Detection of phenoloxidase activity in early stages of the Pacific oyster Crassostrea gigas (Thunberg).

The presence of phenoloxidase (PO) activity was detected in different developmental stages of the Pacific oyster, Crassostrea gigas. A significant reduction in PO activity was observed from the 6h embryo stage to the day 11 larvae by spectrophotometry. A progressive increase was also observed from the day 13 larvae right through to the juvenile stage. The microscopy studies with '6h embryo' and adult samples confirmed the presence of PO activity. Various modulators of PO activity were used to study the triggering of pro-phenoloxidase (proPO) activating system of C. gigas but also to confirm the exact nature of the monitored activity. The enzyme activation mechanisms appear to differ with the developmental stage: bacterial lipopolysaccharides constitute an early elicitor of the proPO-PO system, whereas a purified trypsin triggers proPO-PO system in C. gigas spat. Phenoloxidase activity was totally suppressed by PO-specific inhibitors such as beta-2-mercaptoethanol, sodium diethyldithiocarbonate and tropolone. This study demonstrated the selective response of PO-like activity by different elicitors and suggested that proPO-PO activating system, which is supposed to play an important function in non-self recognition and host immune reactions in oyster, is expressed early in the Pacific oyster, C. gigas.

Impact of diuron on aneuploidy and hemocyte parameters in Pacific oyster, Crassostrea gigas.

Diuron is a substituted urea herbicide used for agricultural and nonagricultural weed control. Its widespread use and relatively slow breakdown led us to analyze its influence on aneuploidy level (lowered chromosome number in a percentage of somatic cells) and hemocyte parameters in Pacific oysters, Crassostrea gigas. Adult oysters were subjected to two diuron concentrations (300 ng L(-1) and 3 microg L(-1)) for 11 weeks. Significantly higher aneuploidy level was observed in diuron-treated oysters compared with the control. Furthermore, the observed impact on aneuploidy persisted to the next generation as offspring exhibited significantly higher aneuploidy levels when their parents had been exposed to diuron. Significant increases in hemocyte parameters (cell mortality, phagocytosis, granulocyte percentage, reactive oxygen species, and lysosome presence) of the adults were also observed after 4 weeks of diuron exposure. The effects observed on oyster aneuploidy level and hemocyte parameters could have serious environmental and practical consequences.

Effects of cadmium on aneuploidy and hemocyte parameters in the Pacific oyster, Crassostrea gigas.

Pacific oysters, Crassostrea gigas, are commonly reared in estuaries where they are exposed to anthropogenic pollution. Much research has been made on the toxicity of cadmium to aquatic organisms because the compound recurrently contaminates their environment. Our study examined the influence of cadmium on aneuploidy level (lowered chromosome number in a percentage of somatic cells) and hemocyte parameters in C. gigas at different stages of life. Adults and juveniles were exposed to two different concentrations of cadmium. The first concentration applied was equivalent to a peak value found in Marennes-Oléron bay (Charente-Maritime, France; 50 ngL(-1)) and the second was 10 times higher (500 ngL(-1)). Exposure to 50 ngL(-1) cadmium caused a significant decrease in the survival time of C. gigas, but exposure to 500 ngL(-1) surprisingly affected the survival time positively. Significant differences in aneuploidy level were observed between the cadmium treatments and the control in adults but not in juveniles or the offspring of the adult groups. The effects of cadmium on hemocyte parameters were analyzed by flow cytometry. Several hemocyte parameters increased significantly after 21 days of cadmium exposure and subsequently decreased. Phenoloxidase-like activity, evaluated by spectrophotometry, varied over the time of the experiment and increased after 66 days of contact with 500 ngL(-1) cadmium. Taken together, cadmium at environmentally relevant concentrations seems to have only moderate effects on aneuploidy and hemocyte parameters.

Endonuclease banding reveals that atrazine-induced aneuploidy resembles spontaneous chromosome loss in Crassostrea gigas.

Aneuploidy has previously been observed in the Pacific oyster, Crassostrea gigas, and shown to be negatively correlated with growth. Moreover, a significant impact of atrazine exposure has been described in C. gigas, and persistence of that effect has been observed between generations. Evidence of differential chromosome loss has been demonstrated in aneuploid karyotypes of C. gigas using the G-banding technique. Pairs 1, 5, 9, and 10 are characterized by the loss of 1 chromosome. As restriction enzyme (RE) digestion chromosome banding allows a better identification of chromosome pairs, we used this technique to identify which chromosomes are affected when aneuploidy is increased by exposure to atrazine. The progeny of oysters contaminated by atrazine were analysed using the restriction enzyme HaeIII. The study of 26 RE-banded aneuploid karyotypes showed that the same chromosome pairs (1, 5, 9, and 10) were affected by the loss of 1 chromosome (61%, 15%, 42%, and 42%, respectively). Further investigation is required to enable a better understanding of aneuploidy in oysters, especially with respect to why some chromosomes are more easily lost than others, and why cells tolerate the loss of these chromosomes.

Impact of atrazine on aneuploidy in pacific oysters, Crassostrea gigas.

Aneuploidy has previously been described and studied in the Pacific oyster, Crassostrea gigas, and has been shown to be negatively correlated with growth. The present study investigated the effect of atrazine on the level of aneuploidy in this species. Crassostrea gigas adults and juveniles were subjected to different concentrations of atrazine representing a peak value found in a polluted environment (46.5 nM) and a value 10 times higher (465 nM). Although atrazine did not show any effect on the oyster mortality, significant differences in aneuploidy level were observed between the different treatments (9% for the control, 16% for 46.5 nM and 20% for 465 nM atrazine). Moreover, the same levels of aneuploidy were observed at adult and juvenile stages. This is the first reported evidence for an environmental effect on aneuploidy in C. gigas. These results will be useful for the oyster aquaculture industry and management of resources. The lowest atrazine level in the current study represents realistic potential exposure, and the results suggest that studies should be made on other aquatic species at risk of exposure to atrazine in the wild. This widely used compound may be an important factor causing damage to genetic material.