okadaic-acid and Body-Weight

Okadaic acid (OA) is one of the most prevalent marine phycotoxin with complex toxicity, which can lead to toxic symptoms such as diarrhea, vomiting, nausea, abdominal pain, and gastrointestinal discomfort. Studies have shown that the main affected tissue of OA is digestive tract. However, its toxic mechanism is not yet fully understood. In this study, we investigated the changes that occurred in the epithelial microenvironment following OA exposure, including the epithelial barrier and gut bacteria. We found that impaired epithelial cell junctions, mucus layer destruction, cytoskeletal remodeling, and increased bacterial invasion occurred in colon of rats after OA exposure. At the same time, the gut bacteria decreased in the abundance of beneficial bacteria and increased in the abundance of pathogenic bacteria, and there was a significant negative correlation between the abundance of pathogenic bacteria represented by Escherichia/Shigella and animal body weight. Metagenomic analysis inferred that Escherichia coli and Shigella spp. in Escherichia/Shigella may be involved in the process of cytoskeletal remodeling and mucosal layer damage caused by OA. Although more evidence is needed, our results suggest that opportunistic pathogens may be involved in the complex toxicity of OA during OA-induced epithelial barrier damage.

Topics: Animals; Body Weight; Coleoptera; Colon; Escherichia coli; Okadaic Acid; Rats

Okadaic acid (OA) and the structurally related compounds dinophysistoxin-1 (DTX1) and dinophysistoxin-2 (DTX2) are marine phycotoxins that cause diarrheic shellfish poisoning (DSP) in humans due to ingestion of contaminated shellfish. In order to guarantee consumer protection, the regulatory authorities have defined the maximum level of DSP toxins as 160 µg OA equivalent kg-1 shellfish meat. For risk assessment and overall toxicity determination, knowledge of the relative toxicities of each analogue is required. In absence of enough information from human intoxications, oral toxicity in mice is the most reliable data for establishing Toxicity Equivalence Factors (TEFs).. Toxins were administered to mice by gavage, after that the symptomatology and mice mortality was registered over a period of 24 h. Organ damage data were collected at necropsy and transmission electron microscopy (TEM) was used for ultrastructural studies. Toxins in urine, feces and blood were analyzed by HPLC-MS/MS. The evaluation of in vitro potencies of OA, DTX1 and DTX2 was performed by the protein phosphatase 2A (PP2A) inhibition assay.. Mice that received DSP toxins by gavage showed diarrhea as the main symptom. Those toxins caused similar gastrointestinal alterations as well as intestine ultrastructural changes. However, DSP toxins did not modify tight junctions to trigger diarrhea. They had different toxicokinetics and toxic potency. The lethal dose 50 (LD50) was 487 µg kg-1 bw for DTX1, 760 µg kg-1 bw for OA and 2262 µg kg-1 bw for DTX2. Therefore, the oral TEF values are: OA = 1, DTX1 = 1.5 and DTX2 = 0.3.. This is the first comparative study of DSP toxins performed with accurate well-characterized standards and based on acute toxicity data. Results confirmed that DTX1 is more toxic than OA by oral route while DTX2 is less toxic. Hence, the current TEFs based on intraperitoneal toxicity should be modified. Also, the generally accepted toxic mode of action of this group of toxins needs to be reevaluated.

Topics: Administration, Oral; Animals; Body Weight; Chromatography, High Pressure Liquid; Female; Heart; Intestine, Small; Liver; Mice; Myocardium; Okadaic Acid; Protein Phosphatase 2; Pyrans; Stomach; Tandem Mass Spectrometry; Toxicity Tests

Topics: Administration, Oral; Animals; Body Weight; Chromatography, Liquid; Eating; Feces; Female; Intestines; Lethal Dose 50; Liver; Marine Toxins; Mice; Okadaic Acid; Pyrans; Tandem Mass Spectrometry; Toxicity Tests

The mouse bioassay (MBA) for diarrhetic shellfish poisoning (DSP) toxins has been widely used in many countries of the world. However, different body weight ranges of mice are designated to be used in the Japanese official method and European Union procedure. In this study we investigated whether and to what extent the body weights of the mice affect the susceptibility to DSP toxins. A lethal dose of okadaic acid, one of the representative DSP toxins, was injected intraperitoneally into mice of five different body weight range groups, from 14 to 24 g. The mice were observed until 24 h after injection. The lethality was 100% in the 14-15 and 16-17 g groups, 80% in the 19-20 g group, 50% in the 21-22 g group, and 40% in the 23-24 g group, with significant differences. Survival analysis indicated a relationship between body weights of mice and susceptibility to okadaic acid. These results would be quite useful not only for the MBA, but also to improve understanding of the biological responses to DSP toxins.

Topics: Animals; Biological Assay; Body Weight; Male; Mice; Mice, Inbred ICR; Okadaic Acid; Shellfish Poisoning; Specific Pathogen-Free Organisms

To challenge the theory of tissue specificity of tumor promoters, the biochemical and tumor promoting effects of okadaic acid (OA), a potent tumor promoter on mouse skin, were studied in the mucosa of rat glandular stomach. OA strongly inhibited protein phosphatases 1 and 2A, and increased 4-fold the phosphorylation of elongation factor 2 in vitro in the mucosa. Intubation of 10 micrograms (12.4 nmol) OA induced ornithine decarboxylase in the mucosa. Tumor promotion of OA was studied in the glandular stomach initiated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in a two-stage carcinogenesis experiment. OA in drinking water, 10 micrograms (12.4 nmol) per rat per day from weeks 9-55 of the experiment, and 20 micrograms (24.8 nmol) from weeks 56-72, significantly enhanced development of the neoplastic changes in the glandular stomach (P < 0.05). The neoplastic changes included adenomatous hyperplasias and adenocarcinomas, both of which correspond to papillomas and carcinomas in a two-stage mouse skin carcinogenesis experiment. The percentages of neoplastic change-bearing rats of the groups treated with MNNG plus OA, MNNG alone or OA alone were 75.0, 46.4 and 0% respectively. OA enhanced tumorigenesis in the MNNG-initiated glandular stomach of rats through the same mechanisms of action as in mouse skin. The OA pathway mediated through inhibition of protein phosphatases 1 and 2A is applicable to various organs as a general mechanism of tumor promotion.

Topics: Animals; Body Weight; Carcinogens; Enzyme Induction; Ethers, Cyclic; Female; Gastric Mucosa; Male; Methylnitronitrosoguanidine; Okadaic Acid; Organ Specificity; Ornithine Decarboxylase; Phosphoprotein Phosphatases; Rats; Rats, Inbred F344; Rats, Sprague-Dawley; Stomach; Stomach Neoplasms

The standard mouse bioassay, used to assess 'diarrhoetic shellfish poison' (DSP), is based on intraperitoneal administration of toxic mussel extracts, and monitoring of survival time within a 24-hr period. Toxic effects on mice were examined for extracts of mussel samples from two different regions of south Norway known to possess toxins of specific properties. Both samples revealed an exponential pattern in the dose-response relationship. Whereas the time lag from injection to death was linearly dependent on mouse weight, the effect of weight also increased with decreased sample toxicity. When tested with doses adjusted for weight, a marked individual variation was found within all size groups of mice. The results imply that, regarding prohibition limits for distribution and sale of mussels, a certain degree of variation with regard to time should be accepted in the testing of parallel samples. On the basis of the results, a revised method for the determination of toxicity by mouse bioassay is proposed for DSP testing. The method is based on administration to two mice of size-adjusted doses of extracts, followed by a 4-hr surveillance period and a 1-hr upper limit of acceptable time variation between parallel samples. The method shows advantages regarding savings of time and money, in precision in determination of toxicity level, as well as curtailed exposure to toxin and reduced suffering of laboratory animals.

Topics: Animals; Biological Assay; Bivalvia; Body Weight; Diarrhea; Dose-Response Relationship, Drug; Ethers, Cyclic; Marine Toxins; Mice; Okadaic Acid; Pyrans