Review of the Urinary Schistosomiasis Control in Morocco (1960-2018).

The purpose of this study is to describe the epidemiological profile and evolution of urinary schistosomiasis in Morocco, from the first confirmed case in 1960 until disease elimination, and control snails. During this period, 129,526 cases were recorded in Morocco. A majority of cases were reported in Agadir province (25%), Errachidia (18%), and Beni Mellal (13%). Other cases have been reported in the other provinces. Activities within the National Schistosomiasis Control Programme for more than three decades were focused in priori on screening in schools located in high-risk communities, treatment program, surveillance of snails in water bodies, and mollusciciding. Then, the goal of eliminating the transmission of schistosomiasis has been reached in 2004. Sixteen years later, no indigenous cases were detected in Morocco, and only 25 residual cases (resulting from bilharziasis previously treated) are detected, such as in Tata ( 40%), Errachidia (16%), and (12%) in Marrackesh. Similarly, recent national studies conducted on children and the snail reservoir hosts have indicated that no human and molluscs are currently infected with Schistosoma haematobium. Actually, timely investigation and management of imported cases has been implemented to prevent the reintroduction of the disease. The Ministry of Health is planning to implement final confirmatory surveys before requesting WHO to proceed with the formal verification process.

Model of Effective Control of Leishmaniasis in North Central Morocco and Updating the Mapping of the Circulating Species.

Leishmaniasis is among the most important emerging vector-borne protozoal disease in terms of disability adjusted life year (DALY'S). The main objectives of this study are, firstly to evaluate the epidemiological situation of Cutaneous Leishmaniasis (CL) from 2000 to 2015 in Moulay Yacoub Province and Fez Prefecture and secondly to analyze the impact of several factors (geographical and socio-economic factors) on the distribution of CL in these provinces. In addition, this study aims to identify the circulating species of Leishmania in order to justify the cause of the case maintenance and to prevent possible epidemics. The spatio-temporal evolution of CL from 2000 to 2015 in the study area revealed the endemic and epidemic character of the disease with a variation in the number of cases according to provinces, and districts. The sharp decrease of cases observed in the study area could be explained by the several control measures that concern the different actors of leishmaniasis. According to the statistical study, the presence of leishmaniasis is mainly related to population movement and environmental conditions. The results of the clinical identification of cutaneous leishmaniasis in the study area showed a diversity of clinical features with the presence of symptoms miming for L. tropica or L. infantum. This result was confirmed by the molecular study PCR-ITS1-RFLP.

En absence du vaccin et vu la complexité et la diversité des cycles des leishmanioses au Maroc, en plus de la présence des facteurs de risque (urbanisation anarchique, changements climatiques, facteurs socio-économiques…), notre étude s'intègre dans le cadre du Programme national de lutte contre les leishmanioses qui est une des priorités dans les programmes de lutte instaurés par l'OMS. Les objectifs de cette étude sont premièrement d'évaluer la situation épidémiologique depuis le déclenchement de l'épidémie de leishmaniose cutanée (LC) en 2000 jusqu'à 2015 dans la province de Moulay Yacoub et la préfecture de Fès et secondairement d'analyser l'impact de plusieurs facteurs sur la distribution de la LC. Aussi, cette étude vise à identifier les espèces de leishmanies circulantes au niveau de cette région. L'évolution spatio-temporelle de la LC de 2000 à 2015 dans la zone d'étude a révélé le caractère endémoépidémique de la maladie. La nette diminution après le pic de 2001 des cas de la LC peut être liée aux différents moyens de lutte réalisés annuellement et qui ont concerné les différents maillons du cycle. Selon les résultats de l'étude statistique (Ordinary Least Squares Regression [OLSR] et Chi2), la présence de la LC est principalement liée aux mouvements de la population et aux conditions environnementales. Les résultats de l'approche d'identification ont montré une diversité des caractéristiques cliniques avec la présence de symptômes qui indiquent une LC à L. tropica ou à L. infantum. Ce résultat a été confirmé par l'étude moléculaire PCR-ITS1-RFLP.

Poor efficacy of the most commonly used anthelmintics in sport horse nematodes in Morocco in relation to resistance.

Sport and leisure horses in Morocco are treated with several anthelmintics, organophosphates (dichlorvos), benzimidazoles (mostly thiabendazole) or tetrahydropyrimidines (mostly pyrantel pamoate) against nematodes. We studied three horse stables in Rabat, one in Meknes and one in Bouznika. Two of the Rabat and Bouznika stables had introduced a large number of horses from countries (Argentina or Europe) where resistance to benzimidazoles is frequent, whereas the Meknes stud farm remained without foreign introduction. The number of treatments was not very frequent (twice a year in adult horses) but the same anthelmintics were used repeatedly. No resistance to dichlorvos was detected whereas benzimidazole and pyrantel pamoate resistances were detected for the first time in African horses, outside South Africa.

Sodium stibogluconate (Pentostam) potentiates oxidant production in murine visceral leishmaniasis and in human blood.

Sodium stibogluconate (Sbb), a leishmanicidal drug, was studied for its in vivo effect on the formation of reactive oxygen species (ROS), assessed by chemiluminescence (CL) in the whole blood of mice infected with Leishmania infantum. Stimulation of ROS formation induced ex vivo by zymosan particles or the protein kinase C activator phorbol myristate acetate (PMA) was reduced by approximately 25% (P < 0.05) after infection of mice. Treatment of infected mice with Sbb (50 to 400 mg/kg of body weight) enhanced the blood CL induced by zymosan and PMA (47 to 96%, P < 0.01). The drug potentiation effect also occurred in uninfected mice. In vitro treatment of normal human blood with Sbb (1, 10, or 100 microg/ml) for 1 h primed the CL response to PMA (29 to 54%). The priming effect of Sbb was also observed on the production of superoxide by isolated polymorphonuclear leukocytes stimulated either by PMA and zymosan or by the chemoattractants N-formyl-Met-Leu-Phe and platelet-activating factor. These data provide the first evidence of priming of the phagocyte respiratory burst by Sbb. This novel property of Sbb may contribute to the drug's leishmanicidal effect.

Immune responses in vaccinated dogs with autoclaved Leishmania major promastigotes.

A comparative study was undertaken on the immunogen power of autoclaved Leishmania major promastigotes (ALM) vaccines given simultaneously with either BCG or saponin against canine leishmaniasis. The humoral immune response was assessed by ELISA and western blotting. The cellular immune response was evaluated by the lymphocyte transformation test. Dogs vaccinated simultaneously with ALM and saponin showed high antibody titres to crude L. infantum antigens after the first vaccine booster and reacted with several antigens, with molecular weights from 26 to 108 kDa by western blotting. However, the lymphocyte proliferation of these dogs to the crude L. infantum antigen was not significantly different from the control group. In contrast, in dogs vaccinated simultaneously with ALM and BCG, the antibody titres to crude antigen were low. Their sera reacted with the same proteins recognised by sera from dogs vaccinated simultaneously with ALM and saponin by western blotting. However, the 85-kDa protein was only identified by sera taken from dogs vaccinated simultaneously with ALM and BCG. These latter exhibited specific lymphocyte proliferation to the L. infantum antigen. This cell proliferation was observed for approximately 9 months after the first dose of the vaccine. This study indicates that a combination of ALM as the vaccine and BCG as the adjuvant, in the dog model, was successful in inducing a cell immune response, which is implicated in protection of dogs against a Leishmania infection.

Female tick Hyalomma marginatum marginatum salivary glands: preliminary study on protein changes during feeding process and antigens recognized by repeatedly infested cattle.

Proteins extracted from salivary glands of unfed, three days and five days fed adult Hyalomma marginatum marginatum were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). We have noticed changes during the three feeding steps. Some proteins disappeared during feeding process (23, 38, 39, 40 to 50, 95 and 112 kDa), they might be proteins which were converted in other substances and are secreted. Other antigens (13 to 14, 20, 25, 29, 165 and 210 kDa) were synthesized as a result of tick attachment and feeding. They may be related to growth and development or are the ciment which fixed the adult. Also, three Holstein calves were infested five times with 100 pairs of adult ticks of the same species. The five infestations were performed two weeks from the previous infestation. The sera before infestations and after each infestation were used in western-blot analyses to identify antigens from five days salivary gland extracts of the primary infestation of ticks. Three antigens (18.7, 50 and 80 kDa) were revealed weakly after the first and the second infestations by sera samples but not at infestation onward. Others (13.5, 17 to 18.5, 25, 30, 70, 133, 176 and 193 kDa) were revealed only by sera taken after manifestation of resistance (third infestation). A 13.5 kDa antigen was particularly revealed when resistance had appeared and became more evident after the fourth and fifth infestations. The late antigens recognized might be associated with establishment of calves resistance against ticks.

Immunolocalization and characterization of the low molecular weight antigen (4-5 kDa) of Toxoplasma gondii that elicits an early IgM response upon primary infection.

A striking feature of toxoplasmic seroconversion is the prominent and early IgM response to a low molecular weight antigen of 4-5 kDa. Two different monoclonal antibodies directed against the 4-5 kDa antigen have been generated and used to characterize this molecule. Using these monoclonal antibodies, we could demonstrate the surface localization of the low M(r) antigen by immunofluorescence and immuno-electron microscopy assays. By immunoblotting, we observed that one of the monoclonal antibodies was unable to recognize the 4-5 kDa antigen in tachyzoites propagated in cell culture, indicating an epitope variability between Toxoplasma gondii tachyzoites grown in vivo and in vitro. We discuss the implications of this latter finding in the design of diagnostic reagents.

Differential targeting of dense granule proteins in the parasitophorous vacuole of Toxoplasma gondii.

The biosynthesis and fate of 4 different dense granule proteins of Toxoplasma gondii were studied with 3 monoclonal antibodies raised against tachyzoites and 1 polyclonal antibody raised against a recombinant protein. These proteins have the following molecular weights: 27 kDa (GRA 1), 28 kDa (GRA 2), 30 kDa (GRA 3) and 40 kDa (GRA 4). All four proteins were found in dense granules by immunoelectron microscopy; in T. gondii-infected cells, they were found in the vacuolar network but, in addition, GRA 3 was also detected on the parasitophorous vacuole membrane. Therefore, dense granule contents undergo differential targeting when exocytosed in the parasitophorous vacuole. Metabolic labelling and immunoprecipitation showed that GRA 2 and GRA 3 were processed from lower molecular weight precursors, and that GRA 2 and GRA 4 incorporated [3H] glucosamine and are thus likely to be glycosylated.

Surface antigens of Toxoplasma gondii.

Monoclonal antibodies reacting with pellicular antigens of Toxoplasma gondii tachyzoites have been selected among hybridomas produced against this organism by immunofluorescence assay. These antigens have been further characterized by immunofluorescence on living zoites, Western immunoblotting and immunoprecipitation of lactoperoxidase surface radio-iodinated tachyzoite lysates. The simultaneous characterization of 5 different surface antigens (P43, P35, P30, P23, P22) some of which have already been studied individually allowed a better definition of these antigens and the characterization of a yet undescribed surface molecule (P23).

Characterization of a family of rhoptry proteins of Toxoplasma gondii.

A monoclonal antibody (McAb 4A7) raised against a rhoptry enriched subcellular fraction of Toxoplasma gondii tachyzoites reacted in immunofluorescence studies with rod-like organelles located in the anterior part of the organisms and gave specific labeling of rhoptries in immunoelectron microscopy. On immunoblots, two major proteins of 55 and 60 kDa were identified by McAb 4A7. Similar results were obtained both by immunodetection and immunoblotting with tachyzoites, bradyzoites and sporozoites. Pulse chase analysis of [35S]methionine labeled tachyzoites demonstrated that the 55 and 60 kDa rhoptry proteins derived from a 66-68 kDa doublet which was processed approximately 30 min after biosynthesis. Two other monoclonal antibodies (McAb 2F8, McAb 2H3) respectively specific for rhoptry proteins of 55 kDa having a 66 kDa precursor and 60 kDa having a 68 kDa precursor were also obtained; we suggest that they recognize separately the two components of the 55-60 kDa rhoptry protein family of Toxoplasma.

[Influence of the number of inoculated parasites on the in vitro infestation of HeLa cells by the coccidia Besnoitia jellisoni Frenkel]. / Influence du nombre de parasites inoculés sur l'infestation in vitro de cellules HeLa par la coccidie Besnoitia jellisoni Frenkel.

The influence of the number of bradyzoites of the Coccidia Besnoitia inoculated in the culture medium of HeLa cells on their penetration rate was investigated. The intracellular penetration rate increased with the "size of inoculum", then seemed constant when a certain density is reached. Important densities of bradyzoites in the inoculum led to an unequal distribution of intracellular parasites among the cellular sheet. Multiple infective units in a same host cell did not occur randomly. It seems that some cells are more suitable than others to parasite invasion.