Jill Man Ying Chiu

Tissue-specific transcriptome assemblies of the marine medaka Oryzias melastigma and comparative analysis with the freshwater medaka Oryzias latipes

BackgroundThe marine medaka Oryzias melastigma has been demonstrated as a novel model for marine ecotoxicological studies. However, the lack of genome and transcriptome reference has largely restricted the use of O. melastigma in the assessment of in vivo molecular responses to environmental stresses and the analysis of biological toxicity in the marine environment. Although O. melastigma is believed to be phylogenetically closely related to Oryzias latipes, the divergence between these two species is still largely unknown. Using Illumina high-throughput RNA sequencing followed by de novo assembly and comprehensive gene annotation, we provided transcriptomic resources for the brain, liver, ovary and testis of O. melastigma. We also investigated the possible extent of divergence between O. melastigma and O. latipes at the transcriptome level.ResultsMore than 14,000 transcripts across brain, liver, ovary and testis in marine medaka were annotated, of which 5880 transcripts were orthologous between O. melastigma and O. latipes. Tissue-enriched genes were identified in O. melastigma, and Gene Ontology analysis demonstrated the functional specificity of the annotated genes in respective tissue. Lastly, the identification of marine medaka-enriched transcripts suggested the necessity of generating transcriptome dataset of O. melastigma.ConclusionsOrthologous transcripts between O. melastigma and O. latipes, tissue-enriched genes and O. melastigma-enriched transcripts were identified. Genome-wide expression studies of marine medaka require an assembled transcriptome, and this sequencing effort has generated a valuable resource of coding DNA for a non-model species. This transcriptome resource will aid future studies assessing in vivo molecular responses to environmental stresses and those analyzing biological toxicity in the marine environment.

Hypoxia causes transgenerational impairments in reproduction of fish

Hypoxia is amongst the most widespread and pressing problems in aquatic environments. Here we demonstrate that fish (Oryzias melastigma) exposed to hypoxia show reproductive impairments (retarded gonad development, decrease in sperm count and sperm motility) in F1 and F2 generations despite these progenies (and their germ cells) having never been exposed to hypoxia. We further show that the observed transgenerational reproductive impairments are associated with a differential methylation pattern of specific genes in sperm of both F0 and F2 coupled with relevant transcriptomic and proteomic alterations, which may impair spermatogenesis. The discovered transgenerational and epigenetic effects suggest that hypoxia might pose a dramatic and long-lasting threat to the sustainability of fish populations. Because the genes regulating spermatogenesis and epigenetic modifications are highly conserved among vertebrates, these results may also shed light on the potential transgenerational effects of hypoxia on other vertebrates, including humans.

Qualitative and quantitative changes in marine biofilms as a function of temperature and salinity in summer and winter

This study examined quantitative (dry weight, chlorophyll a content and C:N ratio) and qualitative (community compositions of bacteria and diatoms) changes in marine biofilms as a function of season (summer 2003 and winter 2004), temperature (16, 23 and 30 ◦ C) and salinity (20‰ ,2 7‰ and 34‰) underlaboratoryconditions.Biofilmswereallowedtodevelopfor20daysinthe laboratory,usingnaturalseawatercollectedfromPortShelter,HongKong.The following results were obtained: (1) biofilm dry weight was greater in summer than in winter, and greater at 34‰ than at 20‰; (2) biofilm chlorophyll a content was affected by all three factors (season, temperature and salinity), with significant interactive effects among the three factors; and (3) C:N ratio was affected by season (winter >summer) and temperature (30 ◦ C >16 ◦ Ci n summer), but not by salinity. Bacterial community composition was analyzed by terminal restriction-fragment length polymorphism of polymerase chain reaction-amplified 16S rRNA genes. In summer, community compositions of both bacteria and diatoms were strongly affected by salinity. In addition, natural summer biofilms that developed at three field sites, where different salinities were found, harbored appreciably different bacterial and diatom community compositions. In contrast, in winter, temperature exerted a major influence on community compositions. The present study adds to the growing evidence that environmental factors are important determinants of both the quality and quantity of marine biofilms.

Chronic Effects of Coated Silver Nanoparticles on Marine Invertebrate Larvae: A Proof of Concept Study

Silver nanoparticles (AgNPs), owing to their unique physical and chemical properties, have become increasingly popular in consumer products. However, data on their potential biological effects on marine organisms, especially invertebrates, remain very limited. This proof of principle study reports the chronic sub-lethal toxicity of two coated AgNPs (oleic acid coated AgNPs and polyvinylpyrrolidone coated AgNPs) on marine benthic invertebrate larvae across three phyla (i.e., the barnacle Balanus Amphitrite, the slipper-limpet Crepidula onyx, and the polychaete Hydroides elegans) in terms of growth, development, and metamorphosis. Bioaccumulation and biodistribution of silver were also investigated. Larvae were also exposed to silver nitrate (AgNO3) in parallel to distinguish the toxic effects derived from nano-silver and the aqueous form of silver. The sub-lethal effect of chronic exposure to coated AgNPs resulted in a significant retardation in growth and development, and reduction of larval settlement rate. The larval settlement rate of H. elegans was significantly lower in the coated AgNP treatment than the AgNO3 treatment, suggesting that the toxicity of coated AgNPs might not be solely evoked by the release of silver ions (Ag+) in the test medium. The three species accumulated silver effectively from coated AgNPs as well as AgNO3, and coated AgNPs were observed in the vacuoles of epithelial cell in the digestive tract of C. onyx. Types of surface coatings did not affect the sub-lethal toxicity of AgNPs. This study demonstrated that coated AgNPs exerted toxic effects in a species-specific manner, and their exposure might allow bioaccumulation of silver, and affect growth, development, and settlement of marine invertebrate larvae. This study also highlighted the possibility that coated AgNPs could be taken up through diet and the toxicity of coated AgNPs might be mediated through toxic Ag+ as well as the novel modalities of coated AgNPs.

Hypoxia turns genotypic female medaka fish into phenotypic males

Hypoxia caused by eutrophication is amongst the most pressing global problems in aquatic systems. Notably, more than 400 “dead zones” have been identified worldwide, resulting in large scale collapse of fisheries and major changes in the structure and trophodynamics. Recent studies further discovered that hypoxia can also disrupt sex hormone metabolism and alter the sexual differentiation of fish, resulting in male biased F1 generations and therefore posing a threat to the sustainability of natural populations. However, it is not known whether, and if so how, hypoxia can also change the sex ratio in vertebrates that have sex-determining XX/XY chromosomes. Using the Japanese medaka (Oryzias latipes) as a model, we demonstrate, for the first time, that hypoxia can turn genotypic female fish with XX chromosomes into phenotypic males. Over half of the XX females exposed to hypoxia exhibit male secondary sexual characteristics and develop testis instead of ovary. We further revealed that hypoxia can: (a) down-regulate the vasa gene, which controls proliferation of primordial germ cells and gonadal sex differentiation into ovary, and (b) up-regulate the DMY gene which resides at the sex-determining locus of the Y chromosome, and direct testis differentiation. This is the first report that hypoxia can directly act on genes that regulate sex determination and differentiation, thereby turning genotypic females into phenotypic males and leading to a male-dominant F1 population.

Polybrominated diphenyl ethers (PBDEs) alter larval settlement of marine intertidal organisms across three phyla via reducing bacterial abundance on the biofilms.

Polybrominated diphenyl ethers (PBDEs) have been widely used as flame retardants over the last three decades, and are now ubiquitous in the marine environment. While the harmful effects of PBDEs on the abnormal development and reproductive impairment in mammals and fish are well documented, the effects on marine invertebrates remain virtually unknown. Using three model intertidal species accross three phyla, including the polychaete Hydroides elegans (Phylum Annelida), the gastropod Crepidula onyx (Phylum Mollusca), and the barnacle Balanus amphitrite (Phylum Arthopoda), this study demonstrated that (a) chronic exposure to BDE-47 (at spiking concentrations up to 1000 ng L(-1)) throughout the entire larval stage did not affect settlement, development or growth of all three species per se, despite bioaccumulation was clearly evident (measured body burden ranging from approximately 7000 to 13 000 ng BDE-47 g(-1) lipid), and (b) BDE-47, at measured concentrations of 15 and 113 ng g(-1) lipid, reduced the bacterial abundance in biofilms and resulted in a concomitant change in larval settlement pattern of all the model intertidal species across three phyla.

Respiration rate and swimming activity of larvae of two sub-tidal nassariid gastropods under reduced oxygen levels: implications for their distributions in Hong Kong waters

The effects of hypoxia on the larvae of two sub-tidal nassariid gastropods, Nassarius siquijorensis and N. conoidalis were compared so as to understand how the species-specific tolerance to hypoxia might have resulted in changes in the abundance and distribution of these two species in the hypoxic Tolo Harbour, Hong Kong, since the 1980s. Respiration rates of N. siquijorensis and N. conoidalis larvae were reduced at 4.5 mg O2 l(-1), or below, as compared with the normoxic control. Significant reduction in swimming velocity was also observed for 10-day old larvae which were exposed to <2.0 mg O2 l(-1) for N. siquijorensis and <1.0 mg O2 l(-1) for N. conoidalis. The 48 h LC50 values of N. siquijorensis and N. conoidalis larvae were 0.7 and 1.7 mg O2 l(-1), respectively. The results suggested that N. siquijorensis are more tolerant to hypoxia than N. conoidalis.

Heavy metal contamination along the China coastline: A comprehensive study using Artificial Mussels and native mussels

A comprehensive study was carried out to assess metal contamination in five cities spanning from temperate to tropical environment along the coastal line of China with different hydrographical conditions. At each of the five cities, Artificial Mussels (AM) were deployed together with a native species of mussel at a control site and a polluted site. High levels of Cr, Cu and Hg were found in Qingdao, high level of Cd, Hg and Pb was found in Shanghai, and high level of Zn was found in Dalian. Furthermore, level of Cu contamination in all the five cities was consistently much higher than those reported in similar studies in other countries (e.g., Australia, Portugal, Scotland, Iceland, Korea, South Africa and Bangladesh). Levels of individual metal species in the AM showed a highly significant correlation with that in the native mussels (except for Zn in Mytilus edulis and Cd in Perna viridis), while no significant difference can be found between the regression relationships of metal in the AM and each of the two native mussel species. The results demonstrated that AM can provide a reliable time-integrated estimate of metal concentration in contrasting environments over large biogeographic areas and different hydrographic conditions, and overcome the shortcomings of monitoring metals in water, sediment and the use of biomonitors.

Diverse metabolic and stress-tolerance pathways in chasmoendolithic and soil communities of Miers Valley, McMurdo Dry Valleys, Antarctica

The majority of biomass in the McMurdo Dry Valleys of Antarctica occurs within rocks and soils, but despite the wealth of biodiversity data very little is known about the potential functionality of communities within these substrates. The putative physiological capacity of microbial communities in granite boulders (chasmoendoliths) and soils of a maritime-influenced Antarctic Dry Valleys were interrogated using the GeoChip microarray. Diversity estimates revealed surprisingly high diversity and evenness in both communities, with Chlorobi and Deinococci in soils accounting for major differences between the substrates. Autotrophs were more diverse in chasmoendoliths, and diazotrophs more diverse in soils. Both substrates revealed a previously unappreciated abundance of Halobacteria (Archaea), Ascomycota (Fungi) and Basidiomycoyta (Fungi). The fungi accounted for much of the differences between substrates in metabolic pathways associated with carbon transformations, particularly for aromatic compounds. Nitrogen fixation genes were more common in soils, although nitrogen catabolism genes were abundant in chasmoendoliths. Stress response pathways were more diverse in chasmoendoliths, possibly reflecting greater environmental stress in this exposed substrate compared with subsurface soils. Overall diversity of stress-tolerance genes was markedly lower than that recorded for inland locations where environmental stress is exacerbated. We postulate that the chasmoendolithic community occupies a key role in biogeochemical transformations in Dry Valley systems where granite substrates are abundant among open soils. The findings indicate that a substantial upward revision to estimates of biologically active surfaces in this system is warranted.

Pyrosequencing of the bacteria associated with Platygyra carnosus corals with skeletal growth anomalies reveals differences in bacterial community composition in apparently healthy and diseased tissues

Corals are rapidly declining globally due to coral diseases. Skeletal growth anomalies (SGA) or “coral tumors” are a group of coral diseases that affect coral reefs worldwide, including Hong Kong waters in the Indo-Pacific region. To better understand how bacterial communities may vary in corals with SGA, for the first time, we examined the bacterial composition associated with the apparently healthy and the diseased tissues of SGA-affected Platgyra carnosus using 16S ribosomal rRNA gene pyrosequencing. Taxonomic analysis revealed Proteobacteria, Bacteroidetes, Cyanobacteria, and Actinobacteria as the main phyla in both the apparently healthy and the diseased tissues. A significant difference in the bacterial community composition was observed between the two conditions at the OTU level. Diseased tissues were associated with higher abundances of Acidobacteria and Gemmatimonadetes, and a lower abundance of Spirochaetes. Several OTUs belonging to Rhodobacteraceae, Rhizobiales, Gammaproteobacteria, and Cytophaga-Flavobacterium-Bacteroidetes (CFB) were strongly associated with the diseased tissues. These groups of bacteria may contain potential pathogens involved with the development of SGA or opportunistic secondary or tertiary colonizers that proliferated upon the health-compromised coral host. We suggest that these bacterial groups to be further studied based on inoculation experiments and testing of Kochs postulates in efforts to understand the etiology and progression of SGA.