An oral recombinant vaccine in dogs against Echinococcus granulosus, the causative agent of human hydatid disease: a pilot study.

Dogs are the main source of human cystic echinococcosis. An oral vaccine would be an important contribution to control programs in endemic countries. We conducted two parallel experimental trials in Morocco and Tunisia of a new oral vaccine candidate against Echinococcus granulosus in 28 dogs. The vaccine was prepared using two recombinant proteins from adult worms, a tropomyosin (EgTrp) and a fibrillar protein similar to paramyosin (EgA31), cloned and expressed in a live attenuated strain of Salmonella enterica serovar typhimurium.In each country, five dogs were vaccinated with the associated EgA31 and EgTrp; three dogs received only the vector Salmonella; and six dogs were used as different controls. The vaccinated dogs received two oral doses of the vaccine 21 d apart, and were challenged 20 d later with 75,000 living protoscoleces. The controls were challenged under the same conditions. All dogs were sacrificed 26-29 d postchallenge, before the appearance of eggs, for safety reasons.We studied the histological responses to both the vaccine and control at the level of the duodenum, the natural localization of the cestode. Here we show a significant decrease of parasite burden in vaccinated dogs (70% to 80%) and a slower development rate in all remaining worms. The Salmonella vaccine EgA31-EgTrp demonstrated a high efficacy against E. granulosus promoting its potential role in reducing transmission to humans and animals.

Study on the hydatid cyst membrane: permeation of model molecules and interactions with drug-loaded nanoparticles.

The success of the chemotherapeutic treatment of hydatid disease is based upon the drug ability to operate on the germinal layer and on the protoscolices of the hydatid cyst interior at adequate concentrations for sufficient periods. The goal of this study was to evaluate the ability of the drug diffusion through the cyst membrane from sheep hydatid cysts and the increase of drug concentration in the cyst environment. In the first part of this work, the permeation behaviour through the hydatid cyst membrane was studied with five model molecules, having different molecular descriptors (logP, molecular weight, polar surface area ...) onto static Franz glass diffusion cells. A good correlation has been observed between the permeation coefficient and the partition coefficient, log P (r=0.951). In the second part, albendazole-loaded nanoparticles (about 300 nm) prepared by the emulsion solvent evaporation method have shown a sufficient entrapment efficiency (36.4 +/- 6.4%) to raise the apparent solubility of albendazole. The diffusion of drug from the nanoparticles across the hydatid cyst membrane was also improved compare to albendazole suspension. These results have shown the interest of the albendazole-loaded nanoparticles for the treatment of hydatid cysts in the future.

Echinococcus granulosus and other intestinal helminths in semi-stray dogs in Tunisia: infection and re-infection rates.

OBJECTIVE: Assessing the baseline prevalence and the re-infection rate of E. granulosus and other cestodes in dogs is important for a control program based on regular dog dosing treatment with praziquantel mainly to young rural dogs. METHODS: Three hundred and seventy five rural and semi-stray dogs from Jendouba (207 dogs) and Béjà (168 dogs) Departments in Northern-West of Tunisia, were examined to evaluate both the baseline prevalence of intestinal helminths and re-infection rates with Echinococcus granulosus and other cestodes. Parasites were collected in faeces following arecoline hydrobromide purge at the first examination of the dogs and at 4 intervals of reexamination: 2, 4, 8 and 12 months. After each examination, dogs were treated with praziquantel. RESULTS: The global baseline prevalence with Intestinal helminths in dogs was about 35 %. E. granulosus and other helminths were found in 3.5% (E. granulosus); 9.55% (Taenia hydatigena); 6.36% (Taenia pisiformis); 4.77% (Taenia multiceps); 8.59% (Dipylidium caninum), 5.41% (Mesocestoïdes sp.) and Ankylostoma caninum (13.37%). Dogs were re-infected with E. granulosus, T. hydatigena, T. pisiformis and D. caninum, 2 months after the arecoline hydrobromide purge while T. multiceps and Mesocestoïdes sp. infection reappeared 4 months later. CONCLUSION: A control program against Cystic/Echinococcosis, based on a regular treatment of the dog population with praziquantel every 60 days seems necessary. However in practice a six-monthly treatment during ten years would must to decrease the infection pressure of E. granulosus and cease transmission for human.

Molecules targeting the purine salvage pathway in Apicomplexan parasites.

The need of intracellular parasites to retrieve nutrients and fulfill their energy requirements is achieved by manipulating the host's metabolism. With the spread of AIDS, research on purine metabolism has gained in importance with the aim to develop drugs against opportunistic infections. Many studies over the past ten years have yielded contradictory results, but this review tries to clarify these findings by exposing the latest data concerning purine transport and the specific activities of the major enzymes of the purine salvage pathway of Toxoplasma gondii, Plasmodium falciparum and Cryptosporidium parvum.

Antifungal effects of aminosulphoxide and disulphide derivatives.

2-Benzenesulphinyl-(1,4)-naphtoquinone and 14 derivatives were synthesised and were used to evaluate their cytotoxicity against a human myelomonocyte cell line and their antifungal activity against two yeast, i.e. Candida albicans and C. tropicalis and against two filamentous fungi such as Aspergillus niger and Fusarium oxysporum and against one dermatophyte, namely Trichophyton tonsurans. The cytotoxicity and antifungal activities were investigated in comparison with amphotericin B as reference drug. No compound was significantly more toxic than amphotericin B at 0.2 microg ml(-1). The best results of antifungal activity were obtained with GFL 10, GFL 13 and GFL 30 on C. tropicalis, F. oxysporum and T. tonsurans. For C. albicans and A. niger, there was no difference between amphotericin B and the other molecules. The sterol quantitation, the time-kill curves were carried out for these three compounds in order to confirm their action in ergosterol synthesis. Time-kill curves showed a fungistatic activity. For C. tropicalis GFL 10, GFL 13 and GFL 30 increased the growth delay better than amphotericin B, in contrast to F. oxysporum. As for T. tonsurans, GFL10 and GFL13 gave a delay, but the effect of GFL 30 was a bit less marked.

Synthesis and evaluation of antifungal activity of naphthoquinone derivatives.

3-Arylamino-2-phenylsulfinylnaphthoquinones, 2,3-diarylthio-naphthoquinones and 2-phenylsulfinyl-3-arylthio-1,4-dihydronaphtalenes are synthesized and tested against five fungi. The activities of these products were better than amphotericine B against all the strains except for Candida albicans.

[Treatment of hydatid cyst in sheep. Assay of a new scolicide]. / Traitement du kyste hydatique chez la brebis, essai d'un nouveau scolicide.

OBJECTIVE: The main of this study was to show the rapidity of the protoscolicide action of a synthetic compound, dipeptide methyl ester when it is injected under echographic control into hydatid cyst of sheep. MATERIALS AND METHODS: Fourthty sheep with hydatid cysts, repaired at echography and punctionable are treated by dipeptide methyl ester injection at the dose of 110 mM. RESULTS: In vitro tests have allowed to define the efficacy dose of dipeptide methyl ester which is 110 mM. At echography, after injection of the drug, from the first minutes, a detachment of the inner membrane, a diminution of the size of the treated cyst were observed. The cyst content is modified. The sheep autopsy was realized after 4.6.12 and 17 weeks after the injection and showed a size reduction, a treated cyst calcification. CONCLUSION: The dipeptide methyl ester injection into hydatid cyst induces rapidly a morphological alteration, they are calcified. The advantage of this compound is its very rapidity action, this could decrease dissemination risks of hydatid liquid in the organism during operation. Also, this drug permits to reduce the operation time.

Toxoplasma gondii: localization of purine nucleoside phosphorylase activity in vitro and in vivo by electron microscopy.

Purine nucleoside phosphorylase (PNP, EC 2.4.2.1) activity was revealed by enzyme histochemistry in Toxoplasma gondii ME49 strain isolated from murine cerebral cysts and from in vitro cultivation. The activity of the enzyme was revealed by an insoluble electron-opaque precipitate of lead phosphate at the site of the reaction. In bradyzoites and tachyzoites of T. gondii, the enzyme activity could be observed only in the cytoplasm. In bradyzoites, one or two foci of important PNP activity were detected near the nucleus. In tachyzoites, an important PNP activity underlined the plasma membrane. For both bradyzoites and tachyzoites, localization neither in the nucleus nor in cytoplasmic organelles could be detected.

Prevalence of toxoplasma encephalitis in AIDS patients treated with Didanosine hospitalised in a French infectious service.

In a previous work, we have showed in mice infected with an avirulent strain of Toxoplasma gondii and receiving a didanosine treatment, an important decrease of brain cysts. It is why, the purpose of this study was to investigate the effect of didanosine treatment on AIDS patients having developed Toxoplasma encephalitis. 60 patient reports were analyzed: 22 patients (group 1) did not received didanosine in their antiretroviral treatment and 38 (group 2) were treated with didanosine. The results showed that an antiretroviral therapy was prescribed for 93% of patients, 50% of them received only zidovudine and protease inhibitors were prescribed for 37%. The regimens given most frequently were those including zidovudine plus lamivudine or zidovudine plus indinavir. Among the group 1, 18% have had a relapse of Toxoplasma encephalitis. In the group 2, 37% of the patients suffered from one episode of TE while 16% have had two TE after the pause in their didanosine treatment, the maximum occurring between 4 and 24 months after the pause of didanosine. This study showed that didanosine seems to have an effect on cerebral cysts. Also, this work made a synthesis about the different treatment used in AIDS patients and the new molecules yet in development against T. gondii.

Synthesis and antiprotozoal activity of naphthofuranquinones and naphthothiophenequinones containing a fused thiazole ring.

The synthesis of tetracyclic quinones 10a,b, 14a,b, 19a,b and 20a,b is described. The preparations involve regioselective Diels-Alder reactions via trapping the thiazole o-quinodimethane 9 with several benzofuranquinones and benzothiophenequinones. The structure of the regioisomers was assigned through 2D NMR 1H-13C HMBC experiments performed on 10a and 14a. Compounds 10a,b, 14a as well as phenol 1 and the starting quinones 2, 5, 7 and 15 are evaluated against Leishmania sp., Toxoplasma gondii and THP-1 cells. Almost all the tested compounds exhibit significant antiprotozoal activities with lower cytotoxicities than the reference compounds. Among them, quinones 2 and 14a possess the best activities towards L. donovani and T. gondii with the lowest toxicities.

Quinonic derivatives active against Toxoplasma gondii.

Quinonic derivatives were tested against a virulent RH strain of Toxoplasma gondii maintained in cell culture in THP-1, a human myelomonocytic cell line. The derivatives were tested at various doses (0.5-4 microg/ml) and compared with the reference molecules clindamycine, sulfadiazine, pyrimethamine and atovaquone. The percentage of parasite growth inhibition was observed after 72 h of incubation. The tested derivatives are bicyclic, tricyclic or tetracyclic quinones. Eight of these compounds exhibit over 70% inhibition of parasite growth; and two were nearly equipotent to pyrimethamine. These data indicate that the most active compounds against the RH strain of T. gondii are bis-heterocyclic quinones.