquatic environment.     

Preparation and characteristics of high performance paper containing titanium dioxide photocatalyst supported on inorganic fiber matrix

A novel paper-based material containing titanium dioxide (TiO₂) photocatalyst was successfully prepared by a papermaking technique with the internal addition of inorganic fibers on which TiO₂ particles were supported. Photodegradation performance of acetaldehyde gas, an indoor pollutant, and the durability of the TiO₂-containing papers were investigated under UV irradiation. Ceramic fiber suspension and polydiallyldimethylammonium chloride as a cationic flocculant were mixed, followed by the addition of TiO₂ suspension and anionic polyacrylamide. Subsequently, the inorganic mixture was poured into a pulp suspension, and TiO₂ handsheets then prepared by a papermaking method. The tensile strength of TiO₂-containing paper without a ceramic carrier decreased by more than 30% after 240-h UV irradiation (2 mW/cm²), although the strength of the TiO₂ sheet with ceramic fibers remained reasonably stable. The efficiency of acetaldehyde decomposition by the TiO₂ paper containing an inorganic carrier was nearly equal to that of the carrier-free TiO₂ paper. Scanning electron microscopic observation suggested that most TiO₂ particles were predominantly supported on the inorganic fiber matrix, and were mostly out of contact with organic pulp fibers. The TiO₂ paper with an inorganic carrier demonstrated both excellent photocatalytic performance and durability, which before had been mutually incompatible for organic materials containing TiO₂ photocatalyst. The two-stage mixing procedure for TiO₂ sheet-making is promising for the simple manufacture of high performance paper with photocatalytic ability.     

Juvenile hormone agonists affect the occurrence of male Daphnia

The water flea Daphnia magna reproduces primarily by cyclic parthenogenesis. Environmental stimuli that signal a change to adverse conditions induce the organisms to switch from parthenogenesis to gamogenetic reproduction. During the gamogenetic period, they produce male daphnids and dormant resting eggs, which can survive prolonged periods of environmental adversity. However, little is known about the mechanisms associated with the switch from parthenogenesis to gamogenetic reproduction. We investigated the effects of several juvenoids on sex determination in Daphnia. Females less than 24 h old were exposed to various concentrations of the test substance and were observed for 21 days. It was found that they can trigger the appearance of male daphnids: the percentage of males in the population increases to a level greater than what occurs under ordinary environmental conditions. We found that methylfarnesoate, juvenile hormone III, methoprene, and the phenoxyphenoxy derivatives pyriproxyfen and fenoxycarb (both insecticides) reduced the production of offspring and produced sex ratios dominated by male daphnids. Pyriproxyfen and fenoxycarb showed striking effects at low concentrations. Exposure to either of these chemicals at a concentration of 330 ngl(-1) caused adult females to produce almost all male neonates. Methylfarnesoate, juvenile hormone III, and methoprene showed an effect in inducing male production at higher concentrations (3.7 x 10(3), 3.3 x 10(5), and 1.3 x 10(5) ngl(-1), respectively). Our findings suggest that juvenile hormone agonists, including some insecticides, affect the chemical signaling responsible for inducing the production of male offspring.     

Why is competition more intense if food is supplied more slowly?

We investigated competition for food among two groups of six clone amagos (salmonids), Oncorhynchusmasoumacrostomus, in a laboratory experiment with different rates of food input. We examined the effect of temporal clumping of food resources on the inequality of food sharing between competitors. Monopolization of food by dominants was greater at a low input rate (one food item per 10 s) than at a high input rate (1 food item per 1 s). Aggressive behavior by dominants was more frequent at the low input rate than at the high input rate; its purpose was presumably to interfere with the feeding behavior of subordinates. We assessed the relative importance of three foraging factors (the number of approaches to food items, the chance per approach and the gain per chance) in enhancing inequality in food gain between individuals. Dominants had a disproportionately high chance per approach and gain per chance at the low input rate, but not at the high input rate. The chance of obtaining a food item per approach depended on how many competitors approached simultaneously. The gain per chance depended on the competitive ability of the approaching fish. There was an interaction between these components, such that the number of approaches affected the chance per approach and gain per chance. We evaluated the independent effect of the chance per approach, and showed that it was higher for dominants than for subordinates at the low input rate, but not at the high input rate. This implies that subordinates changed their behavior and became more likely to avoid approaching food at the same time as dominants at the low input rate. Received: 13 August 1996 / Accepted after revision: 30 November 1996     

Effect of void structure of photocatalyst paper on VOC decomposition

TiO₂ powder-containing paper composites, called TiO₂ paper, were prepared by a papermaking technique, and their photocatalytic efficiency was investigated. The TiO₂ paper has a porous structure originating from the layered pulp fiber network, with TiO₂ powders scattered on the fiber matrix. Under UV irradiation, the TiO₂ paper decomposed gaseous acetaldehyde more effectively than powdery TiO₂ and a pulp/TiO₂ mixture not in paper form. Scanning electron microscopy and mercury intrusion analysis revealed that the TiO₂ paper had characteristic unique voids ca. 10 μm in diameter, which might have contributed to the improved photocatalytic performance. TiO₂ paper composites having different void structures were prepared by using beaten pulp fibers with different degrees of freeness and/or ceramic fibers. The photodecomposition efficiency was affected by the void structure of the photocatalyst paper, and the initial degradation rate of acetaldehyde increased with an increase in the total pore volume of TiO₂ paper. The paper voids presumably provided suitable conditions for TiO₂ catalysis, resulting in higher photocatalytic performance by TiO₂ paper than by TiO₂ powder and a pulp/TiO₂ mixture not in paper form.     

Comparative analysis on the genetic population structures of the deep-sea whelks <Emphasis Type="Italic">Buccinum tsubai</Emphasis> and <Emphasis Type="Italic">Neptunea constricta</Emphasis> in the Sea of Japan

The population structures of Buccinum tsubai and Neptunea constricta were compared using partial sequences of the mitochondrial COI gene. B. tsubai included four genetically distinguishable subpopulations (Hokkaido, Yamagata-Toyama, Yamato Bank, and San’in), which is consistent with the results of our previous study using mitochondrial 16SrRNA sequences. The genetic structure of N. constricta was also clear, but the lineage sorting corresponding to the geographical distribution was not as clear as that of B. tsubai. The difference between the genetic structures of these two species might be due to a difference in either their dispersal abilities or the historic distribution patterns in the Sea of Japan. The fossil record and nested clade analyses support the latter alternative. The four subpopulations of B. tsubai are estimated to have diverged 0.42–1.46 million years ago, providing support for the existence of intermediate water of normal salinity and oxygen concentration capable of sustaining B. tsubai between the low-salinity surface water and anoxic deep water during the Pleistocene glacial periods. The genetic diversity of the Yamato Bank subpopulation of B. tsubai was lower than that of the other subpopulations, which is likely attributable to the small size of the Yamato Bank subpopulation.     

Medaka (Oryzias latipes) for use in evaluating developmental effects of endocrine active chemicals with special reference to gonadal intersex (testis-ova).

Japanese medaka (Oryzias latipes) has been widely used for the evaluation of the toxicity of endocrine active chemicals (EACs) and other chemicals as well as for monitoring the adverse effects of effluent discharges in relation to sexual development and function. It is useful for these evaluations for many reasons including the following: 1) it has a short life cycle facilitating studies extending over long phases of development and over multigenerations, 2) it is easy to rear, 3) male and female phenotypes can easily be distinguished on the basis of secondary sex characteristics, and 4) a genetic marker (DMY) is available for identifying the true genotypic sex. Several biomarkers have been found to be useful for identifying the effects of exposure to estrogenic and androgenic chemicals in medaka and they include increased levels of hepatic vitellogenin (VTG) and testis-ova induction in males for exposure to estrogenic chemicals, and decreased levels of hepatic VTG in females and an altered morphology of dorsal and anal fins and formation of papillae for androgenic chemicals. In this paper, we present a critical analysis of the use of medaka as a test species for studies of endocrine disruption and report on the use of sex-related genetic markers and alterations in gonadal development, including the induction of testis-ova formation, for assessing the disruptive effects of EACs. In this paper, we focus on some of the more recent studies and findings.     

The roach (Rutilus rutilus) as a sentinel for assessing endocrine disruption.

Alterations in development and reproduction as a consequence of exposure to endocrine-disrupting chemicals (EDCs) have been demonstrated in many wildlife species. Animals living in, or closely associated with, the aquatic environment are particularly vulnerable to endocrine disruption because thousands of chemicals are actively disposed into rivers, estuaries and seas. Fish have thus been a focus in endocrine disruption studies, and some of the most comprehensive studies on the disruption of sexual development and function are on the roach (Rutilus rutilus). This paper provides a critical analysis of the roach as a sentinel for studies into endocrine disruption. The paper starts by describing what is known on the basic reproductive biology of the roach, information essential for interpreting chemical effect measures on sexual development and function. We then analyze where and how the roach has been applied to improve our understanding of the estrogenic nature of discharges from wastewater treatment works (WWTWs) and describe the phenomenon of feminized male roach in UK rivers. In this paper, the causation of these effects and issues of relative susceptibility and sensitivity of the roach to the effects of estrogenic EDCs are addressed. The paper then describes the ongoing work on the development of genetic and genomic resources for roach and analyses how these are being applied in studies to understand the mechanisms of disruption of sexual development. Finally, the paper addresses the biological significance of sexual disruption and intersex for the individual and discusses the possible implications for wild populations.