Cytochrome P450 1a (Cyp1a) is an important enzyme for metabolism of organic pollutants. To understand its reaction to polycyclic aromatic hydrocarbons (PAHs), we knocked out this gene in a marine model fish, Javanese medaka, Oryzias javanicus, using the CRISPR/Cas 9 system. A homozygous mutant (KO) strain with a four-base deletion was established using an environmental DNA (eDNA)-based genotyping technique. Subsequently, KO, heterozygous mutant (HT), and wild-type (WT) fish were exposed to model pollutants, pyrene and phenanthrene, and survivorship and swimming behavior were analyzed. Compared to WT, KO fish were more sensitive to pyrene, suggesting that Cyp1a transforms pyrene into less toxic metabolites. Conversely, WT fish were sensitive to phenanthrene, suggesting that metabolites transformed by Cyp1a are more toxic than the original compound. HT fish showed intermediate results. Thus, comparative use of KO and WT fish can distinguish modes of pollutant toxicity, providing a deeper understanding of fish catabolism of environmental pollutants.

Several organisms have been used as indicators, bio-monitoring agents or test organisms in ecotoxicological studies. A close relative of the well established Japanese medaka, the Java medaka (Oryzias javanicus), has the potential to be a test organism. The fish is native to the estuaries of the Malaysian Peninsula, Thailand, Indonesia and Singapore. In this study, newly fertilised eggs were exposed to different concentrations of Cd and Hg. Observations were done on the development of the embryos. Exposure to low levels of Cd and Hg (0.01–0.05ppm) resulted in several developmental disorders that led to death. Exposure to ⩾1.0ppm Cd resulted in immediate developmental arrest. The embryos of Java medaka showed tolerance to a certain extent when exposed to ⩾1.0ppm Hg compared to Cd. Based on the sensitivity of the embryos, Java medaka is a suitable test organism for ecotoxicology in the tropical region.

This study was conducted to investigate the median lethal concentration (LC50) of copper pyrithione (CuPT) at 96-hr exposure on adult Javanese medaka (Oryzias javanicus) in revealing toxicological effects of CuPT contamination in the tropical area. Wild stock fishes were acclimatized for 14-days prior analysis. Triplicate of test tanks for seven test concentrations were placed with ten fishes each, this includes two control tanks. The behaviour of the tested fishes was manually observed through a camera. The LC50 of CuPT at 96-h was found to be 16.58mg/L. Tested fishes swam slowly in vertical movement and swam fast towards food during feeding time as the sign of stress behaviour. Meanwhile, fishes in the two control groups swam actively in a horizontal manner and no excitement during feeding time. No mortality in control groups. Results indicate CuPT to be toxic to Javanese medaka at low concentration and caused behavioural stress.

Glyphosate is globally a widely used herbicide, yet there is little information on their toxicity to marine fishes. Java medaka, a small tropical fish native to coastal areas in several Southeast Asian countries, is viewed as a suitable candidate for toxicity test and thus was used for this study. Java medaka adults were cultured in the laboratory and the fertilized eggs of the F2 generation were exposed to different concentrations of glyphosate-based herbicide (100, 200, 300, 400 and 500ppm) until they hatched. The survival and hatching rates of the embryos, changes in the heart rate and morphological impairments were recorded. Generally, survival and hatching percentage decreased as glyphosate concentration increased. Absence of pectoral fin(s) and cornea, permanently bent tail, irregular shaped abdomen, and cell disruption in the fin, head and abdomen are among the common teratogenic effects observed. Furthermore, risk factor also increased with the increased in glyphosate concentrations.

The Java medaka (Oryzias javanicus) is distributed in tropical brackish water and is considered as an ecotoxicological experimental organism for assessing diverse pollutions and global climate change effects in the ocean. In this study, we sequenced and assembled the genome of O. javanicus using the Oxford Nanopore technique and anchored the scaffolds to the 24 genetic linkage map of a sister species Oryzias melastigma. The assembled genome consisted of 773 scaffolds including 24 LG-based scaffolds, and the estimated genome length was 846.3 Mb (N50 = 19.3 Mb), containing 24,498 genes. As detoxification processes are crucial in aquatic organisms, antioxidant-related genes including glutathione S-transferases, superoxide dismutase, catalase, and glutathione peroxidase were identified in this study. In the genome of O. javanicus, a total of 21 GSTs, 4 SODs, 1 CAT, and 7 GPxs were identified and showed high similarities between sister species O. melastigma and Oryzias latipes. In addition, despite having 8 classes of cytosolic GSTs family, medaka showed no presence of GST pi and sigma classes, which are predominantly found in carp and salmon, but not in neoteleostei. This study adds another set to genome-library of Oryzias spp. and is a useful resource for better understanding of the molecular ecotoxicology.

To assess possible adverse effects of residual chlorines from hypochlorite-treated seawater to non-target marine organisms, bioassays were carried out on marine amphipod Hyale barbicornis and estuarine fish Oryzias javanicus. Acute toxicity tests were first conducted using various concentrations of sodium hypochlorite (NaOCl) followed by a long-term exposure to residual chlorines from a test water treated with 1 mg L⁻¹ NaOCl. Results showed that NaOCl was acutely toxic to both organisms. However, long-term exposure to residual chlorines from NaOCl-treated waters caused no major adverse effects to both organisms under laboratory conditions since free chlorines in the treated water was reduced to about 10% by 23-h holding and 1-h aeration. No H. barbicornis died but residual chlorine-exposed juveniles had significantly shorter body lengths at the end of exposure. Residual chlorine-exposed O. javanicus also showed no significant differences to that of the control in all measured endpoints except for hatching time. The results suggest that using 1 mg L⁻¹ NaOCl for disinfection of ballast waters will produce residual chlorines that is far below the LC50 and EC50 of H. barbicornis and O. javanicus even on a long-term basis.