Safety study (toxicological evaluation) of one product: marine proteine hydrolysate

CLAUDIA ANESINI; ROBERTO DAVICINO

2008-10-01

2008-12-15

Biológica

Otros campos

PART 1: ANIMAL SAFETY Methodology Acute toxicity of a dried plant extract will be determined in Wistar or Sprague Dowyle rats. For this purpose, initial dosage will be of 100 times the recommended human daily dose. Fourteen animals (7 males and 7 females) will be administered with product by a systemic via (oral - gavage). One group of fourteen animals, males and females will be treated with solvent as control. After administration, the animals will be observed during a period of 30 consecutive days, and the number of deaths will be registered. During this period, general behaviour and signals of acute toxicity principally in central nervous system will be reported, such as: signals of excitability, convulsions, ataxia, hypnosis, CNS depression, and motor coordination, among others. The state of the fur of the animals, and their food and water uptake will be recorded, as well as their weight throughout the whole period of study. The animals weigh will be determined before the product administration (day 0) and after the finalization of observation (day 30). A biochemical control and determination of blood formula and enzymes related to hepatic and renal functionality will be done at the finalization of the study. The laboratory tests that will be performed include: Blood formula (WBC, RBC, platelets, haematocrit, and haemoglobin). Renal functionality will be evaluated through the measure of uric acid, blood urea nitrogen and serum creatinin and hepatic functionality through the determination of SGOT, SGPT and GGT. At the end of the study, the animals will be autopsied; first a pathophysiological observation of intestine, stomach, spleen, liver, test or ovaries, kidneys in comparison with control animals will be done. After this study, the same organs will be maintained in phormaldehyde (10%) for further hystopathological studies. A final report consigning all the results obtained will be written in English, and will be extended officially by the IQUIMEFA-CONICET. PART 2: ADITIONAL STUDIES FOR PRODUCT : Additionally to the safety studies described in PART 1, Product  will be administered once by oral via in BALBc mice (age 2-6 months, 10 females and 10 males). After one month of treatment, the animals will be sacrificed and the intestines will be excised and weighed. Then, the liver will be disaggregated in PBS 1 X and the resulting homogenates will be diluted in sterile saline solution and will be sowed in Petri dishes with an optimum culture medium. At the end of the procedure described, the number of UFC will be recorded. On the other hand, blood samples from animals will be taken and IL-6, IL 10 TNFalfa will be determined using commercial kits.