Changes in Age-Related Reproductive Tactics in the Female of the Butterfly, Eurema hecabe

The occurrence of mate solicitation by virgin females was investigated in the butterfly Eurema hecabe. Young (1-day-old) virgin females rarely showed mate solicitation to male model, however, old (at least 6-day-old) virgin females frequently showed such flight. The duration of solicitation was significantly longer in older females than in younger ones. The age-related behavioral change occurs with female oogenesis (Hiroki and Kato 1996), and such behavior may thus be a result of female adaptation to maximize their fecundity. Received: 6 November 1997 / Accepted in revised form: 29 June 1998     

Multivariate analyses of the effect of an urban wastewater treatment plant on spatial and temporal variation of water quality and nutrient distribution of a tropical mid-order river

Freshwater resources are increasingly scarce due to human activities, and the understanding of water quality variations at different spatial and temporal scales is necessary for adequate management. Here, we analyze the hypotheses that (1) the presence of a wastewater treatment plant (WWTP) and (2) a polluted tributary that drains downstream from the WWTP change the spatial patterns of physicochemical variables (pH, turbidity, dissolved oxygen, and electrical conductivity) and nutrient concentrations (reactive soluble phosphorus, total phosphorus, nitrogen series, total nitrogen, and total dissolved carbon) along a mid-order river in SE Brazil and that these effects depend on rainfall regime. Six study sites were sampled along almost 4 years to evaluate the impacts of human activities, including sites upstream (1–3) and downstream (5–6) from the WWTP. The impacts were observed presenting an increasing trend from the source (site 1) towards Água Quente stream (site 4, the polluted tributary), with signs of attenuation at site 5 (downstream from both WWTP and site 4) and the river mouth (site 6). Input of nutrients by rural and urban runoff was observed mainly at sites 2 and 3, respectively. At sites 4 and 5, the inputs of both untreated and treated wastewaters increased nutrient concentrations and changed physicochemical variables, with significant impacts to Monjolinho River. Seasonal variations in the measured values were also observed, in agreement with the pluviometric indexes of the region. Univariate analyses suggested no effect of the WWTP for most variables, with continued impacts at sites downstream, but non-parametric multivariate analysis indicated that these sites were recovering to chemical characteristics similar to upstream sites, apparently due to autodepuration. Therefore, multivariate methods that allow rigorous tests of multifactor hypotheses can greatly contribute to determine effects of both point and non-point sources in river systems, thus contributing to freshwater monitoring and management.     

Possible involvement of the glycocalyx in the ichthyotoxicity of Chattonella marina (Raphidophyceae): immunological approach using antiserum against cell surface structures of the flagellate

Although the ichthyotoxic mechanism of Chattonella marina is still unknown, several lines of evidence suggest that the reactive oxygen species (ROS), such as superoxide anion (O₂^-), hydrogen peroxide (H₂O₂)_, and hydroxyl radical (·OH), produced by C. marina are involved in the mortality of fish exposed to this flagellate. Recently, we found that the cell-free supernatant prepared from C. marina, which is considered to contain the glycocalyx, showed NADPH-dependent O₂^- generation. In this study, we prepared antiserum against the crude glycocalyx of C. marina. Using indirect immunofluorescence, it was confirmed that the antiserum specifically reacted with C. marina cells. In addition to C. marina, the antiserum also reacted with other raphidophycean flagellates such as Heterosigma akashiwo, Olisthodiscus luteus, and Fibrocapsa japonica, whereas no reactivity was observed against six other flagellate species tested. These results suggest that raphidophycean flagellates have common epitopes recognized by the antiserum. Interestingly, immunohistochemical analysis of paraformaldehyde-fixed gill lamellae from yellowtail exposed to C. marina revealed that the antiserum stained the surface of gill lamellae, while no such staining pattern was observed in control gill lamellae. These results suggest that the glycocalyx may be discharged when C. marina cells are inhaled into the fishes' mouths and then come into contact with the gill surface. Based on the present results, together with our previous findings, we propose that continuous accumulation of the discharged glycocalyx on the gill surface occurs during C. marina exposure, which may be responsible for the ROS-mediated severe gill tissue damage leading to fish death.     

Lectin-induced enhancement of superoxide anion production by red tide phytoplankton

Chattonella marina, a raphidophycean flagellate, is a highly toxic red tide phytoplankton which causes severe damage to fish farming. Recent studies demonstrated that Chattonella spp. continuously release superoxide anions (O₂ ⁻) while they are living. Heterosigma akashiwo, another raphidophycean flagellate, also produces O₂ ⁻. In the present study, we found that lectins such as concanavalin A (Con A), wheat germ agglutinin (WGA), and castor bean hemagglutinin (CBH) stimulated  C. marina and H. akashiwo to generate enhanced amounts of O₂ ⁻ in a concentration-dependent manner. The lectin-specific sugars potently inhibited the lectin-induced increase of O₂ ⁻ production, suggesting that the effects of lectins are mediated mainly through the interaction of these lectins with carbohydrate moiety present on the flagellate cell surface. In contrast to the potent ability of native Con A (tetravalent), succinylated Con A (divalent) showed only a slight stimulative effect on these flagellates. O₂ ⁻ production was totally inhibited by treatment with proteinase K for 30 min, without affecting the viabilities of flagellates. These results suggest that cell-surface redox enzymes may be involved in O₂ ⁻ production, and such enzymes are responsible for the lectin-stimulation. Received: 21 August 1997 / Accepted: 8 January 1998     

α1-Microglobulin levels and correlation with cadmium and other metals in urine of non-smoking women among general populations in Japan

The present study was initiated to examine urinary α₁-microglobulin (α₁-MG-U) levels among non-smoking women in the general population in Japan. A previously established database on spot urine samples from adult woman volunteers in 10 non-polluted areas all over Japan was re-examined. The data examined were on α₁-MG-U, cadmium, calcium, magnesium and zinc levels in urine (Cd-U, Ca-U, Mg-U and Zn-U, respectively), urinary creatinine (CR or cr), urine specific gravity (SG or sg), smoking habits and age. Thus, 8975 never-smoking women were selected for statistical analyses. The grand geometric mean (GM) for α₁-MG-U among the population was 2.1?mg/L or 2.5?mg/g?cr, depending on the correction for urine density. It was 1.1?µg/L or 1.3?µg/g?cr for GM Cd-U. The inter-area difference in α₁-MG-U was <1.5?mg/g?cr or <0.7?mg/L; the area with the highest or lowest GM Cd-U was not always highest or lowest in GM α₁-MG-U. The correlation coefficient (0.53) between log?Cd-U and log?α₁-MG-U (both without urine density correction) became substantially smaller when the analyte levels were corrected for CR (0.25) or SG (0.26). In multiple regression analysis, the power of influence of the five independent variables (log?Cd-U, Ca-U, Mg-U, Zn-U and age) in combination was small (R ²?≦?0.13). In contrast, logistic regression analysis suggested that α₁-MG-U might be elevated as a function of an increase in Cd-U, depending on the cut-off values. Discussion was made on dose (Cd-U_cr) and response (α₁-MG-U_cr) relationship based on information available in literatures to show that the increment in α₁-MG-U_cr per Cd-U_cr was much greater when Cd-U_cr was large, e.g., in excess of 10?µg/g?cr.     

Separation of plastic mixtures using liquid-fluidized bed technology

Separation of heavier-than-water plastic mixtures had been investigated via the fluidization of their packed beds induced by an upward flow of water. The samples examined were resin pellets and crushed plastic products including PVC, PET and PBT. On the onset of a flow, a mixed bed was swelled to the state of fluidization and separated into layers of respective resins depending on their density. The effects of the flow rate, an amount of the samples and their density difference were examined on the separation of resin pellets. Under an appropriate condition, the process was completed within a few minutes, and satisfactory separation was attained when the density difference of the samples exceeded 0.05 g cm-3. By using a column equipped with several sample outlets sorted resins could be recovered with their purity intact by withdrawing each layer successively from above through a suitable outlet. Multi-stage separation was also found to be effective in treating close-density samples. Compared with uniform-sized resin pellets, size distribution of crushed plastic samples deteriorated the separation to some extent.     

Variation in lipid classes during the molting cycle of the prawn Penaeus japonicus

The variation in the concentration and fatty acid composition of lipid classes during the molting cycle of the prawn Penaeus japonicus was investigated. The lipid concentration of the whole body reached a maximum at mid-premolt (Stage D₂) and then decreased to low level at late premolt (Stage D_3–4). The accumulation of lipids during the premolt period seemed to be attributable to the increase of both polar and neutral lipids. The increase of neutral lipids at Stage D₂ was derived from not only triglycerides but also free sterols and free fatty acids. Regarding the fatty acid composition of every lipid class, a marked variation occurred mainly at the intermolt (Stage C). In this stage, the polar lipids were rich in monoenoic acids such as 18:1 and poor in polyenoic acids such as 20:53 and 22:63. The triglycerides were rich in polyenoic acids at Stage C, but poor in monoenoic acids such as 16:1 and 18:1. The steryl esters contained large amounts of saturated acids such as 16:0 and 18:0 throughout the molting cycle, however the level of polyenoic acids increased at Stage C.     

Triterpenoid and caffeic acid derivatives in the leaves of ragweed, <Emphasis Type="Italic">Ambrosia artemisiifolia</Emphasis> L. (Asterales: Asteraceae), as feeding stimulants of <Emphasis Type="Italic">Ophraella communa</Emphasis> LeSage (Coleoptera: Chrysomelidae)

Summary. The leaf beetle Ophraella communa infests almost exclusively Ambrosia artemisiifolia in the fields of Japan, even though it normally feeds on several Asteraceous plants. A filter paper bioassay showed that the feeding of O. communa is strongly stimulated by methanolic extracts of A. artemisiifolia. The feeding stimulants for O. communa have been isolated from methanolic extracts of A. artemisiifolia. -Amyrin acetate, -amyrin acetate, 5-caffeoylquinic acid (chlorogenic acid) and 3,5-dicaffeoylquinic acid from A. artemisiifolia have been identified as feeding stimulants for O. communa. Triterpenoid derivatives (-amyrin acetate or -amyrin acetate) and caffeic acid derivatives (3, 5-dicaffoylquinic acid or 5-caffeoylquinic acid) showed feeding stimulant activity when mixed together.     

Effective diffusivity of the uranyl ion in a granite from Inada, Ibaraki, Japan

The effective diffusivity of uranium(VI) in Inada granite has been determined by through-diffusion. Experiments were performed at room temperature (20–25°C) in a 0.1 mol 1⁻¹ KCl solution where uranium is present predominantly as the poorly sorbing UO₂²⁺. An effective diffusivity (D_e) of (3.6 ± 1.6) × 10⁻¹⁴ m² s⁻¹ was obtained, close to that for uranine (nonsorbing organic tracer), but one order of magnitude lower than those obtained for Sr²⁺ and NpO₂⁺, and two orders of magnitude lower than that obtained for I⁻. According to well established theory, a proportional relationship exists between D_e and the diffusivity in the bulk of the solution (D_v). The effective diffusivity obtained in granite was not proportional to D_v. This agrees with results obtained for effective diffusivity in a Swedish granite. The ratio D_e/D_v was found to be not constant but increased with D_e or D_v. This result suggests a limit to the application of the theory.