Moringa oleifera leaf ethanol extract ameliorates lead-induced hepato-nephrotoxicity in rabbits

Objective: To evaluate the effect of Moringa oleifera leaf ethanol extract as an adjunct treatment on lead acetate induced hepato-nephrotoxicity in rabbits. Methods: Thirty-six male New Zealand White rabbits were assigned into two main groups. The first group (14 rabbits) served as normal control. The secondgroup (22 rabbits) was administered orally with lead acetate at a dose of 40 mg/kg/day, 5 days/week for 8 weeks. At the 4th and the 8th week of treatment, 6 animals (3 animals at each period) of the second group were sacrificed while the remaining animals (16 rabbits) were assigned randomly into 2 subgroups (8 rabbits each): treated and non-treated. The first subgroup was orally given 1 mL phosphate-buffered saline for further 4 weeks while the second subgroup was administered orally with Moringa oleifera leaf ethanol extract at a dose of 400 mg/kg/day for the same period. Blood samples were collected to determine hematological and serum biochemical indices. Tissue specimens were collected from the liver and kidney for evaluation of the oxidant/antioxidant markers and for histopathological examinations. Results: Lead acetate exposure decreased the mean body weight gain, hematocrit, hemoglobin, mean corpuscular volume, and lymphocytes count. Moreover, it markedly increased counts of monocytes and platelets, serum enzyme activity, levels of creatinine, total cholesterol, triglycerides, and low-density lipoprotein cholesterol. Malondialdehyde level was markedly increased while the reduced glutathione content was significantly decreased in liver tissue of lead intoxicated-rabbits. Histopathological alterations were also noticed in the liver and kidney of lead intoxicated rabbits. Moringa oleifera leaf ethanol extract significantly improved hematological and serum biochemical parameters and histopathological structure of the liver and kidney. Conclusions: Moringa oleifera leaf ethanol extract ameliorates hemato-biochemical and histopathological alterations caused by lead acetate and improveshepatic and renal functions.

Ultrastructural study on biomphalaria alexandrina haemocytes infected with schistosoma mansoni in Egypt and its correlation with nitric oxide level

Some snails of Biomphalaria alexandrina can resist the infection of Schistosoma mansoni so this study aimed to clearly this mechanism by using light and electron microscopy [EM] and determine the role of Nitric oxide in this mechanism. B. alexandrina snails used in this study were exposed individually to S. mansoni infection according to their response they were classified into susceptible group [shed cercariae] and resistant group [failed to shed cercariae]. Snails not exposed to infection were included in this study as control group. Nitric oxide [NO] level was assayed directly in the soluble fraction of B. alexandrina haemolymph supernatants collected from each group of B. alexandrina snails were subjected to NO assay by the Greiss reaction. The level of NO in haemolymph of infected snails was significantly increased [p<0.001] than both control and non infected snails groups, however, in non infected snails group had significantly [p<0.05] compared to control group. This study when correlated the changes recognized by EM with NO level the pro apoptotic effect of high level of NO on the haemocytes. Characterization and identification of cell shape of haemocytes in both haemolymph and tissue were examined by light and electron microscopy. Examination of B. alexandrina snail's haemocytes revealed three types of different cells classified according to their shape and granular contents. These cells are granulocytes, amoe-bocytes and hyalineocytes. Electron microscope Study also revealed the important role of granulocytes and amoebocytes as defense mechanism against snail infection. NO is considered an important anti parasite molecule; intra-molluscan stages of parasites switch off host NO defense response

Hematological and parasitological effect of cyctein protease inhibitor on murine schistosomiasis

Cysteine protease enzyme [CP] is crucial for parasitic disease propagation, and inhibitors of such proteases are emerging with promising therapeutic uses in the treatment. This study was designed to evaluate the hematological and parasitological efficacy of one of the cysteine protease inhibitors [Sodium nitroprusside] that was administered alone as chemotherapy for murine schistosomiasis manosni in different schistosomal stages [shistosomula and mature worm]. Thirty mice were infected with 80 S. mansoni cercariae/mouse and were divided into 3 groups [10 mice each]. Group I: Infected untreated mice Group II and III. Mice were intraperitoneally treated with 100 mg/kg of cysteine protease inhibitor for 10 consecutive days three and six weeks post infection. Eight weeks post infection all animals were sacrificed and subjected for assessment of cysteine protease protein expression in liver tissue samples using immunoblotting technique [Western blotting], parasitological criteria and EM examination of buffy coat bone marrow and worms. Expression of cysteine protease protein was detected at the expected molecular weight [28 kDa] in 9 of the 10 [90%] infected untreated mice. After treatment protein expression returned negative in the treated groups. A highly significant reduction in worm burden was observed in all groups treated with inhibitor in comparison to infected control group [p<0.001]. However the greatest reduction in the worm burden was observed in treated group 6 weeks post infection in comparison to 3 weeks post infection [P<0.05]. Also, treatment could reduce egg count when given early in infection or significantly decreased egg burden when given late in infection. As regards EM examination sodium nitroprusside treated mice 3 and 6 weeks post the infection revealed degenerated tegument with completely implanted and degenerated spines. In addition 6 weeks post infection treated Schistosoma mansoni male worms showed vaculation and necrosis of spermatocytes. Buffy coat examination of Schistosoma mansoni infected untreated mice showed the inability of the eosinophil to be degranulated, while both treated groups revealed degranulation. In addition the group treated six weeks post infection showed hypodense eosinophils with large number of cytoplasmic vacuoles which represent an activated phenotype. Also in latter group activated lymphocytes were detected with marked dilatation of the endoplasmic reticulum. Bone marrow examination of Schistosoma mansoni - treated mice revealed degranulated eosinophils and eosinophilic myelocytes with activated phenotype in addition to degranulated platelets. Our findings indicate that the cysteine protease secreted by the different stages S. mansoni plays a crucial role in attenuating effector functions of eosinophils as it decreases the eosinophil's responses and inhibit its activation to the parasite resulting in diminished degranulation and reduced generation of superoxide. So, it is preferable to give CPI at any time post infection, with frequent observation of platelets function and different coagulation tests

Ultrastructural and histopathological changes in Giardia lamblia-infected hamsters following administration of two natural compounds and/or metronidazol

The use of combined treatment of metronidazole together with arthemisia and rosemary for controlling intestinal Giardia lamblia infection has been studied. Forty laboratory-bred male hamsters were used in the current experimental study. Each hamster was infected, orally, by 10.000 Giardia lamblia cysts. Animals were divided into the following groups: [1] control infected untreated, [2] infected treated with metronidazole, [3] infected treated with arthemisia and rosemary, [4] infected receiving combined treatment of metronidazole plus arthemisia and rosemary. Each treatment was applied three weeks post-infection. Two weeks later stool analysis was performed and the number of cysts/gm stool was counted, followed by scarification of all animals. The effect of the different drug regimens on the vegetative forms [trophozoites] of the parasite was also studied. There was a highly significant cyst reduction in all treated groups compared to control animals. The highest percentage trophozoite reduction [98.7%] was found in group 4 receiving combined treatment of metronidazole plus arthemisia and rosemary, followed by group 2 [94.8%] and group 3 [62.2%]. By histology, healing of mucosal ulcerations, preserved villi and reduced chronic inflammatory infiltrate in the lamina propria were detected with combined therapy. Ultrastructurual examination of the small intestine in animals of control group showed destructed intestinal cell projections by Giardia lamblia cysts with degeneration of the intestinal submucosa. With combined treatment, complete repair of the intestinal cell projections as well as healing of the mucosa and the submucosa were noticed. Meanwhile, partial healing of destructed intestinal cell projections was observed in group 2 receiving metronidazole alone. It was concluded that the best results were obtained following combined treatment of metronidazole together with arthemisia and rosemary. This might be useful in endemic areas where people tend to develop drug resistance to the commonly used anti-Giardia lamblia preparations

Correlative study between immunodulatory action of propolis on cultured PBMNCs and it's schistosomicidal activity

Propolis, a beehive product widely used in folk medicine as an anti-nflammatory agent, have heen attracting researchers attention to scientifically elucidate us biological properties and therapeutic activities. This study aimed to spot light on the value of propolis as an immune-stimulant and to evaluate the influence on schistosome hematobium infection cure rate. To achieve this goal we estimated the effect of propolis on cultured peripheral blood mononuclear cells activation in-vitro by IL-2 and NO determination. We also evaluated the effect of in-vivo treatment with propolis on Schistosoma hematobium worm and bone marrow by parasitological and ultrastructural studies. Twenty S. haematobium infected golden hamsters were included in the study, subdivided into two groups each of 10 animals Group 1: Infected Control with 300 +/- 10 cercariae of S. haematobium by abdominal skin exposure. Group 2: Animals were treated with propolis three months post the infection. Our in-vitro results revealed that propolis induces a discreet elevation in IL-2 and NO release in PBMNCs cultures supernatant of S. hematobium infected hamsters. Mean level of IL-2 was 16.17 +/- 1.67 pg/ml in the presence of propolis and 3.31 +/- 0.76 in its absence with highly statistically significant difference [p < 0.001]. Regarding NO, Mean level of NO was 7. 76 +/- 1.30 U/ml in the presence of propolis and 2.6 +/- 0.42 in its absence with. highly statistically significant difference [p < 0.001]. Also, propolis caused observed activation and absence of apoptotic changes at the ultrastructural level of cultured PBMNCs revealed. In-viva results, revealed significant reductions in mature worm loads [either male or female], tissue egg loads [either intestinal or hepatic] 21.00 and 19.79% respectively and Percentage reductions of egg developmental stages was 68.07% with statistically significant difference compared with infected control group [P < 0.05]. Ultrastructural study of S. hematobium women revealed implantation and degeneration of the spines within vesiculated tegument and for the bone marrow it revealed evidence of lymphocyte and promonocyts activation in addition to remarkable increase in the number of the activated natural killer cell. Data suggest that propolis acts on host immunity by PBMNCs activation. This information would provide new insights in considering propolis to have a potential therapeutic benefit if used in conjunction with antischistosomal drug in treatment of schistosome infection

Immunolocalization of schistosoma mansoni and schistosoma haematobium antigens reacting with their Egyptian snail vectors

The reaction of the haemolymph and the tissue of infected intermediate hosts, Biomphalaria alexandrina and Bulinus truncatus to Schistosoma mansoni and S. haematobium antigens were investigated using the indirect immunoperoxidase technique. A new technique, Agarose cell block was used in collection of haemolymph which helped in collecting plenty of well formed cells in comparison to the ordinary one using the cytospin. Collected haemolymph and prepared tissues of uninfected and infected B. alexandria and B. truncatus were fixed and then reacted with anti-S. mansoni and anti-S. haematobium IgG polyclonal antibodies. The haemolymph and tissue of infected B. alexandrina and B. truncatus gave a positive peroxidase reaction represented by a brown colour. In haemolymph, the positive peroxidase reaction was detected mainly in the cytoplasm of the amoebocytes. In the tissue, it was detected in epithelial cells lining the tubules, male cells in the lumen of the tubules and in female oogonia cells along the periphery of the tubules. The similarity in the strength and distribution of positive reaction in B. alexandrina and B. truncates was observed as compared to control. Thus, the immunoperoxidase technique proved to be an effective indicator for the schistosome-antigen in the snails

Ultrastructural comparison between the effect of secretory / exceretory products of lung stage and the mechanical S. mansoni schistosomula on each of S. mansoni worm and lymphocyte activation

Ultrastructural observations were made to evaluate the effect of secretory/excretory product [SEP] of 7-days old lung stage S. mansoni schistosomula and the mechanical schistosomula on each of S. mansoni worm, lymphocyte activation and humoral immune response in an attempt to develop a protective vaccine with potent efficiency. Multiple doses [100 micro g followed by two booster doses 50 micro g each] at three weeks intervals were injected intraperitoneally into CFW1 SPE albino mice one week prior to expossure to 100 S. mansoni cercariae, mice were sacrified 6 weeks post infection. Separated worms and lymphocytes in peripheral blood, bone marrow and spleen were prepared for electron microscopic [EM] examination. Serum specific anti-schistosomula SEP IgG was measured. EM examination revealed that immunization with lung stage S. mansoni schistosomula [Group I] induced more damage to the S. mansoni worm represented by extend of the degeneration to subtegumental longitudinal muscle layer, some degeneration in the spermatocyts and vacuolation within the testis of male reproductive organ while degeneration was only whithin the tegument in worm treated with mechanical schistosomula [Group II]. Peripheral blood lymphocytes were activated in both groups while bone marrow lymphocytes were activated only in mice treated with SEP of lung stage S. mansoni schistosomula in which more activation of splenic lymphocytes could be detected in the form of increase number of plasma cells. These results confirmed by the highly significant elevation [p<0.01] in the serum level of anti-SEP IgG in group II. In conclusion, our findings showed that multiple intra-peritoneal administration of low doses of SEP released from 7 days-lung schistosomula induced more damage of the S. mansoni worms, activation of lymphocytes and a significant increase serum level of anti-schistosomula SEP IgG than that of mechanical schistosomula