Rhodamine B is one of the industrial food dyes that give cotton candy its bright red color. Antioxidants such as Vitamin C delay or inhibit cell damage primarily through the property of removing free radicals, which alleviate any form of oxidative stress. The aim of the study is to evaluate the effect of daily consumption of rhodamine B with or without vitamin C on some testicular biomarkers, including reproductive organ weight, reproductive hormone levels, and testicular histological examination. Forty-Two adult male rats type albino weighting (150±10g) was divided into 7groups, each group consists of six rats as follows: Group 1: fed on basal diet and used as control (- ve) group. Groups (2,3, and4): fed on basal diet and administrated only rhodamine B at dose 10, 20, and 30mg., respectively. Groups (5,6, and7): fed on basal diet, and administrated rhodamine B at dose 10, 20, and 30mg/200g b.w. and  kiwi juice at 42g/200g BW as a source of  vitamin C, respectively. At the end of experiment(36 day),  Blood samples were taken after the experiment to assess serum testosterone (TH), luteinizing hormones (LH), follicle stimulating hormone (FSH),  serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), oxidative enzyme activity such as superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), lipid profile, blood parameters ,histopathological and biological investigation were also measured. Results indicated that the addition of vit. C improved enzymes CAT and SOD and decreased MDA, also, reduced the harmful effect of rhodamine B on liver enzymes. The intake of vit. C also improved hyperlipidemia, blood parameters, as well as a significant increase in weight, feed intake and food efficiency and improved sex hormones of rats received rhodamine dye and vit. C, which led to a decrease in the negative effect of dye levels.