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Original Article

Open Vet J. 2021; 11(1): 112-120

Histological study of the gonads tissues adult stage (male and female) Artemia salina (Linnaeus 1758) and Immunohistochemistry-IHC by Caspase 3 and HSP70 to detect specific apoptosis markers on gonads tissue after exposure to TBTCl

Najla Mohamed Abushaala, Abduofattah Mohamed Elfituri, Syaizwan Zahmir Zulkifli.

Cited by 1 Articles

Background: Several researches have recently been performed on the biological effects of TBTs, including investigations of genitals, in response to exposure to TBTs in marine water for invertebrates.
Aim: The objective of this research to investigate acute effects of TBTCl on gonads in adult stage of Artemia salina by use normal histology and immunohistochemistry (IHC- Caspase3 and HSP70) using for see specific apoptosis markers.
Methods: The samples were analysis about 25 randomly specimens selected from each group of males and females from all different concentrations. The histological method of gonad tissue was used the method for histological preparation Artemia sp protocol. After that, was sectioned the tissues to 5 µm thickness and staining with haematoxylin and eosin. IHC - Avidin-Biotin Complex (ABC-Kit) in this study follow the ABC protocol as suggested by the manufacturer.
Results: finding shown predominant lesions were significant difference (P < 0.05) between the groups. Lesion scores at the different concentration were significantly different from each other concentration showing undifferentiated cells and degenerating. These effects increase in tissue when exposure to the highest concentration of TBTCl compare with control tissue. In testis tissue shows pathological lesions as undifferentiated cells and degenerating testis tissue comparing with control tissue. In female ovary the lesion scores showing undifferentiated cells of the, vitellogenic oocyte with degenerating yolk globules in the ooplasm and degenerating yolk globules and follicle cells enveloping the oocyte increasing these effects in the higher concentration of TBTCl. The representative photomicrograph of testis and ovary section of the control shows no immunoreactivity to caspase 3 and HSP70 antibody. While the representative photomicrograph of testis section showing mild immunoreactivity to caspase3 antibody. And the ovary section showing moderate immunoreactivity to caspase3 following 24hr exposure to TBTCl at 200 ng.L-1. And in HSP70 testis section showing at 100 ng.L-1 mild immunoreactivity to HSP70. ovary section shows at 25ng.L-1 showing moderate immunoreactivity to HSP70, and the positive immunostaining were observed as brownish deposits.
Conclusion: However, there are significant differences between groups in testis and ovary tissue compere with control tissue. In this work IHC is first time used to see effect TBTCl on reproductive organs in A. salina. And caspases3 and HSP70 this study has been proven to possess important function in apoptosis marker and immune system stress in A. salina.

Key words: Keywords: Tributyltin chloride; A. salina; Apoptosis; Immune system stress; Caspase-3; Heat shock protein; Ecotoxicology.

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