Jui-Pin Wu

Bioaccumulation and Toxicodynamics of Cadmium to Freshwater Planarian and the Protective Effect of N-Acetylcysteine

Although toxic responses of freshwater planarians after exposure to environmental toxicants can be observed through external toxicological end points, physiological responses inside the bodies of treated planarians have rarely been investigated. The present study was designed, using cadmium (Cd) as a reference toxicant, to determine its bioaccumulation and toxicodynamics in the freshwater planarian, Dugesia japonica, after acute toxicity was obtained. Accumulated Cd concentrations, metallothionein levels, and the oxidative status in planarians were determined after exposure to Cd. Furthermore, we hypothesized that the acute death of Cd-treated planarians was associated with increased oxidative stress. After Cd-treated planarians were coexposed to antioxidant, N-acetylcysteine (NAC), we found that NAC protected planarians from Cd lethality by maintaining the oxidative status and decreasing the bioaccumulation of Cd. The results of the present study support planarians being used as a practical model for toxicological studies of environmental contaminants in the future.

Disturbances to neurotransmitter levels and their metabolic enzyme activity in a freshwater planarian exposed to cadmium

Using specific neurobehaviors as endpoints, previous studies suggested that planarian neurotransmission systems could be targets of Cd neurotoxicity. However, direct evidence for disturbed neurotransmission systems by Cd in treated planarians is still lacking. In planarians, dopamine (DA) and serotonin (5-HT) play critical roles in neuromuscular function, but little is known about their metabolic degradation. Therefore, in this study, we attempted to determine the appearances of DA, 5-HT, and their metabolic products in the freshwater planarian Dugesia japonica, characterize the activity of enzymes involved in their metabolism, and investigate the effects of Cd on planarian 5-HTergic and DAergic neurotransmission systems. Only DA, 5-HT, and 5-hydroxyindole-3-acetic acid (5-HIAA) were found in planarian tissues. Further enzymatic study revealed the activity of planarian monoamine oxidase (MAO) but not catechol-O-methyl transferase (COMT). These findings suggest that planarian MAO catalyzes the metabolism of 5-HT into 5-HIAA. However, DA metabolites from the MAO-involved metabolic pathway were not found, which might be due to a lack of COMT activity. Finally, in Cd-treated planarians, tissue levels of 5-HT and DA were decreased and MAO activity altered, suggesting that planarian neurotransmission systems are disturbed following Cd treatment.

In vivo and in vitro metabolism of tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), by the freshwater planarian, Dugesia japonica

Cigarette smoke is a risk factor for human health, and many studies were conducted to investigate its adverse effects on humans and other mammals. However, since large amounts of cigarette products are produced and consumed, it is possible that tobacco chemicals can end up in aquatic environments through several routes, thus influencing aquatic organisms. In this study, the presence of tobacco-specific nitrosamine (TSNA), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), in aquatic environment was demonstrated. Since toxic effects on and distribution patterns of tobacco chemicals in aquatic organisms were rarely studied, after results of an acute toxicity pretest were obtained, experiment was conducted to investigate the bioaccumulation pattern of NNK and distribution patterns of its metabolites, mainly 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), in NNK-treated freshwater planarians, Dugesia japonica. Results from in vivo and in vitro studies showed that NNK was readily converted to NNAL through the carbonyl reduction in bodies of NNK-treated planarians. Tissue concentrations of both chemicals increased in time- and dose-dependent manners. Furthermore, we examined the end products of NNK/NNAL α-hydroxylation in NNK-treated planarians, but only 1-(3-pyridyl)-1,4-butanediol was detected, suggesting that NNK metabolism in planarians partially differs from that in mammalian systems. This is the first report on NNK metabolism in an aquatic organism and can be used as a foundation for developing freshwater planarians as a new in vivo model for the study of NNK toxicology in the future.

Inhibitory effects of pain relief drugs on neurological enzymes: implications on their potential neurotoxicity to aquatic animals.

Pain relief medications commonly occur in the aquatic environment at measurable levels. While the neurotoxicity of pain relievers to higher vertebrates is currently known, little is known about their effects on aquatic animals. This study investigated the neurotoxicity of pain relievers to aquatic animals. We used three neurological enzymes, cholinesterase (ChE), adenosine triphosphatase (ATPase), and monoamine oxidase (MAO), from a freshwater planarian (Dugesia japonica) and green neon shrimp (Neocaridina denticulata) as biomarkers to examine the effects of pain relievers on in vitro activity. The activity of MAO and ChE, but not ATPase, was significantly inhibited by acetaminophen, but not by other pain relievers examined. It was likely that the inhibitory effects of acetaminophen on shrimp neurological enzymes were more severe than on the planarian. These findings suggest that acetaminophen is potentially neurotoxic to aquatic animals, at least in terms of neurotransmission disturbance.

Low uptakes of Cd, Cu, and Zn in Dugesia japonica, a freshwater planarian with higher tolerance to metals

ABSTRACT Although a freshwater planarian is proposed as a potential model for studying toxicities of environmental pollutants, they are less sensitive to metal exposures, compared to other freshwater invertebrates. We hypothesised that the metal uptake is low in treated planarians, which may lead to their higher tolerance to metals. The acute toxicities of Cu and Zn to Dugesia japonica were determined. The 24- and 48-h median lethal concentrations were 4.50 and 3.46 mg L−1, respectively, for Cu, and 41.97 and 37.79 mg L−1, respectively, for Zn. Accordingly, combining with the data on Cd in our previous study, higher tolerance of planarians to metals was revealed. Also, tissue levels of Cd, Cu, and Zn in treated D. japonica were determined, and their dissolved uptake rate constants (k1) were calculated. Results showed that bioaccumulations of these metals in D. japonica were, indeed, lower than that in other freshwater invertebrates having higher susceptibilities to these metals.

Ovarian Development in the Freshwater Prawn Macrobrachium asperulum (Decapoda: Palaemonidae)

Abstract Macrobrachium asperulum is a native freshwater prawn species in Taiwan, and is an ideal model for studying the reproductive biology of freshwater decapods. Also, this species may be a potential aquaculture species. This study was conducted to explore the ovarian development of female M. asperulum, as an important foundation for the knowledge of its reproductive biology. Specimens were collected in four seasons throughout the year to investigate the ovarian cycle and germ cell development, by examining the external morphology and histological structure of the ovaries. The breeding period of M. asperulum is probably from March to September each year. Developing ovaries and germ cells were characterized throughout the entire ovarian cycle. Differentiating germ cells were classified mainly into oögonia, primary oöcytes, secondary oöcytes, and mature oöcytes. Among the four major germ cells, secondary oöcytes were further divided into five types. Summarizing the above examinations and based on the relative frequency of germ cell types, six stages of ovarian cycle were identified, and this prawn is proposed as being a multiple spawning species. Observations of the present study provide important information on the reproductive biology of crustaceans and decapods in general, and also can serve as a significant foundation for the aquaculture of freshwater prawn species.