Evaluation of the Effects of Papain on Schistosoma mansoni: Miracidial Infection Capacity, Infection Prevalence, Cercarial Shedding and Molecular Changes in Biomphalaria alexandrina.

PURPOSE: The aim of the present study is to assess the molluscicidal, larvicidal and genotoxicological activities of papain and how it can affect the host-parasite interactions. METHODS: Toxicity of papain on snails by making series of concentrations to calculate LC50, and then study its larvicide effect on the free larval stages of S. mansoni and infection rate of snails. RESULTS: Papain has a molluscicidal activity on adult snails of Biomphalaria alexandrina with a lethal concentration LC50 equals to 43.1 mg/L. In addition, it has activity on miracidia with half Lethal time (LT50) of 16.11 min., and on cercariae with 12.1 min. compared to control ones. The sub lethal concentration LC10 and LC25 (6.9 or 24.1 mg/L, respectively) decreased the survival rate of snails at the first cercarial shedding, the rate of infection, the average total number of cercariae per snail, the shedding period and the life span of snails, while the prepatent period was significantly increased than the control ones. The morphological alterations in cercariae after exposure to papain were occurred where the cercariae lacked motility and some had a dark tail with complete detachment of head and tail. Compared to the control group, the levels of cytochrome oxidase subunit I (COI) and (ND1) genes significantly decreased in snails after exposure to papain. CONCLUSIONS: Papain could be used as a potential molluscicide for elimination of schistosomiasis and decrease its transmission and deterioration of host-parasite interaction.

Evaluation of the prophylactic and therapeutic efficacies of mucus and tissue nucleoproteins extracted from Biomphalaria alexandrina snails on schistosomiasis mansoni.

Schistosomiasis is a neglected tropical disease with considerable morbidity. The lone effective drug, praziquantel (PZQ), is showing emergence of drug resistance hence, searching for new supportive treatment is crucial. This study aimed to evaluate the efficacy of mucus and nucleoproteins (NPs) extracted from Biomphalaria alexandrina (B. alexandrina) snails on miracidia, cercariae and Schistosoma mansoni (S. mansoni) adults in vitro and assess their experimental in vivo effect through parasitological, histopathological, and biochemical parameters. The in vivo study included 90 male Swiss albino mice. Mice were grouped into 9 groups; G1-G5 were infected and treated with; GI: PZQ, GII: mucus, GIII: combined PZQ and mucus, GIV: NPs, GV: combined PZQ and NPs. Control groups; C1: Non infected non treated (negative control), C2: Infected non treated (positive control), C3: Non infected mucus treated and C4: Non infected NPs treated. The in vitro study proved that the mucus had a better lethal effect on cercariae than miracidia, while NPs had better lethal effect on miracidia. The mucus lethal effect on adults surpassed the NPs as 100% and 60%, respectively. The in vivo study proved that the combined NPs or mucus with PZQ added to the effect of individual PZQ resulting in 100% total worm burden (TWB) reduction. As regard oxidative stress markers, the lowest level of nitric oxide (NO) was shown with combined PZQ and NPs. While, the highest glutathione (GSH) level was produced by individual PZQ. The study concluded that mucus and NPs of B. alexandrina had cercaricidal, miracidicidal and anti-schistosomal effect in vitro and that their combination could be considered a contribution to PZQ potentiality in vivo.

Biological, biochemical and genotoxicological alterations of Benzylamine on Biomphalaria alexandrina snails and its Schistosoma mansoni larvicidal potential.

Biomphalaria spp. snails are freshwater gastropods that responsible for Schistosoma mansoni transmission. Schistosomiasis is a chronic illness that occurred in underdeveloped regions with poor sanitation. The aim of the present study is to evaluate the molluscicidal activity of benzylamine against B. alexandrina snails and it larvicidal effects on the free larval stages of S. mansoni. Results showed that benzylamine has molluscicidal activity against adult B. alexandrina snails after 24 h of exposure with median lethal concentration (LC50) 85.7 mg/L. The present results indicated the exposure of B. alexandrina snails to LC10 or LC25 of benzylamine resulted in significant decreases in the survival, fecundity (eggs/snail/week) and reproductive rates, acetylcholinesterase, albumin, protein, uric acid and creatinine concentrations, levels of Testosterone (T) and 17ß Estradiol (E), while alkaline phosphatase levels were significantly increased in comparison with control ones. The present results showed that the sub lethal concentration LC50 (85.7 mg/L) of benzylamine has miracidial and cercaricidal activities, where the Lethal Time (LT50) for miracidiae was 17.08 min while for cercariae was 30.6 min. Also, results showed that were decreased significantly after exposure to sub lethal concentrations compared with control. The present results showed that the expression level of NADH dehydrogenase subunit 1 (ND1) genes and cytochrome oxidase subunit I (COI) in B. alexandrina snails exposed to LC10 or LC25 concentrations benzylamine were significantly decreased compared to the control groups. Therefore, benzylamine could be used as effective molluscicide to control schistosomiasis.

Toxicological effects of Saponin on the free larval stages of Schistosoma mansoni, infection rate, some biochemical and molecular parameters of Biomphalaria alexandrina snails.

Saponins have been used as biopesticides. The objective of the present study is to investigate the toxic effects of Saponin against Biomphalaria alexandrina snails. Results showed that Saponin exhibited a molluscicidal activity against adult B. alexandrina snails at LC50 (70.05 mg/l) and had a larvicidal effect on the free larval stages of Schistosoma mansoni. To evaluate the lethal effects, snails were exposed to either LC10 (51.8 mg/l) or LC25 (60.4 mg/l) concentrations of Saponin. The survival, the infection rates, protein, albumin, and total fat levels were decreased, while glucose levels were increased in exposed snails compared to control snails. Also, these concentrations significantly raised Malondialdehyde (MDA) and Glutathione S Transferase (GST) levels, whereas reduced Superoxide dismutase (SOD) activity and the total antioxidant capacity (TAC) in exposed snails. Furthermore, these concentrations resulted in endocrine disruptions where it caused a significant increase in testosterone (T) level; while a significant decrease in Estradiol (E2) levels were noticed. As for Estrogen (E) level, it was increased after exposure to LC10 Saponin concentration while after exposure to LC25 concentration, it was decreased. Also, LC10 and LC25 concentrations of Saponin caused a genotoxic effect and down-regulation of metabolic cycles in the snails. In conclusion, Saponins caused deleterious effects on the intermediate host of schistosomiasis mansoni. Therefore, B. alexandrina snails could be used as models to screen the toxic effects of Saponins in the aquatic environment and if it was used as a molluscicide, it should be used cautiously and under controlled circumstances.

Gut microbiota modulation as a promising therapy with metformin in rats with non-alcoholic steatohepatitis: Role of LPS/TLR4 and autophagy pathways.

Gut microbiota is a crucial factor in pathogenesis of non-alcoholic steatohepatitis (NASH). Therefore, targeting the gut-liver axis might be a novel therapeutic approach to treat NASH. This study aimed to investigate the therapeutic effects of a probiotic (Lactobacillus reuteri) and metronidazole (MTZ) (an antibiotic against Bacteroidetes) either alone or in combination with metformin (MTF) in experimentally-induced NASH. NASH was induced by feeding rats high fat diet (HFD) for 12 weeks. MTF (150 mg/kg/day) or L. reuteri (2x109 colony forming unit/day) were given orally for 8 weeks; meanwhile, MTZ (15 mg/kg/day, p.o.) was administered for 1 week. Treatment with L. reuteri and MTZ in combination with MTF showed additional benefit compared to MTF alone concerning lipid profile, liver function, oxidative stress, inflammatory and autophagic markers. Furthermore, combined regimen succeeded to modulate acetate: propionate: butyrate ratios as well as Firmicutes and Bacteroidetes fecal contents with improvement of insulin resistance (IR). Yet, the administration of MTF alone failed to normalize Bacteriodetes and acetate contents which could be the reason for its moderate effect. In conclusion, gut microbiota modulation may be an attractive therapeutic avenue against NASH. More attention should be paid to deciphering the crosstalk mechanisms linking gut microbiota to non-alcoholic fatty liver disease (NAFLD) to identify new therapeutic targets for this disease.