Extraction and Purification of Neurotoxin (Anatoxin-a) From Blue-green Alga Pesudoanabaena limnetica and Indicating Its Histopathological Effects on The Brain of Male Laboratory Mice (Mus Musculus L.)

Abstract

The current study included isolating, purifying and cultivation the blue- green alga P.limnetica and recording for the first time in the province of Basra and Iraq and was diagnosed for its ability to produce neurotoxin (Anatoxin-a) .The toxin was extracted and purified from alga and quantify its quantitative and qualitative concentration using ELISA-KITS as it reached 1.179 µg/L for 50mg D.W of (=23.58µ/L for 1gm D.W). The diagnosis of algae as the product of this type of toxins study is the first of its kind on this specific species and the scarcity of species known for the production of neuropathy. The physical properties of the purified toxin were studied using ultraviolet radiation techniques with a single absorbance peak on the wavelength 226 nm and infrared spectrum as most of the active groups in the standard toxin composition. The study also examined for the first time locally and globally the statement of the histopathological effects of the purified toxin on the brain of the laboratory male mice in few concentrations, for a period of 15 days administration, where many of the histopathological effects were observed, which increased with increased concentration and represented decay in the thickness of the gray matter and the karyopicknosis of the nuclei of the glial cell nuclei and shrinkage of neurons at the concentration 0.5 µg/L and  by increasing the concentration to 1 µg/L  there was significant and clear decay in the thickness of the gray area and a decrease in the number of glial cells and the disappearance of most of them with increased shrinkage of neurons. The results showed that there was congestion of the capillaries at the concentration 0.5µg/L and the decay of the The disappearance of the endothelial cells lining the blood vessel and may disappear completely with the expansion and congestion more when the concentration of neurotoxin increased to 1µg/L and the white matter was seen as spongiform at the end of the period of exposure to the toxin and the emergence of cases of hyperpigmentation and edema compared with the control group. The study also showed that physiological effects of anatoxin-a on male laboratory mice represented a significant loss of weight, hair loss and loss of sight within a short period of 7-10 days of exposure to toxin which indicates the seriousness of this type of toxins, So far there were very few concentrations.

Key words: Blue-green algae, P. limnentica, Purification and Extraction, Anatoxin-a, Histopathological effects