Waterborne and Dietborne Toxicity of Inorganic Arsenic to the Freshwater Zooplankton Daphnia magna.

Waterborne and dietborne exposure are both important sources for the accumulation of inorganic arsenic (iAs) in aquatic organisms. Although the waterborne toxicity of iAs has been extensively investigated, its dietborne toxicity has received little attention. The present study examined the acute and chronic toxicity of arsenate (iAsV) and arsenite (iAsIII) to the freshwater zooplankton species Daphnia magna under both waterborne and dietborne exposure scenarios. The bioaccumulation, speciation, and tissue and subcellular distributions of arsenic were analyzed to understand the mechanisms accounting for differences in toxicity related to different arsenic species, exposure scenarios, and exposure duration. The toxicity of iAs increased with exposure time, and iAsIII was more toxic than iAsV. Moreover, although dietborne iAs had no acute effect on D. magna, it incurred significant toxicity in the chronic-exposure experiment. Nevertheless, the toxicity of dietborne iAs was still lower than that of waterborne iAs regardless of the exposure duration. This difference was found to be caused by the lower bioaccumulation of dietborne iAs, its higher distribution in the gut and in the biologically detoxified subcellular fraction, and greater transformation to the less toxic dimethylarsinic acid. Overall, the dietborne toxicity of iAs should be considered when evaluating the environmental risks posed by arsenic.

New insights into the fungal community from the raw genomic sequence data of fig wasp Ceratosolen solmsi.

BACKGROUND: To date, biologists have discovered a large amount of valuable information from assembled genomes, but the abundant microbial data that is hidden in the raw genomic sequence data of plants and animals is usually ignored. In this study, the richness and composition of fungal community were determined in the raw genomic sequence data of Ceratosolen solmsi (RGSD-CS). RESULTS: To avoid the interference from sequences of C. solmsi, the unmapped raw data (about 17.1%) was obtained by excluding the assembled genome of C. solmsi from RGSD-CS. Comparing two fungal reference datasets, internal transcribed spacer (ITS) and large ribosomal subunit (LSU) of rRNA, the ITS dataset discovered a more diverse fungal community and was therefore selected as the reference dataset for evaluating the fungal community based on the unmapped raw data. The threshold of 95% sequence identity revealed many more matched fungal reads and fungal richness in the unmapped raw data than those by identities above 95%. Based on the threshold of 95% sequence identity, the fungal community of RGSD-CS was primarily composed of Saccharomycetes (88.4%) and two other classes (Agaricomycetes and Sordariomycetes, 8.3% in total). Compared with the fungal community of other reported fig wasps, Agaricomycetes and Eurotiomycetes were found to be unique to C. solmsi. In addition, the ratio of total fungal reads to RGSD-CS was estimated to be at least 4.8 × 10(-3), which indicated that a large amount of fungal data was contained in RGSD-CS. However, rarefaction measure indicated that a deeper sequencing coverage with RGSD-CS was required to discover the entire fungal community of C. solmsi. CONCLUSION: This study investigated the richness and composition of fungal community in RGSD-CS and provided new insights into the efficient study of microbial diversity using raw genomic sequence data.

TiO2 nanoparticles act as a carrier of Cd bioaccumulation in the ciliate Tetrahymena thermophila.

When nanoparticles can enter a unicellular organism directly, how may they affect the bioaccumulation and toxicity of other pollutants already present in the environment? To answer this question, we conducted experiments with a protozoan Tetrahymena thermophila. The well-dispersed polyacrylate-coated TiO2 nanoparticles (PAA-TiO2-NPs) were used as a representative nanomaterial, and Cd as a conventional pollutant. We found that PAA-TiO2-NPs could get into Tetrahymena cells directly. Such internalization was first induced by low concentrations of Cd, but later suppressed when Cd concentrations were higher than 1 µg/L. Considering its significant adsorption on PAA-TiO2-NPs, Cd could be taken up by T. thermophila in the form of free ion or metal-nanoparticle complexes. The latter route accounted for 46.3% of Cd internalization. During the 5 h depuration period, 4.34-22.1% of Cd was excreted out, which was independent of the concentrations of intracellular Cd and PAA-TiO2-NPs. On the other hand, both free and intracellular Cd concentrations only partly predicted its toxicity at different levels of PAA-TiO2-NPs. This may have resulted from PAA-TiO2-NPs' synergistic effects and the distinct subcellular distribution of Cd taken up via the two routes above. Overall, we should pay attention to the carrier effects of nanoparticles when assessing their environmental risks.

Influence of humic acid and fluvic acid on the altered toxicities of arsenite and arsenate toward two freshwater algae.

Arsenic pollution in freshwater poses a serious threat to aquatic organisms. However, dissolved organic matter (DOM) in water can modulate arsenic environmental toxicity by either suppressing or promoting its bioaccumulation. In this study, we investigated the toxicity, bioaccumulation, and biotransformation of inorganic arsenic (arsenite AsIII and arsenate AsV) combined with two types of DOM, i.e., humic acid (HA) and fulvic acid (FA), in the algae Chlamydomonas reinhardtii and Ochromonas danica. C. reinhardtii has a cell wall and cannot bioaccumulate arsenic complexation, whereas O. danica has no cell wall. Without DOM, AsV was more toxic than AsIII for C. reinhardtii, and AsV was less toxic than AsIII for O. danica. HA and FA addition reduced AsV and AsIII toxicities; the larger molecular weight (Mw) of HA contributed to the reduction in toxicity to an even greater extent, and reduced arsenic accumulation while promoting the biotransformation ability of C. reinhardtii, which has a cell wall. However, HA and FA addition increased AsV and AsIII toxicities and arsenic accumulation while relatively enhancing the biotransformation ability of O. danica, which has no cell wall. Coupling toxicity, bioaccumulation, and biotransformation, DOM (HA and FA) contributed to the altered toxicity of freshwater algae to AsV and AsIII through reduced/increased arsenic accumulation and enhanced biotransformation. Overall, our study considered the combined toxicity of inorganic arsenic and DOM in phytoplankton, helping estimate the potential environmental risk of arsenic in aqueous environments.

Arsenic bioaccumulation and biotransformation in aquatic organisms.

Arsenic exists universally in freshwater and marine environments, threatening the survival of aquatic organisms and human health. To elucidate arsenic bioaccumulation and biotransformation processes in aquatic organisms, this review evaluates the dissolved uptake, dietary assimilation, biotransformation, and elimination of arsenic in aquatic organisms and discusses the major factors influencing these processes. Environmental factors such as phosphorus concentration, pH, salinity, and dissolved organic matter influence arsenic absorption from aquatic systems, whereas ingestion rate, gut passage time, and gut environment affect the assimilation of arsenic from foodstuffs. Arsenic bioaccumulation and biotransformation mechanisms differ depending on specific arsenic species and the involved aquatic organism. Although some enzymes engaged in arsenic biotransformation are known, deciphering the complicated synthesis and degradation pathway of arsenobetaine remains a challenge. The elimination of arsenic involves many processes, such as fecal excretion, renal elimination, molting, and reproductive processes. This review facilitates our understanding of the environmental behavior and biological fate of arsenic and contributes to regulation of the environmental risk posed by arsenic pollution.

Effects of nitrogen and phosphorus on arsenite accumulation, oxidation, and toxicity in Chlamydomonas reinhardtii.

We studied arsenite (iAs(III)) accumulation, oxidation, and toxicity in the freshwater green alga Chlamydomonas reinhardtii under nutrient-enriched (+NP), phosphorus-limited (-P), and nitrogen-limited (-N) conditions. The -P alga (55.1 µM) had a Michaelis constant (Kd) for uptake approximately one tenth of the +NP (419 µM) and -N (501 µM) cells, indicating iAs(III) uptake inhibition by extracellular phosphate. This conclusion was supported by the hyperbolic reduction in iAs(III) uptake rate (V) from 9.2 to 0.8 µmol/g-dw/h when the extracellular phosphate concentration went up from 0 to 250 µM. The maximal iAs(III) uptake rate (Vmax) of the -N alga (24.3 µmol/g-dw/h) was twice as much as that of the +NP (12 µmol/g-dw/h) and -P (8.1 µmol/g-dw/h) cells. It implies that more arsenic transporters were synthesized under the -N condition. Once accumulated, iAs(III) was oxidized and a higher proportion of arsenate (iAs(V)) was observed at lower [As]dis or under nutrient-limited conditions. Nevertheless, iAs(III) oxidation mainly occurred outside the cells with the extent of oxidation reciprocal to [As]dis. Based on the logistic modeling of the concentration-response curves in the +NP, -P, and -N toxicity tests, iAs(III) had an [As]dis-based EC50 of 1763, 13.1, and 1208 µM and an intracellular arsenic concentration based EC50 of 35.6, 28.8, and 195 µmol/g-dw, respectively. Higher iAs(III) toxicity to the -P cells occured because of their increased iAs(III) accumulation, whereas the underlying mechanisms why the -N alga was more tolerant need to be further revealed. Overall, both N and P had remarkable effects on the behavior and effects of iAs(III), which cannot be disregarded in the biogeochemical cycling research of arsenic.

Toxicity and bioaccumulation kinetics of arsenate in two freshwater green algae under different phosphate regimes.

In the present study, the toxicity and bioaccumulation kinetics of arsenate in two green algae Chlamydomonas reinhardtii and Scenedesmus obliquus under phosphate-enriched (+P) and limited (-P) conditions were investigated. P-limitation was found to aggravate arsenate toxicity and S. obliquus was more tolerant than C. reinhardtii. Such phosphate-condition-dependent or algal-species-specific toxicity difference was narrowed when the relative inhibition of cell growth was plotted against intracellular arsenate content instead of its extracellular concentration. The discrepance was further reduced when the intracellular ratio of arsenic to phosphorus was applied. It suggests that both arsenate bioaccumulation and intracellular phosphorus played an important role in arsenate toxicity. On the other hand, arsenate uptake was induced by P-limitation and its variation with ambient arsenate concentration could be well fitted to the Michaelis-Menten model. Arsenate transporters of S. obliquus were found to have a higher affinity but lower capacity than those of C. reinhardtii, which explains its better regulation of arsenate accumulation than the latter species in the toxicity experiment. Further, arsenate depuration was facilitated and more was transformed to arsenite in C. reinhardtii or under -P condition. Intracellular proportion of arsenite was also increased after the algae were transferred from the long-term uptake media to a relatively clean solution in the efflux experiment. Both phenomena imply that algae especially the sensitive species could make physiological adjustments to alleviate the adverse effects of arsenate. Overall, our findings will facilitate the application of algae in arsenate remediation.

Host sex-specific parasites in a functionally dioecious fig: a preference way of adaptation to their hosts.

Host-parasites interaction is a common phenomenon in nature. Diffusive coevolution might maintain stable cooperation in a fig-fig wasps system, in which the exploiter might diversify their genotype, phenotype, or behavior as a result of competition with pollinator, whereas the figs change flower syconia, fruits thickness, and syconia structure. In functionally dioecious Ficus auriculata, male figs and female figs contain two types of florets on separate plant, and share high similarities in outside morphology. Apocryptophagus (Sycophaginae, Chalcidoidea, Hymenoptera) is one of few groups of nonpollinating fig wasps that can reproduce within both male and female figs. On the basis of the morphology and DNA barcoding, evidence from partial sequences of mitochondrial cytochrome c oxidase I and nuclear internal transcribed spacer 2, we found that there are two nonsibling Apocryptophagus species living on male and female F. auriculata figs, respectively. We estimated that these two species diverged about 19.2 million years ago. Our study suggests that the host shift from Ficus variegate or Ficus prostrata fig species to male figs is a preference way for Apocryptophagus wasps to adapt to the separation of sexual function in diecious figs. Furthermore, to escape the disadvantage or sanction impact of the host, the exploiter Apocryptophagus wasps can preferably adapt to exploiting each sex of the figs, by changing their oviposition, niche shift, and habitat.

Obligate mutualism within a host drives the extreme specialization of a fig wasp genome.

BACKGROUND: Fig pollinating wasps form obligate symbioses with their fig hosts. This mutualism arose approximately 75 million years ago. Unlike many other intimate symbioses, which involve vertical transmission of symbionts to host offspring, female fig wasps fly great distances to transfer horizontally between hosts. In contrast, male wasps are wingless and cannot disperse. Symbionts that keep intimate contact with their hosts often show genome reduction, but it is not clear if the wide dispersal of female fig wasps will counteract this general tendency. We sequenced the genome of the fig wasp Ceratosolen solmsi to address this question. RESULTS: The genome size of the fig wasp C. solmsi is typical of insects, but has undergone dramatic reductions of gene families involved in environmental sensing and detoxification. The streamlined chemosensory ability reflects the overwhelming importance of females finding trees of their only host species, Ficus hispida, during their fleeting adult lives. Despite long-distance dispersal, little need exists for detoxification or environmental protection because fig wasps spend nearly all of their lives inside a largely benign host. Analyses of transcriptomes in females and males at four key life stages reveal that the extreme anatomical sexual dimorphism of fig wasps may result from a strong bias in sex-differential gene expression. CONCLUSIONS: Our comparison of the C. solmsi genome with other insects provides new insights into the evolution of obligate mutualism. The draft genome of the fig wasp, and transcriptomic comparisons between both sexes at four different life stages, provide insights into the molecular basis for the extreme anatomical sexual dimorphism of this species.

Accumulation kinetics of arsenic in Daphnia magna under different phosphorus and food density regimes.

In the present study, the dissolved uptake, dietary assimilation, and elimination of arsenic (initially added as arsenate) in the freshwater cladoceran Daphnia magna were examined. A biphasic correlation between the arsenic uptake rate and its ambient concentration, as well as a two-saturation-site arsenic uptake competition with phosphate was observed. The calculated uptake rate constant, as influenced by the ambient phosphorus concentration, ranged from 0.035 to 0.35 L/g/d. Food concentration substantially decreased (by 23.2-64.4%) the arsenic assimilation efficiency with the incipient limiting algal food concentration of 3.86 mg/L dry weight. Arsenic assimilation by the daphnids was independent of their own phosphorus status, but was lower when their algal diet was phosphorus-limited and thus contained a higher proportion of arsenite due to the enhanced biotransformation. Arsenic efflux rate constant ranged from 0.34 to 0.44 d(-1) with increased food concentration slightly facilitating its loss. Excretion, accounting for 51.3 to 60.6% of total loss, was the dominant pathway for arsenic elimination with a remarkable contribution from offspring production (24.7-29.8%), whereas molting (3.64-4.05%) and egestion (7.9-11.9%) had minor roles only. According to the well-established biokinetic model, dietary assimilation was predicted to be the main pathway for arsenic bioaccumulation in the daphnids, and arsenic has a great potential to be biodiminished along the food chain.

Effects of microcystin-LR on the metal bioaccumulation and toxicity in Chlamydomonas reinhardtii.

Microcystin-LR (MC-LR) is one of the most notorious toxins liberated from cyanobacteria in eutrophicated freshwater ecosystems. Its effects on the bioaccumulation and toxicity of Cd(2+), CrO(4)(2-), Cu(2+), and Zn(2+) in a green alga Chlamydomonas reinhardtii were investigated in the present study. The metal bioaccumulation in the alga was unaffected by MC-LR. The surface-adsorbed and intracellular metal concentrations in the treatments with and without the addition of MC-LR could be well simulated by a single Freundlich isotherm for each metal with their accumulation ability following the order of Cu(2+) > Cd(2+) > Zn(2+) > CrO(4)(2-). The bioavailable metal concentrations measured by diffusion gradients in thin-films remained unchanged when MC-LR was applied. Accordingly, the growth of C. reinhardtii was similarly inhibited at the same metal concentration regardless of the addition of MC-LR. The metal toxicity could also be well delineated with the classic free ion activity and biotic ligand models. However, the intracellular metal concentration was found to have the best predictability suggesting its more direct relationship with metal toxicity. Metal exposure induced the accumulation of MC-LR in the alga, which was leveled off at high metal levels. The underlying uptake mechanisms need to be further examined.

Wolbachia infection and dramatic intraspecific mitochondrial DNA divergence in a fig wasp.

Mitochondria and Wolbachia are maternally inherited genomes that exhibit strong linkage disequilibrium in many organisms. We surveyed Wolbachia infections in 187 specimens of the fig wasp species, Ceratosolen solmsi, and found an infection prevalence of 89.3%. DNA sequencing of 20 individuals each from Wolbachia-infected and -uninfected subpopulations revealed extreme mtDNA divergence (up to 9.2% and 15.3% in CO1 and cytochrome b, respectively) between infected and uninfected wasps. Further, mtDNA diversity was significantly reduced within the infected group. Our sequencing of a large part of the mitochondrial genome from both Wolbachia-infected and -uninfected individuals revealed that high sequence divergence is common throughout the mitochondrial genome. These patterns suggest a partial selective sweep of mitochondria subsequent to the introduction of Wolbachia into C. solsmi, by hybrid introgression from a related species.

Molecular approaches to identify cryptic species and polymorphic species within a complex community of fig wasps.

Cryptic and polymorphic species can complicate traditional taxonomic research and both of these concerns are common in fig wasp communities. Species identification is very difficult, despite great effort and the ecological importance of fig wasps. Herein, we try to identify all chalcidoid wasp species hosted by one species of fig, using both morphological and molecular methods. We compare the efficiency of four different DNA regions and find that ITS2 is highly effective for species identification, while mitochondrial COI and Cytb regions appear less reliable, possibly due to the interference signals from either nuclear copies of mtDNA, i.e. NUMTs, or the effects of Wolbachia infections. The analyses suggest that combining multiple markers is the best choice for inferring species identifications as any one marker may be unsuitable in a given case.