Histopathological changes in the skin and gut mucus layers of rainbow trout (Oncorhynchus mykiss) challenged with Ichthyophthirius multifiliis inactivated by gamma rays and formalin

Ichthyophthirius multifiliis, a protozoan parasite, is a significant problem for fish farmers and Aquarium fish worldwide. This study aimed to evaluate the immunization of rainbow trout with gamma-irradiated, formalin inactive, and live theronts of I. multifiliis. In this study, fish were exposed to gamma-irradiated, formalin-inactivated, and live I. multifiliis theronts. Then, the histopathological changes in the mucous layers of the skin and intestines were studied after 7 and 14 days of exposure. Although no significant morphological changes were observed in the skin and intestines of the treated fish, the number of skin goblet cells increased significantly in fish treated with formalin-inactivated, gamma-inactivated, and live trophonts on 7 and 14 days. Compared to the negative control group, an increase in epidermal thickness on the skin was observed in fish challenged with formalin-inactivated, gamma-inactivated, and live trophonts. The numbers of mucous cells/total enterocytes in the intestinal epithelium of fish exposed to gamma-irradiated, formalin-inactivated, and trophonts live were higher than in non-infected fish. Moreover, a significant increase was found in the mucous cell numbers of the pyloric fold in treated fish with gamma-irradiated and formalin inactive trophonts at the first and second weeks. The results showed that the gamma-irradiated trophonts and formalin inactive trophonts could be safe for use in rainbow trout against I. multifiliis

The effects of coadministration of tilorone dihydrochloride and culture supernatants from Lactobacillus reuteri on the mouse hepatoma cell line

Context: Tilorone dihydrochloride is a therapeutic agent with a different mechanism in cancer. The species of Lactobacillus have an important role in cytotoxic effect. Aims: Because of unknown effects of tilorone and culture supernatants from Lactobacillus reuteri on hepatoma, the aim of this study is to evaluate apoptotic, cytotoxic, and therapeutic effects of tilorone on mouse hepatoma cell line with and without culture supernatants from L. reuteri. Materials and Methods: To do so, after cell line culture, cells were divided into different groups such as negative control, treatment with four doses of tilorone, positive control of supernatant (single dose), and combination therapy groups of different doses of tilorone with supernatant (constant doses), for 48 h. All groups were studied with pathologic tests, biochemical study, tetrazolium dye (3-(4, 5- dimethylthiazol -2-yl)-2, 5-diphenyltetrazolium bromide [MTT]) assay, and absolute real-time-polymerase chain reaction (RT-PCR) were done to assess Bax and Bcl-2 genes expression, as molecular studies. Results: MTT assay results revealed that the tilorone tissue culture IC50 (TCIC50) on the Hepa1-6 cell line was 50 μg/ml. RT-PCR analysis showed that tilorone dihydrochloride induced upregulation and downregulation in expression of Bax and Bcl-2, respectively. Simultaneous, antioxidant effect has also seen in a way that prevented necrosis, in biochemical analysis. These results were dose dependent and statistically significant compared to the control group. Conclusions: Based on these results, it appeared that this agent could be a good candidate for further evaluation as effective chemotherapy acting through the induction of apoptosis in hepatoma. The cell death caused through bacterial supernatant was rather necrosis than apoptosis

Developing a Rabbit Model of Neointimal Stenosis and Atherosclerotic Fibrous Plaque Rupture

Background: A precise understanding of the mechanism of human neointimal stenoses and atherosclerotic fibrous plaques, which give rise to thromboses in vital arteries, requires a suitable animal model that would mimic the same characteristics well. We developed a rabbit model of neointimal stenosis and fibrotic plaque rupture in the carotid artery to visualize the lesion progress and to characterize the lesion types according to the American Heart Association classification. Methods: Twenty-eight healthy male New Zealand white rabbits were randomly divided into two groups: The rabbits in group A (n = 14) consumed a standard chow diet, and those in group B (n = 14) were injured via perivascular cold injury using liquid nitrogen at the right common carotid artery before being fed a high cholesterol diet (1.5%) for eight weeks. Plasma lipid evaluation was performed before the sacrificing of the rabbits. At the end of every week, at least 1 rabbit from group B was sacrificed for an analysis of lesion histopathology and calculation of the area ratios of the intima to media. Results: The plasma lipid level in group B was significantly higher than that in group A (p value < 0.05). The histopathological results revealed atherosclerosis characteristics such as endothelial layer destruction, fatty streaks and lipid-containing macrophages (foam cells) formation in the intima and media layers, extracellular lipid collections, smooth muscle cells proliferation and migration, neointima formation, intima thickening and deformation, fibrotic plaque formation, and finally plaque rupture. Statistical analysis revealed a significant increase in the intima-to-media ratio at the end of the eighth week (6.41 ± 0.27, p value < 0.05). Conclusion: We successfully developed a rabbit model of neointimal stenosis and atherosclerotic fibrous connective tissue plaque rupture, which is not only quickly and easily reproducible and inexpensive but also without mortality. The merits of our model render the evaluation of neointimal stenoses and fibrotic plaques and their treatment strategies more feasible in humans