Potential toxic effects of titanium oxide (TiO2) nanoparticles on the biological, biochemical, and histological aspects of the land snail Helix aspersa.

Nanotechnology has come a long way in our lives. However, it maintains some negative effects on the environment. This study aims to use the land snail Helix aspersa as a bioindicator. Titanium dioxide nanoparticles (TiO2NPs) had been used at 70 and 140 µg/L for two weeks by the spraying method. The oxidative biomarkers, condition index (CI), DNA damage, hemocyte count, and phagocytic activity were estimated. The toxicity of TiO2NPs was determined (LC50 = 544 µg/L). The exposure to TiO2NPs caused a significant reduction of the activities of superoxide dismutase (SOD) and catalase (CAT) in the digestive gland of Helix aspersa (the activity of CAT was 3.4 ± 0.1 (P = 0.001), SOD was 11 ± 1 (P = 0.0002) at concentration 140 µg/L after two weeks). The activity of glutathione peroxidase (GPX) was (1.13 ± 0.01 µ/mg protein at 140 µg/L compared with controls (5.47 ± 0.01 µ/mg protein). The treatment caused DNA damage in the hemocytes (tail DNA % = 8.66 ± 0.02 and tail moment = 52.99 ± 0 at140 µg/L (P = 0.002)). In the digestive gland, both tail DNA % and tail moment increased (tail moment = 78.38 ± 0.08 compared with control = 2.29 ± 0.09 (P = 0.0001)). The total count of hemocytes significantly decreased after two weeks (the average number was 71 ± 1.5 compared with controls 79 ± 1.1 at 140 µg/L). Furthermore, TiO2NPs caused histological alterations in the digestive gland of Helix aspersa. It can be concluded that the Helix aspersa can be used as environmental pollution bioindicator. A comprehensive evaluation of toxic effects induced by TiO2NPs in vivo assays must be investigated.

Molluscicidal and Larvicidal Potency of N-Heterocylic Analogs against Biomophalaria alexandrina Snails and Schistosoma mansoni Larval Stages.

This work describes the synthesis of quinoline-based N--heterocyclic arenes and their biological evaluation as molluscicides against adult Biomophalaria alexandrina snails as well as larvicides against Schistosoma mansoni larvae (miracidia and cercariae). Molecular docking studies were demonstrated to investigate their affinity for cysteine protease protein as an interesting target for antiparasitics. Compound AEAN showed the best docking results followed by APAN in comparison to the co-crystallized ligand D1R reflected by their binding affinities and RMSD values. The egg production, hatchability of B. alexandrina snails and ultrastructural topography of S. mansoni cercariae using SEM were assessed. Biological evaluations (hatchability and egg-laying capacity) revealed that the quinoline hydrochloride salt CAAQ was the most effective compound against adult B. alexandrina snails, whereas the indolo-quinoline derivative APAN had the most efficiency against miracidia, and the acridinyl derivative AEAA was the most effective against cercariae and caused 100% mortality. CAAQ and AEAA were found to modulate the biological responses of B. alexandrina snails with/without S. mansoni infection and larval stages that will affect S. mansoni infection. AEAA caused deleterious morphological effects on cercariae. CAAQ caused inhibition in the number of eggs/snail/week and reduced reproductive rate to 43.8% in all the experimental groups. CAAQ and AEAA can be recommended as an effective molluscicide of plant origin for the control program of schistosomiasis.

Bioactivity of Trigonella foenum (Fenugreek) oil on immunological and biochemical response of Schistosoma mansoni infected mice.

The effect of fenugreek oil (FO) on some parasitological, immunological, and biochemical parameters in mice infected with Schistosoma mansoni were investigated. Chromatography mass spectrometry (GC/MS) analysis of FO revealed that linoleic acid, (E,E)-4-decadienal, and isopropyl myristate are the major constituents of FO. The results showed that treatment of S. mansoni-infected mice with 0.15 ml of FO daily for 10 successive days exhibited a significant reduction in the number of S. mansoni male worms, and coupled worms as compared to an infected control group (p < 0.05). Regarding total egg counts and oogram patterns, FO effectively reduced the percentage of hepatic and intestinal egg counts, and elevated immature and dead eggs in ratios closely to praziquantel (PZQ) treated mice. Meanwhile, FO significantly elevated the levels of glutathione and co-enzyme Q-10 (COQ-10) up to 0.33±0.02 ng/ml and 0.28±0.02 ng/ml, respectively. However, when accompanied with PZQ, COQ-10 level was closer to that of the normal control group (0.37 ± 0.021 ng/ml). The result also showed that FO significantly reduced levels of lipid per-oxidation (0.165±0.01 ng/ml) and vascular endothelial growth factor (0.25±0.02 pg/ml) as compared to the PZQ-treated group (0.234±0.02 ng/ml and 0.31±0.008 pg/ml, respectively). Moreover, FO recovered normal values of caspase-7, and when accompanied with PZQ, annexin-V was also significantly reduced. However, treatment of S. mansoni-infected mice with PZQ led to a significant increase in the level of annexin-V as compared to S. mansoni-infected mice group (p < 0.05). It can be concluded that FO may have a potential anti-schistosomal, antioxidant and anti-inflammatory activities. Also, it may have a recovering effect on apoptotic parameters toward the normal values.

Protective role of Balanities aegyptiaca fruit aqueous extract in mice infected with Schistosoma mansoni.

The target of this research was to investigate the effect of Balanities aegyptiaca fruit aqueous extract (200 mg/kg BW), alone or in combination with Praziquantel PZQ (300 mg/kg BW) on some biochemical, parasitological, liver histopathology and immunohistochemical parameters in mice infected with Schistosoma mansoni. Results showed that treatment of S. mansoni-infected mice with B. aegyptiaca alone or in combination with PZQ significantly reduced the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) as compared to that of the S. mansoni-infected mice group. Treatment of S. mansoni-infected mice with B. aegyptiaca or PZQ and their combination led to a significant reduction in the activity of malondialdehyde (MDA) as compared with the infected control group. While a significant elevation was observed in the activities of antioxidant enzymes glutathione (GSH), catalase (CAT), superoxide dismutase (SOD) and nitric oxide (NO) compared with the infected. Results revealed that the diameter and number of egg granuloma were significantly condensed after treatment of S. mansoni-infected mice with B. aegyptiaca, PZQ or their combination in hepatic and intestinal tissue. The histopathological alterations observed in the liver of S. mansoni-infected mice were remarkably recovered after B. aegyptiaca treatments. The reduction in angiogenesis was mostly observed in the group receiving the combination of B. aegyptiaca and PZQ. The alterations in vascular endothelial growth factor (VEGF) expression were significantly less in the liver sinusoids. Overall, B. aegyptiaca significantly inhibited the liver and intestinal damage accompanied by schistosomiasis. It demonstrated potent antioxidant and immunoprotective activities. This study advises that B. aegyptiaca can be considered promising for the development of a complementary and/or alternative against schistosomiasis.

Antifungal Activity of Soft Tissue Extract from the Garden Snail Helix aspersa (Gastropoda, Mollusca).

Gastropods comprise approximately 80% of molluscans, of which land snails are used variably as food and traditional medicines due to their high protein content. Moreover, different components from land snails exhibit antimicrobial activities. In this study, we evaluated the antifungal activity of soft tissue extracts from Helix aspersa against Candida albicans, Aspergillus flavus, and Aspergillus brasiliensis by identifying extract components using liquid chromatography-tandem mass spectrometry (LC-MS-MS). Two concentrations of three extracts (methanol, acetone, and acetic acid) showed antifungal activity. Both acetone (1 g/3 mL) and acetic acid extracts (1 g/mL) significantly inhibited C.albicans growth (p = 0.0001, 5.2 ± 0.2 mm and p = 0.02, 69.7 ± 0.6 mm, respectively). A. flavus and A. brasiliensis growth were inhibited by all extracts at 1 g/mL, while inhibition was observed for acetic acid extracts against A. brasiliensis (p = 0.02, 50.3 ± 3.5 mm). The highest growth inhibition was observed for A. flavus using acetic acid and acetone extracts (inhibition zones = 38 ± 1.7 mm and 3.1 ± 0.7 mm, respectively). LC-MS-MS studies on methanol and acetone extracts identified 11-α-acetoxyprogesterone with a parent mass of 372.50800 m/z and 287.43500 m/z for luteolin. Methanol extracts contained hesperidin with a parent mass of 611.25400 m/z, whereas linoleic acid and genistein (parent mass = 280.4 and 271.48900 m/z, respectively) were the main metabolites.

Immune responses, DNA damage and ultrastructural alterations of gills in the marine mussel Lithophaga lithophaga exposed to CuO nanoparticles.

Nanoparticle (NP) pollution is a worldwide problem. Copper oxide nanoparticles (CuO NPs) are one of the most used NPs in a variety of applications, which results in their increased release into the marine environment. In the present work, the marine mussel Lithophaga lithophaga was used as a model organism to evaluate the toxic effects of CuO NPs following 28 days of exposure to sub-lethal concentrations (5 and 20 µg/L). The time points were 1 day of exposure to assess the cell viability, phagocytosis in mussel haemocytes and genotoxicity (DNA damage in gills), 1, 14 and 28 days of exposure to evaluate copper concentrations in water and gills, as well as metallothionein concentration in gills, while gill histology and SEM examination were done after 28 days of exposure. The results indicated that the accumulation of CuO NPs in gills increased with concentration and time. Mussel exposure to CuO NPs increased neutral red uptake. However, the phagocytic abilities decreased in haemocytes with increased concentration. CuO NPs caused DNA damage in the gills even at low concentrations (5 µg/L). CuO NPs caused histopathological alterations in gills, such as brown cell accumulation, necrosis, dwarfism of filaments and ciliary erosion. In conclusion, exposure of the mussel L. lithophaga to CuO NPs led to concentration- and time-dependent responses for all the examined biomarkers. Thus, L. lithophaga may be used as a bioindicator organism in the assessment of CuO NP toxicity.

Oxidative stress and histopathological effect of zinc oxide nanoparticles on the garden snail Helix aspersa.

Nanoparticles have many applications in medicine and biology but they have adverse toxic effects on the biosystem. Therefore, this study aimed to evaluate the toxicity of zinc nanoparticles (ZnO NPs) on the garden snail Helix aspersa. ZnO NPs were used at different concentrations for 7 days. The biomarkers of the oxidative stress and histopathology of the hepatopancreas were estimated. ZnO NPs significantly (p ≤ 0.05) increased catalase (CAT) with time- and concentration-dependent manner. Glutathione S-transferase (GST) activity was significantly (p ≤ 0.05) increased at the concentrations 35 µg/ml and 45 µg/ml after 1 and 3 days of exposure. The present results recorded also a significant elevation in malondialdehyde (MDA) level (time-/concentration-dependent), it was 3.2 ± 0.1, at concentration 45 µg/ml. ZnO NPs induced significant decrease in glutathione (GSH) content (8.7 ± 0.2 at 45 µg/ml) (p ≤ 0.05) at 7 days. Moreover, ZnO NPs induced histopathological alterations in the digestive gland of Helix aspersa. From these results, such biochemical and histopathological alterations in Helix aspersa is a suitable bioindicator of nanoecotoxicological effects.

Anti-neoplastic and immunomodulatory potency of co-treatment based on bovine lactoferrin and/or muramyl dipeptide in tumor-bearing mice.

The current study investigates anti-neoplastic and immunomodulatory activities of co-treatment based on bovine lactoferrin (bLF) and/or muramyl dipeptide (MDP) with or without cisplatin (Cis) in tumor-bearing mice. In the present study, bLF (100 mg/kg; orally) and MDP (0.5 mg/kg; subcutaneously) was administered alone or together. MDP or bLF was co-treated with Cis (1 mg/kg; intraperitoneally) in mice-bearing Ehrlich solid carcinoma. Tumor size, tumor mass proliferation, apoptosis using immunohistochemistry, the alteration in spleen cell proliferation, phenotype using flow cytometry and white blood cells total and differential counts were detected. Treatment with Cis or (bLF and MDP) significantly reduced tumor size, upregulated the pro-apoptotic p53 expression and downregulated the anti-apoptotic Bcl-2 and proliferative marker PCNA expression compared to non-treated tumor-bearing animals. Moreover, co-treatment of MDP and Cis significantly potentiated the reduction of the tumor size, downregulated the Bcl-2 and PCNA expression and upregulated the p53 expression compared to Cis-treated animals. While bLF and Cis co-treatment positively controlled PCNA and p53 expression compared to tumor-bearing animals, it significantly potentiated the reduction of the tumor size and downregulated the Bcl-2 expression compared to Cis-treated animals. Co-treatment of (bLF and MDP), (bLF and Cis) or (MDP and Cis) increased the spleen cell proliferation and altered the immunological profile of the CD3+CD4+, CD3+CD8+, CD3+CD4+CD69+, CD3+CD8+CD69+ and CD11b+Ly6G+ cells to achieve better immune response against tumor. In conclusion, co-treatments based on bLF and/or MDP are promising therapies against cancer, through their potency to control proliferation, enhance apoptosis and improve the immune status against tumor cells.

Toxoplasma gondii: Prevalence of natural infection in pigeons and ducks from middle and upper Egypt using serological, histopathological, and immunohistochemical diagnostic methods.

Toxoplasma gondii is one of the most serious coccidian parasites, which infect human and nearly all warm-blooded animals. Domestic farm birds, which represent a good source of animal protein for humans, are subjected to such infection. Because pigeons and ducks get their feed from the soil, they are susceptible to being infected orally with Toxoplasma oocysts. Consequently, these birds may represent a public health problem for humans. Lack of information on T. gondii infection in pigeons and the insufficient data on its epidemiology in ducks were the main reasons to perform the present study. In the current study, four hundred and fifty-two blood and tissue samples were collected from 310 pigeons and 142 ducks from Asyut, Fayoum, Beni Suef, Minya, and Giza provinces, Egypt. Specific antibodies for Toxoplasma gondii were determined using recombinant TgSAG2t specific surface antigen based ELISA. Histopathological and immunohistochemical (IHC) examinations were also performed. Overall Toxoplasma positivity was 13.55%, 1.61%, and 1.61% in pigeons and 10.56%, 2.11%, and 2.11% in ducks using ELISA, histopathology, and IHC, respectively. Statistically significant difference in the T. gondii prevalence was observed in relation to period of the year. No significant changes were recorded regarding the gender or age. The current study indicated soil contamination and observable Toxoplasma infection in pigeons and ducks, which could represent a major danger for human infection.

Effect of Lactobacillus sporogenes (probiotic) on certain parasitological and molecular aspects in Schistosoma mansoni infected mice.

ABSTRACT: The prospective effects of Lactobacillus sporogenes (probiotics) and/or praziquantel (PZQ) treatment on some parasitological, histological and molecular aspects in Schistosoma mansoni infected mice were studied. The present data recorded that combination between PZQ (300 mg/Kg one dose 7 weeks post infection) and L. sporogenes (12.5 million spore/mice/week for 8 weeks from the first day of infection) reduced worm and ova count. Also, oogram patterns in liver and intestine recorded that treatment with L. sporogenes alone increase number of dead eggs especially in intestine. Histological observations showed significant (P < 0.05) reduction in the mean values of granuloma diameters in liver and intestine in infected mice groups treated with PZQ and/or L. sporogenes. Single strand breaks (comet assay) showed increase in number of damaged and strong damaged lymphocyte cells in mice infected with S. mansoni and infected treated with PZQ while L. sporogenes administration reduced DNA damage. Flow cytometry also confirmed role of L. sporogenes in reducing significantly DNA damage according to determination of cell cycle analysis apoptosis. It can be concluded that administration of L. sporogenes accompanied with PZQ treatment ameliorates the hepatic and intestinal damage caused by S. mansoni infection.

Prevalence of Toxoplasma gondii in Chicken samples from delta of Egypt using ELISA, histopathology and immunohistochemistry.

Estimates of the zoonotic diseases are helpful for monitoring and improving public health. Laboratory-based surveillance provides crucial information for assessing zoonotic disease trends and developments. Toxoplasmosis is considered as a zoonotic disease and has both medical and veterinary importance since it leads to abortion in humans and several animal species. In view of the worldwide importance of T. gondii, this study aimed to estimate the prevalence of T. gondii in chickens from the Delta of Egypt. A total of 304 blood and brain samples were collected from Egyptian chickens from Gharbiya, Qalyoubiya, Minufiya, Beheira, Kafr EL-Shaykh and Dakahlia Provinces. In order to determine the serological and histopathological prevalence of T. gondii, the samples were examined by ELISA, histopathology and immunohistochemistry (IHC). The prevalence of T. gondii was 11.18, 6.91, 6.91 % by ELISA, histopathology and IHC, respectively. Statistically significant differences in the prevalence of T. gondii were observed on the basis of season, sex and habitat. These data provide valuable information regarding the epidemiology of T. gondii infections in Egyptian chickens, which can be employed in developing efficient strategies for disease management and control.

Biomphalaria alexandrina as a bioindicator of metal toxicity.

Heavy metals are common environmental pollutants to the aquatic ecosystems. Several aquatic species have been used as bioindicators and biomonitoring subjects for heavy metals pollution. In the present study, the effects of cadmium (Cd) and manganese (Mn) on the survival, attachment, locomotion, and feeding behaviours of the gastropod snail Biomphalaria alexandrina were determined. The short-term (96 h) LC50 for Cd and Mn were found to be 0.219 and 154.2 mg/l, respectively. Long-term exposures (16-20 days) to ascending concentrations of Cd (0.01-1 mg/l) and Mn (50-500 mg/l) also caused gradual decreases in the survival rate of B. alexandrina in a dose-dependent manner. Attachment, locomotion and feeding behaviours of snails exposed to lethal and sublethal concentrations of Cd and Mn at acute (96 h) and chronic exposure (24 days) intervals, respectively, were also recorded. Compared to controls, a significant decrease (p ≤ 0.05) was recorded in the different behaviours of exposed snails. These changes in behaviour would potentially impact the snail's ability to survive in the wild. Although Cd caused a more severe decline in snail survivorship than Mn, the behavioural effects of Mn were much more severe than Cd when the metals were roughly matched for lethality. In sum, the present study demonstrates B. alexandrina to be a sensitive bioindicator and model organism to assess heavy metals risk factors for severe toxicity in freshwater ecosystems.

Histamine Immunoreactive Elements in the Central and Peripheral Nervous Systems of the Snail, Biomphalaria spp., Intermediate Host for Schistosoma mansoni.

Histamine appears to be an important transmitter throughout the Animal Kingdom. Gastropods, in particular, have been used in numerous studies establishing potential roles for this biogenic amine in the nervous system and showing its involvement in the generation of diverse behaviours. And yet, the distribution of histamine has only previously been described in a small number of molluscan species. The present study examined the localization of histamine-like immunoreactivity in the central and peripheral nervous systems of pulmonate snails of the genus Biomphalaria. This investigation demonstrates immunoreactive cells throughout the buccal, cerebral, pedal, left parietal and visceral ganglia, indicative of diverse regulatory functions in Biomphalaria. Immunoreactivity was also present in statocyst hair cells, supporting a role for histamine in graviception. In the periphery, dense innervation by immunoreactive fibers was observed in the anterior foot, perioral zone, and other regions of the body wall. This study thus shows that histamine is an abundant transmitter in these snails and its distribution suggest involvement in numerous neural circuits. In addition to providing novel subjects for comparative studies of histaminegic neurons in gastropods, Biomphalaria is also the major intermediate host for the digenetic trematode parasite, which causes human schistosomiasis. The study therefore provides a foundation for understanding potential roles for histamine in interactions between the snail hosts and their trematode parasites.

The hepatoprotective activity of blue green algae in Schistosoma mansoni infected mice.

This study aims to evaluate the immunomodulatory effects of a natural product, blue green algae (BGA) (100 mg/kg BW), alone or combined with praziquantel PZQ (250 mg/kg BW) on granulomatous inflammation, liver histopathology, some biochemical and immunological parameters in mice infected with Schistosoma mansoni. Results showed that the diameter and number of egg granuloma were significantly reduced after treatment of S. mansoni-infected mice with BGA, PZQ and their combination. The histopathological alterations observed in the liver of S. mansoni-infected mice were remarkably inhibited after BGA treatments. BGA decreased the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) as well as the level of total protein (TP) while the level of albumin was increased. Treatment of infected mice with BGA, PZQ as well as their combination led to significant elevation in the activities of hepatic antioxidant enzymes glutathione peroxidase (GPX) and glutathione-S-transferase (GST) as compared with control group. Combination of BGA and PZQ resulted in significant reduction in the level of intercellular adhesion molecules-1 (ICAM-1), vascular adhesion molecules-1 (VCAM-1) and tumor necrosis factor-alpha (TNF-α) when compared to those of the S. mansoni-infected group. Overall, BGA significantly inhibited the liver damage accompanied with schistosomiasis, exhibited a potent antioxidant and immunoprotective activities. This study suggests that BGA can be considered as promising for development a complementary and/or alternative medicine against schistosomiasis.