Dissected tissues of three shellfish species, the Chinese scallop, Chlamys farreri, Manila clam, Ruditapes philippinarurn, and Razor shell, Solen strictu were evaluated for in vitro transformation of paralytic shellfi...Dissected tissues of three shellfish species, the Chinese scallop, Chlamys farreri, Manila clam, Ruditapes philippinarurn, and Razor shell, Solen strictu were evaluated for in vitro transformation of paralytic shellfish poisoning (PSP) toxins. Tissue homogenates were incubated with extraction from toxic algae Alexandriurn rninutura to determine toxin conversion. The effects of heating and addition of a natural reductant (glutathione) on toxin conversion were also assessed. The toxin profile was investigated through high performance liquid chromatography with fluorescence detection (HPLC-FLD). The evident variations in the toxin content were observed only in Chinese scallop viscera homogenates. The concentration of GTX4 was reduced by 45% (approximately 0.8 μmol/dm^3) and 25% (approximately 1 μmol/dm^3) for GTX1, while GTX2 and GTX3 increased by six times (approximately 1 μmol/dm^3) and 3 times (approximately 0.3μmol/dm^3) respectively. Simultaneously, the total toxicity decreased by 38% during the 48 h incubation period, the final toxicity was 20.4 nmol STXeq/g. Furthermore, heated Chinese scallop viscera homogenates samples were compared with non-heated samples. The concentration of the GTX4 and GTX1 was clearly 28% (approximately 0.53 μmol/dm^3) and 17% (approximately 0.69μmol/dm^3) higher in heated samples, GTX2 and GTX3 were four times (0.66 μmol/dm^3) and two times (0.187 μmol/dm^3) lower respectively. GSH (+) Chinese scallop viscera homogenates samples were compared with GSH (-) samples, the concentration in the GTX4 and GTX1 was 9% (approximately 0.12 μmol/dm^3) and 11% (approximately 0.36 μmol/dm^3) lower respectively, GTX2 and GTX3 was 17% (approximately 0.14 μmol/dm^3) and 19% (approximately 0.006 μmol/dm^3) higher respectively. In contrast,there was a little change in the concentration of PSP toxins of Manila clam and Razor shell tissue ho- mogenates. These observations on three shellfish tissues confirmed that there were species-specific differences in PSP toxins transformation. PSP toxins transformation was more pronounced in viscera tissue than in muscle tissue. PSP toxins was possibly interfered by some carbamoylase enzyme, and the activity in Chinese scallop viscera tissue is more remarkable than in the other two species.展开更多
The current testing for paralytic shellfish poisoning(PSP) in shellfish is based on the mouse bioassay(MBA).To alleviate animal welfare concerns,we evaluated the utility of using sublethal indicators of toxicity as an...The current testing for paralytic shellfish poisoning(PSP) in shellfish is based on the mouse bioassay(MBA).To alleviate animal welfare concerns,we evaluated the utility of using sublethal indicators of toxicity as an alternative to measuring time to death.Live mice were injected with a PSP congener and the changes in neurotransmitter levels were measured 60,90,and 120 min after injection.Acetylcholine(ACh) was the most sensitive marker for PSP toxicity.The changes in neurotransmitter levels were most pronounced in the blood.Thus,measurement of Ach levels in the blood may serve as a sensitive predictor for PSP that would not require sacrifice of the mice.This method was relatively simple,sensitive(less than 1 μg/kg weight,equivalent to 20 ng/mL),low maintenance,and rapid(less than 60 min).展开更多
An inter-laboratory comparison of the AOAC mouse bioassay for paralytic shellfish poisoning (PSP) toxicity in shellfish was carried out among 25 Chinese laboratories to examine the overall performance for PSP testing ...An inter-laboratory comparison of the AOAC mouse bioassay for paralytic shellfish poisoning (PSP) toxicity in shellfish was carried out among 25 Chinese laboratories to examine the overall performance for PSP testing in China, and to analyze the main factors affecting the performance of this method. The toxic scallop Patinopecten yessoensis collected from coast of Bohai Sea, China, was used as a test sample in the comparison study. The results were reported and evaluated using robust statistical methods. The z scores showed that 80%, 8%, and 12% of laboratories reported satisfactory results, unsatisfactory results, and questionable results, respectively. This evaluation demonstrates that the PSP mouse bioassay is an appropriate method for screening and testing PSP toxicity in shellfish. However, it was found that the experience and skill of technicians, as well as the body weight and health status of mice being used significantly affected the accuracy of the method.展开更多
基金The International cooperation programs of the Ministry of Science and Technology of China under contract No.2007DFA30710the Society commonweal programs of the Ministry of Science and Technology of China under contract No.2005DIB2J116
文摘Dissected tissues of three shellfish species, the Chinese scallop, Chlamys farreri, Manila clam, Ruditapes philippinarurn, and Razor shell, Solen strictu were evaluated for in vitro transformation of paralytic shellfish poisoning (PSP) toxins. Tissue homogenates were incubated with extraction from toxic algae Alexandriurn rninutura to determine toxin conversion. The effects of heating and addition of a natural reductant (glutathione) on toxin conversion were also assessed. The toxin profile was investigated through high performance liquid chromatography with fluorescence detection (HPLC-FLD). The evident variations in the toxin content were observed only in Chinese scallop viscera homogenates. The concentration of GTX4 was reduced by 45% (approximately 0.8 μmol/dm^3) and 25% (approximately 1 μmol/dm^3) for GTX1, while GTX2 and GTX3 increased by six times (approximately 1 μmol/dm^3) and 3 times (approximately 0.3μmol/dm^3) respectively. Simultaneously, the total toxicity decreased by 38% during the 48 h incubation period, the final toxicity was 20.4 nmol STXeq/g. Furthermore, heated Chinese scallop viscera homogenates samples were compared with non-heated samples. The concentration of the GTX4 and GTX1 was clearly 28% (approximately 0.53 μmol/dm^3) and 17% (approximately 0.69μmol/dm^3) higher in heated samples, GTX2 and GTX3 were four times (0.66 μmol/dm^3) and two times (0.187 μmol/dm^3) lower respectively. GSH (+) Chinese scallop viscera homogenates samples were compared with GSH (-) samples, the concentration in the GTX4 and GTX1 was 9% (approximately 0.12 μmol/dm^3) and 11% (approximately 0.36 μmol/dm^3) lower respectively, GTX2 and GTX3 was 17% (approximately 0.14 μmol/dm^3) and 19% (approximately 0.006 μmol/dm^3) higher respectively. In contrast,there was a little change in the concentration of PSP toxins of Manila clam and Razor shell tissue ho- mogenates. These observations on three shellfish tissues confirmed that there were species-specific differences in PSP toxins transformation. PSP toxins transformation was more pronounced in viscera tissue than in muscle tissue. PSP toxins was possibly interfered by some carbamoylase enzyme, and the activity in Chinese scallop viscera tissue is more remarkable than in the other two species.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2007AA092001-15)the Key Laboratory of Marine Integrated Monitoring and Applied Technologies of Harmful Algal Blooms,Chinese State Oceanic Administration(No.MATHAB20120101)the Shanghai Municipal Oceanic Bureau(Nos.2011-02,2012-02)
文摘The current testing for paralytic shellfish poisoning(PSP) in shellfish is based on the mouse bioassay(MBA).To alleviate animal welfare concerns,we evaluated the utility of using sublethal indicators of toxicity as an alternative to measuring time to death.Live mice were injected with a PSP congener and the changes in neurotransmitter levels were measured 60,90,and 120 min after injection.Acetylcholine(ACh) was the most sensitive marker for PSP toxicity.The changes in neurotransmitter levels were most pronounced in the blood.Thus,measurement of Ach levels in the blood may serve as a sensitive predictor for PSP that would not require sacrifice of the mice.This method was relatively simple,sensitive(less than 1 μg/kg weight,equivalent to 20 ng/mL),low maintenance,and rapid(less than 60 min).
基金Supported by a thesis research project of General Administration of Quality Supervision, Inspection and Quarantine of China (No. 2010IK168)
文摘An inter-laboratory comparison of the AOAC mouse bioassay for paralytic shellfish poisoning (PSP) toxicity in shellfish was carried out among 25 Chinese laboratories to examine the overall performance for PSP testing in China, and to analyze the main factors affecting the performance of this method. The toxic scallop Patinopecten yessoensis collected from coast of Bohai Sea, China, was used as a test sample in the comparison study. The results were reported and evaluated using robust statistical methods. The z scores showed that 80%, 8%, and 12% of laboratories reported satisfactory results, unsatisfactory results, and questionable results, respectively. This evaluation demonstrates that the PSP mouse bioassay is an appropriate method for screening and testing PSP toxicity in shellfish. However, it was found that the experience and skill of technicians, as well as the body weight and health status of mice being used significantly affected the accuracy of the method.