Phytochelatins do not correlate with the level of Cd accumulation in Chlamydomonas spp

Chlamydomonas acidophila KT-1 and Chlamydomonas acidophila DVB238 exhibit a strong heavy metal tolerance, but C. acidophila DVB238 can accumulate a much higher amount of Cadmium (Cd) than C. acidophila KT-1. Phytochelatins (PCs) are known to play an important role in the detoxification of several toxic heavy metals, but the relationship between PCs and Cd accumulation is not clear. PC metabolism and Cd accumulation were investigated by using three Chlamydomonas strains including Chlamydomonas reinhardtii C-9 as a standard alga. The results showed that the PC content did not correlate closely with the level of Cd accumulation, maintenance of a high GSH level seeming to be more important for Cd accumulation. The ultrastructure of C. acidophila KT-1 was extremely disrupted by a great increase in starch granules, which resulted in a moribund state, but hyper-accumulator C. acidophila DVB238 did not exhibit an increase in starch granules in its cells, in spite of Cd accumulation in its chloroplasts, cytosol and vacuoles. These results indicated that C. acidophila DVB238 probably has a developed detoxification system preventing such as destruction of the cells due to Cd toxicity.     

Sulfur isotopic variations in soft tissues of five benthic animals from the reductive,tidal-flat sediments in northern Kyushu,Japan

The sulfur nutrition of five benthic animals living in or on reductive, tidal-flat sediments has been studied by using the sulfur isotopic signatures of their soft tissues. The mean '³⁴S values (+6.1‰ to +13.8‰, relative to the Cañyon Diablo troilite reference) of these animals' tissues are lower than those of many common marine animals, which have values close to those of seawater sulfate-sulfur (+21‰). This indicates that these animals use a ³⁴S-depleted sulfur source, which may be derived from bacterial sulfide in the sediments (less than -20‰). The animals that inhabit such sediments are adapted to an anoxic environment, where toxic hydrogen sulfide prevails. Due to the sulfide-rich habitat, benthic animals are expected to assimilate hydrogen sulfide directly during detoxification, or to prefer a diet which has a low '³⁴S signature. Variations in '³⁴S values of the sampled molluscs were fairly small, whereas migratory shrimp and fish tended to have larger variations, implying that migratory animals feed on diverse prey, while the molluscs feed on a common food, or their assimilative capacity for sulfide-sulfur does not vary among individuals. A gastropod inhabiting the surface of the reductive sediment showed a distinctively low '³⁴S value (+5.8‰ to +6.4‰), indicating that the animal fed on detritus from reeds and halophytes with low '³⁴S values (-19.1‰ to -2.1‰). This study suggests that some benthic animals incorporate sulfide-sulfur for their nutrition and that sulfur isotope composition can be a useful indicator in the analysis of the food habitat of animals.     

Reproductive disturbance of Japanese bumblebees by the introduced European bumblebee Bombus terrestris

The European bumblebee, Bombus terrestris, is an invasive eusocial species whose distribution is expanding greatly beyond its native range because numerous colonies are imported to or locally produced in non-native countries for pollination of agricultural crops. Closely related species exist in Japan where the unrestricted import and use of B. terrestris has resulted in the establishment of wild colonies. Laboratory studies previously showed that B. terrestris and Japanese native species can copulate and produce fertilized eggs. Although these eggs do not hatch, the interspecific mating can cause a serious reproductive disturbance to native bumblebees. In this study, we determined the frequencies of interspecies mating between B. terrestris males and native bumblebee queens in the wild on the islands of Hokkaido and Honshu by analyzing the DNA sequences of spermatozoa stored in spermathecae of native queens. We found that 20.2% of B. hypocrita hypocrita queens and 30.2% of B. hypocrita sapporoensis queens had spermatozoa of B. terrestris males in their spermathecae. Given that a Bombus queen generally mates only once in her life, such high frequencies of interspecific mating with B. terrestris pose serious threats to the populations of native bumblebees in Japan.     

Hydrologic connection between ponds positively affects macrophyte alpha and gamma diversity but negatively affects beta diversity

Connections between habitat patches can positively influence the number of species in respective patches, providing a basis for preferentially conserving interconnected patches. However, from a regional perspective, it is not known whether conserving multiple sets of interconnected habitat patches would include more species (i.e., show higher gamma diversity) than conserving multiple, unconnected, solitary patches. We studied aquatic macrophytes in 15 sets of unidirectionally interconnected ponds and 19 unconnected ponds and also tested whether alpha and beta diversity, expressed as the number of species and dissimilarity in species composition, respectively, differed between connected and unconnected ponds. We found that gamma diversity was higher in connected ponds than in unconnected ponds, even after controlling for surface area. This resulted from a higher alpha diversity in connected ponds, despite lower beta diversity. These results suggest that connections between habitat patches positively influence diversity at both local and regional scales. When the total surface area available for conservation is limited, interconnected habitat patches should be preferentially conserved.     

Variation of zinc and copper in metallothionein-like protein in killifish (Oryzias latipes) exposed to cadmium

Killifish (Oryzias latipes) were exposed for 0, 24, 48 and 168 hours to cadmium solution. The hepatic cytoplasm was fractionated by using Sephadex G-75 and the content of zinc, copper and cadmium in the fractions was measured. Cadmium content in the metallothionein fraction increased with increase of exposure time. Zinc content in the metallothionein fraction was reduced at early exposure time and increased after prolonged exposure. Copper content in the metallothionein fraction was unchanged. It is shown that zinc bound to thionein in the liver is partially replaced by cadmium and that at least two isomers of metallothionein occur in this fish.     

Polychlorinated dibenzo-p-dioxins, -dibenzofurans, and biphenyls in domestic animal food stuff and their fat

Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzo-p-dioxins (PCDFs) and dioxin-like coplanar PCBs and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs) were measured in domestic animal related samples such as feed ingredient, mixed feed and animal fat from Japan. The mean concentration of total PCDDs, PCDFs and coplanar PCBs in fish oil was 17000pg g-1 lipid wt, which 9- to 30-fold greater than that from other animal originated ingredients. Mixed feed for chicken and chicken fat contained greater total PCDDs, PCDFs, coplanar PCBs concentrations (190 and 2100pg g-1 lipid wt) compared to those for pig (120 and 320pg g-1 lipid wt). WHO-TEQs were ranged 0.39-13, 0.15-0.22 and 0.73-2.8pg g-1 lipid wt for ingredients, mixed feeds and animal fats, respectively. Greater biomagnification factor was noticed for congeners 1,2,3,7,8-PeCDD, 1,2,3,6,7,8-HxCDD, 2,3,7,8-TCDF and 1,2,3,4,7,8-HxCDF in chicken and pig fat. Biomagnification factors for both mono- and non-ortho coplanar PCBs were greater in chicken fat compared to those in pig fat.     

Biochemical and cellular effects of electrophiles present in ambient air samples

Ambient vapor-phase samples collected in Riverside, California had shown that both redox and electrophilic activity were present, with the vapor phase containing higher levels of electrophiles than the particle phase. In this study, the biochemical effects of the vapor-phase electrophiles were examined using the purified thiol proteins, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), protein tyrosine phosphatase 1B (PTP1B) and KELCH-1 like ECH-associated protein 1 (Keap1). The results demonstrated that the thiol proteins were inactivated by the vapor-phase samples through covalent modifications. Next, two cellular responses, epidermal growth factor receptor (EGFR)/mitogen-activated protein (MAP) kinase and NF-E2-related factor 2 (Nrf2), to the ambient vapor-phase samples were assessed in A549 and RAW 264.7 cell lines, respectively. The vapor-phase samples, at non-oxidative concentrations, increased phosphorylation of EGFR, which is negatively regulated by PTP1B, and its downstream MAP kinase, extracellular signal-regulated kinase (ERK)1/2. Activation of Nrf2, which requires Keap1 alkylation, and expression of its downstream proteins were also observed. The electrophilic compounds present in ambient vapor-phase were shown to modify cellular proteins through covalent modification and to activate diverse cellular responses that can lead to inflammatory and adaptive responses.     

Variability and reliability of POP concentrations in multiple breast milk samples collected from the same mothers

Risk assessment of infant using a realistic persistent organic pollutant (POP) exposure through breast milk is essential to devise future regulation of POPs. However, recent investigations have demonstrated that POP levels in breast milk collected from the same mother showed a wide range of variation daily and monthly. To estimate the appropriate sample size of breast milk from the same mother to obtain reliable POP concentrations, breast milk samples were collected from five mothers living in Japan from 2006 to 2012. Milk samples from each mother were collected 3 to 6 times a day through 3 to 7 days consecutively. Food samples as the duplicated method were collected from two mothers during the period of breast milk sample collection. Those were employed for POP (PCBs, DDTs, chlordanes, and HCB) analysis. PCB concentrations detected in breast milk samples showed a wide range of variation which was maximum 63 and 60% of relative standard deviation (RSD) in lipid and wet weight basis, respectively. The time course trend of those variations among the mothers did not show any typical pattern. A larger amount of PCB intake through food seemed to affect 10 h after those concentrations in breast milk in lipid weight basis. Intraclass correlation coefficient (ICC) analyses indicated that the appropriate sample size for good reproducibility of POP concentrations in breast milk required at least two samples for lipid and wet weight basis.