Effect of phenyl vinyl sulphone cysteine protease inhibitor on Schistosoma mansoni: in vitro and in vivo experimental studies.

The present work aimed to study the effect of phenyl vinyl sulphone (PVS), a CPI, on different stages of Schistosoma (S.) mansoni in an in vitro culture study and in experimentally infected mice, compared to PZQ. As regards the in vitro study, different concentrations of PVS (1, 2, 4, 6, 8 and 10 µg/ml) and PZQ (1 µg/ml) were assessed by % worm mortality for schistosomula and adults, and hemoglobin degradation by schistosomula. In vivo study included 8 groups of mice. Intraperitoneal PVS, subgroup (a), and oral PZQ, subgroup (b), were assessed at different durations post infection (pi); at 1, 3, 5 and 7 weeks pi (groups I, II, III and IV, respectively). Infection, PVS, PZQ, and normal control groups (groups V-VIII) were included. The anti-schistosomal effects of PVS were assessed by parasitological, histopathological and haematological parameters. In in vitro study, PVS had a schistosomicidal effect in a concentration and time dependent manner, PVS showed 100% schistosomula mortality at day 2 and 92% adult worm mortality at day 5. Furthermore, PVS decreased hemoglobin degradation by schistosomula. In in vivo study, PVS showed a decrease in total worm burden and tissue egg load in intestine and liver with an increase in number of dead ova in intestine of mice. Furthermore, PVS resulted in a decrease in number, size and cellularity of hepatic granulomas and an increase in hemoglobin concentration.PVS was better than PZQ in reducing each of tissue egg count in intestine at 5 and 7 weeks pi, and hepatic granuloma size at 3, 5 and 7 weeks pi. These results suggest that PVS can be a promising chemotherapeutic agent in Schistosoma mansoni infection.

In vivo effect of anti-TNF agent (etanercept) in reactivation of latent toxoplasmosis.

Toxoplasma gondii (T. gondii), an intracellular parasite, establishes a chronic infection by forming cysts preferentially in the brain. TNF-α plays an important role in controlling the infection caused by this protozoan. Thus, the blockade of TNF-α could cause reactivation of latent toxoplasmosis infection as well as increase the risk of acute toxoplasmosis. This study evaluated the effect of etanercept, a TNF-α antagonist in reactivation of latent toxoplasmosis compared to the therapeutic effect of sulfadiazine and pyrimethamine in combination on the progress of the disease. A total of 40 laboratory-bred Swiss albino mice were infected with Me49 strain of T. gondii and divided into four groups: infected control group; treated group with sulfadiazine and pyrimethamine; treated group with etanercept and treated group with both etanercept and sulfadiazine and pyrimethamine. The mean number and size of tissue cysts in brain smears of mice of each group were determined and also, serum levels of TNF-α were assessed in different study groups by an enzyme linked immunosorbent assay. The results showed that the mean TNF-α level was significantly different in the treated groups compared to that in infected control group. The highest level of TNF-α was found in the infected controls. After treatment with etanercept alone or combined with sulfadiazine and pyrimethamine, it was significantly decreased. In this study, reactivation of latent toxoplasmosis was observed by a significant increase in the mean number and sizes of Toxoplasma tissue cysts in brains of mice with established chronic toxoplasmosis after treatment with etanercept alone or combined with conventional treatment compared to both untreated chronically infected controls and infected mice treated with sulfadiazine and pyrimethamine. It was concluded that etanercept, a TNF-α antagonist may play a role in reactivation of latent toxoplasmosis. So, serological screening for toxoplasmosis might offer a valuable aid for patients treated with this drug.

In vitro anti-Trichomonas vaginalis activity of Pistacia lentiscus mastic and Ocimum basilicum essential oil.

Trichomonas vaginalis is a protozoan parasite that causes trichomoniasis; a cosmopolitan sexually transmitted disease. Metronidazole is the drug of choice for T. vaginalis infections. The increase in metronidazole resistant parasites and undesirable side effects of this drug makes the search for an alternative a priority for the management of trichomoniasis. Pistacia lentiscus mastic and Ocimum basilicum oil are known for their antibacterial, antifungal, antiviral and antiprotozoal effects. The present study was carried out to investigate the in vitro effects of P. lentiscus mastic and O. basilicum oil on T. vaginalis trophozoites. The effects of different concentrations of P. lentiscus mastic (15, 10 and 5 mg/ml) and different concentrations of O. basilicum oil (30, 20 and 10 µg/ml) on multiplication of trophozoites at different time points (after 24, 48, 72 and 96 h) were determined, also morphological changes were reported by transmission electron microscopy (TEM). The results showed that both plants caused an inhibition of growth of T. vaginalis trophozoites. The minimal lethal concentration of P. lentiscus mastic was 15 mg/ml after 24 h incubation, 10 mg/ml after 48 h and 5 mg/ml after 96 h. The minimal lethal concentration of O. basilicum oil was 30 µg/ml after 24 h incubation, 20 µg/ml after 48 h and 10 µg/ml after 96 h. TEM study of trophozoites treated by P. lentiscus mastic or by O. basilicum oil showed considerable damage of the membrane system of the trophozoites, and extensive vacuolization of the cytoplasm. These results highly suggest that P. lentiscus mastic and O. basilicum oil may be promising phytotherapeutic agents for trichomoniasis treatment.

A comparative study between excretory/secretory and autoclaved vaccines against RH strain of Toxoplasma gondii in murine models.

Toxoplasma gondii is an obligate intracellular protozoan that has a major importance in public health, in addition to veterinary medicine. Therefore, the development of an effective vaccine for controlling toxoplasmosis is an important goal. Excretory/secretory antigens (ESA), were previously identified as potential vaccine candidates, proved to play important roles in the pathogenesis and immune escape of the parasite. In addition, autoclaved Toxoplasma vaccine (ATV) is a special type of killed vaccine, recently characterized. The aim of the present work was, to compare between excretory/secretory and ATV against RH strain of T. gondii in mice based on; parasitological and histopathological levels. Tachyzoites were harvested from peritoneal exudates of infected mice and were used for challenge infection and vaccine preparation. BCG was used as an adjuvant. Mice were allocated equally into five groups; they were vaccinated intradermally over the sternum. The results of this study showed that the survival time after challenge, extended up to 16 days in ESA vaccinated group and up to 15 days in autoclaved Toxoplasma vaccinated group. ESA vaccinated group exhibited a profound decrease in parasite load following parasite challenge with a higher percentage of reduction in parasite count in all examined organs than the autoclaved Toxoplasma vaccinated group. The histopathological picture of the liver in both immunized groups, revealed marked reduction in the pathological changes observed as compared to controls, especially in ESA vaccinated group. It was concluded that vaccination with ESA showed more promising results versus ATV, as demonstrated by the survival rate of vaccinated mice, tachyzoites count and histopathological examination.

Destructive effect of gamma irradiation on Echinococcus granulosus metacestodes.

Current options for treatment of cystic echinococcosis (CE), including chemotherapeutics and surgical approaches, are not satisfactory in certain cases of resistant cysts in vulnerable or inaccessible organs. Therefore, potential means of therapy are needed. The present work evaluated the effect of gamma irradiation on mestacestode causing CE. Metacestode of Echinococcus granulosus were exposed to 15, 30, and 60 Gy irradiation and examined after 24 h for viability, morphologic, and ultrastuctural alterations by light and electron microscopy. Apoptosis was determined by caspase-3 activity by colorimetric assay and immunohistochemistry. The irradiated metacestodes showed loss of viability, damage of protoscolices, formation of lipid droplets and vacuoles, and separation of the germinal layer. Apoptosis was prominent after irradiation. Our results suggested that gamma irradiation have therapeutic potential in CE. Moreover, understanding the destructive effect of irradiation may help in developing prophylactic measures against CE. Further studies are needed to test the efficacy of ionizing radiation in long-term animal models.

An original simple technique for diagnosis of Microsporidia using glycerol jelly direct smear.

Microsporidia spp. are obligate intracellular parasites which are very minute with sizes ranging from 1 to 10 µm. They have been increasingly recognized as human pathogens in AIDS and immunocompromised patients, mainly associated with life-threatening chronic diarrhea and systemic disease. For accurate identification of Microsporidia, permanent staining techniques are used to enable the examiner to use the ×100 objective which reveals the important details needed for diagnosis. On the other hand, ×10 and ×40 objectives are of no value in detection of such a minute organism. Until now, there is no study that demonstrates a rapid satisfactory technique for routine examination of wet mount by the oil-immersion lens. Glycerol jelly (GJ) reagent was previously studied for its benefit in fixing the cover slide of direct wet mounts instantly enabling the use of oil-immersion lens in examination that magnifies its role as a rapid technique for direct examination. The aim of this research is to identify Microsporidia by wet mounts immediately, using GJ reagent that enables the examiner to use the ×100 objective and to evaluate GJ wet mount as a method of identification. Glycerol jelly reagent was prepared (7 g gelatin dissolved in 50 ml boiling water was added to 10 ml glycerol) and added to fecal wet mounts stained by iodine and methylene blue. Wet mounts were examined using the ×100 objective. Satisfactory results were achieved in spite of the small size of Microsporidia, as both iodine and methylene blue stained the cytological structures; GJ reagent fixed the cover slide, maintained the high translucency of the films, and enabled the examiner to use the ×100 oil-immersion objective. We also compared fecal wet mounts stained by iodine and methylene blue + GJ with a stool sample stained by permanent stain modified Ziehl-Neelsen without GJ, and we found that fecal wet mounts stained by iodine and methylene blue + GJ were more clear. We concluded that glycerol jelly wet mount is an easy, fast, reliable, and cheap technique for identification of Microsporidia in direct smear, using the ×100 oil-immersion objective.