Hajiba Fellah

New Clinicoepidemiologic Profile of Cutaneous Leishmaniasis, Morocco

During the past 20 years, cutaneous leishmaniasis has emerged as a major public health threat in Morocco. We describe distribution of Leishmania major and L. tropica in Morocco and a new focus of cutaneous leishmaniasis due to L. infantum. We recommend using molecular techniques to diagnose suspected leishmaniasis cases.

National Serologic Survey of Haematobium Schistosomiasis in Morocco: Evidence for Elimination

The Moroccan Health Ministry launched a Process of Eliminating Schistosomiasis in 1994. During 2005-2009, the epidemiologic status showed a clear interruption of disease transmission at the national level; only a few residual cases were recorded. Our present study is the first systematic serologic survey to evaluate the transmission status in remaining disease-endemic foci. A study population of 2,382 children born after the date of the last autochthonous cases were selected from provinces with histories of high schistosomiasis transmission (Tata, Chtouka Ait Baha, Errachidia, El Kelaa Des Sraghna, and Beni Mellal). To identify the presence of disease, specific antibodies directed against Schistosoma haematobium adult worm microsomal antigens were detected by using an enzyme-linked immunoelectrotransfer blot assay. The results showed an absence of antibodies in all serum samples. Consequently, our findings confirm either a low transmission status or an interruption of schistosomiasis transmission within the last disease endemic foci.

Screening of Antioxidant, Antibacterial and Antileishmanial Activities of Salvia officinalis L. Extracts from Morocco

Aims: The aim of this study was to evaluate in vitro antioxidant and antimicrobial activities of organic extracts from Salvia officinalis L. (Lamiaceae) collected in the province of Ouezzane. Study Design: Evaluation of in vitro antimicrobial and antioxidant activities of extracts and determination phenolic contents. Place and Duration of Study: After plant collection from the Province of Ouezzane, further work was carried out in Parasitology Laboratory of the National Institute of health and Laboratory of Original Research Article Et-Touys et al.; BMRJ, 16(5): 1-10, 2016; Article no.BMRJ.28307 2 Biochemistry-Immunology, Faculty of Science, Mohammed V University of Rabat, Morocco from Novembre 2015 to Mai 2016. Methodology: The antioxidant activity was evaluated using DPPH scavanging assay. The antibacterial activity was tested against three reference strains (Staphylococcus aureus, Escherichia coli and Listeria monocytogenes serovar) by the diffusion method and the minimum inhibitory concentration (MIC) by microtitration assay. The antiparasitic activity was tested against Leishmania major using MTT (3(4.5-dimethylthiazol2yl) -2.5-diphenyltetrazolium bromide) assay. The levels of polyphenols and flavonoids extracts were estimated by colorimetric assay. Results: The methanol extract has shown a significant ability to trap the radical DPPH (IC50=65.655 μg/ml) compared to n-hexane and ethanol extracts. This value is higher than that of ascorbic acid (13.198 μg/ml) and Trolox (22.484 μg/ml) used as standards. The three extracts tests revealed the inhibitory power of three bacterial strains with a significant difference in the diameters of inhibition. The largest area was registered by the hexane and methanol extracts against S. aureus (22±8 mm), while the weakest area was 11±0.22 mm expressed by the ethanol extract against E. coli and the methanol extract against L. monocytogenes. The antileishmanial activity was moderate with a value of cytotoxicity (IC50) above 1 mg / ml. The extracts showed high concentrations of polyphenols and flavonoids, while biological activities were not very high when correlated with these levels. Conclusion: These results will be completed by the determination of the active component and the extracts will be tested on other biological systems namely antifungal and antitumor activities.

Schistosoma haematobium detection in snails by DraI PCR and Sh110/Sm-Sl PCR: further evidence of the interruption of schistosomiasis transmission in Morocco

BackgroundThis is the first study in Morocco to estimate snail infection rates at the last historic transmission sites of schistosomiasis, known to be free from new infection among humans since 2004. Screening of large numbers of snails for infection is one way to confirm that Schistosoma haematobium transmission has stopped and does not resurge.MethodsA total of 2703 Bulinus truncatus snails were collected from 24 snail habitats in five provinces of Morocco: Errachidia, El Kelaa des Sraghna, Tata, Beni Mellal, and Chtouka Ait Baha. All visible snails were collected with a scoop net or by hand. We used waders and gloves as simple precautions. Snails were morphologically identified according to Moroccan Health Ministry guide of schistosomiasis (1982).All snails were analyzed in pools by molecular tool, using primers from the newly identified repeated DNA sequence, termed DraI, in the S. haematobium group. To distinguish S. bovis and S. haematobium, the snails were analyzed by Sh110/Sm-Sl PCR that was specific of S. haematobium.ResultsThe results showed that snails from Errachidia, Chtouka Ait Baha, sector of Agoujgal in Tata and sector of Mbarkiya in El kelaa des Sraghna were negative for DraI PCR; but, snails from remaining snail habitats of El Kelaa des Sraghna, Tata and Beni Mellal were positive. This led to suggest the presence of circulating schistosome species (S. haematobium, S. bovis or others) within these positive snail habitats. Subsequently, confirmation with S. haematobium species specific molecular assay, Sh110/Sm-Sl PCR, showed that none of the collected snails were infected by S. haematobium in all historic endemic areas.ConclusionThe absence of S. haematobium infection in snails supports the argument of S. haematobium transmission interruption in Morocco.

In vitro Antileishmanial Activity of Extracts from Endemic Moroccan Medicinal Plant Salvia verbenaca (L.) Briq. ssp verbenaca Maire (S. clandestina Batt. non L)

Aims: The aim of this study was to evaluate in vitro the antileishmanial activities of organic extracts (methanol, n-hexane and dichloromethane extract) from Salvia clandestina (Lamiaceae) used in Moroccan medicinal plant. Study Design: Evaluation of in vitro antileishmanial activity of extracts and determination phenolic contents. Original Research Article Et-Touys et al.; EJMP, 16(1): 1-8, 2016; Article no.EJMP.27891 2 Place and Duration of Study: After plant collection from the region of Rabat-Morocco, further work was carried out in Parasitology Laboratory of the National Institute of health and Laboratory of Biochemistry-Immunology, Faculty of Science, Mohammed V University of Rabat, Morocco from February 2015 to march 2016. Methodology: The plant was extracted using organic solvents and using Soxhlet. The antileishmanial activity of extracts was tested against three leishmanial strains, Leishmania major, Leishmania tropica and Leishmania infantum in their promatigotes form, using MTT assay. The total phenolic content was assessed by the Folin-Ciocalteau assay and total flavonoid content was assessed by aluminium chloride (AlCl3) colorimetric assay. Results: The MTT based colorimetric assay showed reduced promastigotes viability on the all strains tested. The best growth inhibition was observed with n-hexane and dichloromethane extracts of Salvia clandestina (IC50≤ 155.43 μg/ml) compared to N-methyl glucamine antimoniate (Glucantime®) (IC50>1000μg/ml) used as control, after 72 h of treatment. Phenolic content of S. clandestina extracts ranged between 107.52±3.12 and 74.41±4.96 mg GAE/g extract, and the flavonoid content ranged between 24.64±3.65 and 16.31±3.69 mg QE/g extract. Conclusion: The current investigation reveals that S. clandestina extracts possess activity against three Leishmania species. S. clandestina need further investigation so that the pure bioactive antileishmanial compounds should be isolated with cost effective, promising results and less side effects.

Epidemiological characteristics of visceral leishmaniasis in Morocco (1990–2014): an update

Leishmaniases are parasitic diseases frequent in the Mediterranean Basin. Visceral leishmaniasis (VL) is a notifiable parasitic disease that increased in incidence in Morocco over the past few years and has recently emerged in several new foci, causing a public health problem in Morocco. The aim of this study is to describe the spatio-temporal distribution of VL in Morocco between 1990 and 2014 period in order to highlight important features and trends of VL and its epidemiology and to assess whether the activity of the unit reflects the situation of the disease at the national level and whether it could constitute an indicator of public health relevance. Two thousand four hundred and twenty one cases were reported in Morocco between 1990 and 2014 with an average annual reported incidence rate of 0.4 cases per 100.000 inhabitants. Before 1996 the average annual incidence of VL was 50 cases on average. After this date the number of cases increased and then remained stable with around 100-150 cases per year. Children whose age varies between 1 and 4 years old are the most affected with 1327 (74%) of total cases; nevertheless the adult starts to be affected by the disease. In 2000, 65% of positive cases of VL are concentrated at both northern regions: Taza-Al Hoceima- Taounate with 45% of cases, Tanger- Tetouan mainly represented by Chefchaoun with 20% of cases. The Fez-Boulemane region located in the center recorded 12% of cases. Throughout the years the map VL distribution has been progressively changed and spatial spread of the disease to the center is noted in 2007. 2014 has been marked by an even greater extension of the disease to the center and south of Morocco. Nationally in 2014, 34 of 75 provinces and prefectures are affected compared to 2000, when 22 out of 82 provinces and prefectures were affected. Leishmania infantum was identified the causative agent based on species- specific PCR-Lei70 assay. VL remains a sporadically endemic parasitic disease in Morocco with a progressive extension of its range of distribution. Such a situation would relate to the geographical succession of Phlebotomine sand fly vectors, the difficulty of actions against the canine population reservoirs of L. infantum and unfavorable socio-economic factors.

Comparative evaluation of immunochromatographic dipstick test (ICT) rk39, soluble antigen ELISA and IFAT for the sero-diagnosis of visceral leishmaniasis in Morocco

A rapid, sensitive and specific tool for detection of Leishmania infantum infection in Humans would be highly desirable, because it would allow control interventions in endemic areas of visceral leishmaniasis. This study was carried out at the Reference National Laboratory of Leishmaniasis (RNLL) in National Institute of Hygiene (NIH) Morocco, in order to evaluate the diagnostic potential of immunochromatographic dipstick test (ICT) rk39 in Moroccan suspected VL patients. A total of 49 admitted patients with strong clinical suspicion of VL and 40 healthy controls were investigated for the performance of the ICT rk39. Bone marrow smears were examined for microscopic detection of Leishmania amastigotes obtained from the admitted patients. Only PCR and smear positive cases were considered as gold standard as well as confirmed cases of VL. Out of 49 suspected patients, twenty four (48.9%) were found PCR and smear-positive and twenty three (46.9%) were positive for ICT rk39. Voluntary healthy controls, which included twenty persons from the endemic zone and twenty from non-endemic zone of VL, were found all negative for the strip test. The sensitivity in sera was 75% by ELISA and 87.5% by IFAT, compared with 95.8% for ICT rk39. Specificity was 95.8%, with both tests ELISA and IFAT, and 100% by ICT rk39 respectively. Present study findings again reinforce that the ICT rk39 is a simple, reliable and easy-to-perform non-invasive diagnostic tool for visceral leishmaniasis in the endemic area of Morocco.

Risk factors and prediction analysis of cutaneous leishmaniasis due to Leishmania tropica in Southwestern Morocco

Cutaneous leishmaniasis is currently a serious public health problem in northern Africa, especially in Morocco. The causative parasite is transmitted to a human host through the bite of infected female sandflies of the genus Phlebotomus. The objective of the present study is to characterize the causative organisms and to predict the risk of cutaneous leishmaniasis (CL) cases in six provinces in southwestern Morocco, based on the spatial distribution of cases in relation to environmental factors and other risk factors such as socio-economic status and demographics. A molecular study was carried out using ITS1 PCR-RFLP method of the ribosomal DNA of Leishmania. An epidemiological study on CL cases was reported between 2000 and 2016 in this current investigation in six provinces in southwestern Morocco. Statistical analysis was performed using a linear regression model to identify the impact as well as the interaction between all predictor variables on the distribution of CL in the studied provinces. The forecast Holt-Winters (HW) method was used to describe the trend and seasonality of CL cases. The ITS1-PCR- RFLP analysis revealed the presence of Leishmania tropica in all studied provinces. The spatial distribution of CL cases documented in all studied provinces during the sixteen years showed a heterogeneous pattern and fluctuation trend with an average prevalence of 9.92 per 100,000 inhabitants. In addition, the forecast HW model predicts continued variability of trend and seasonality of CL cases in the upcoming years. This study confirmed the importance of socioeconomic factors, in particular poverty and the vulnerability rate, on distribution and emergence of CL. This study revealed a relationship between increasing risk of CL occurrence due to Leishmania tropica, as well as the distribution and emergence thereof, and socioeconomic factors in the investigated area.

Evaluation d’une grille de supervision des laboratoires des leishmanioses cutanées au Maroc

Introduction The aim of our study was to assess a standardized supervisory grid as a new supervision tool being used in the laboratories of leishmaniasis. Methods We conducted a pilot trial to evaluate the ongoing performances of seven provincial laboratories, in four provinces in Morocco, over a period of two years, between 2006 and 2014. This study detailed the situation in provincial laboratories before and after the implementation of the supervisory grid. A total of twenty-one grids were analyzed. Results In 2006, the results clearly showed a poor performance of laboratories: need for training (41.6%), staff performing skin biopsy (25%), shortage of materials and reagents (65%), non-compliant document and local management (85%). Several corrective actions were conducted by the National Reference Laboratory (LNRL) of Leishmaniasis during the study period. In 2014, the LNRL recorded a net improvement of the performances of the laboratories. The need for training, the quality of the biopsy, the supply of tools and reagents were met and an effective coordination activity was established between the LNRL and the provincial laboratories. Conclusion This trial shows the effectiveness of the grid as a high quality supervisory tool and as a cornerstone of making progress on fight programs against leishmaniases.

Additional file 1: Table S1. of New epidemiological aspects of visceral and cutaneous leishmaniasis in Taza, Morocco

Distribution of leishmaniasis cases in different localities of Taza Province in relation to year (between 2007â 2014) (XLSX 19 kb)