Search

• Our research investigated the genetic variations among CL, VL, and CanL strains in northern Morocco. • Molecular and genetic analyses uncovered a complex array of genetic variants in the L. infantum and L. tropica species. The number of these variants is 7 and 9 for L. tropica and L. infantum species, respectively. • The Discriminant Analysis of Principal Components (DAPC) revealed significant genetic divergences among L. infantum strains. This suggests the influence of species origin on their genetic makeup. This study investigated nine provinces in northern Morocco and collected 275 skin scraping, 22 bone marrow aspirates, and 89 fine needle aspirations from suspected cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) patients and potentially infected dogs. Molecular analysis using ITS1 RFLP PCR and RT-PCR revealed a higher prevalence of L. infantum (66.18 %; χ2 = 28.804; df = 1; P-value = 8.01e−08) than L. tropica in skin scraping, with L. infantum being the sole causative agent for both VL and canine leishmaniasis. L. infantum was predominantly found in most provinces, while L. tropica was relatively more dominant in Taza Province. Discriminant Analysis of Principal Components (DAPC) revealed distinct clustering between L. tropica and the other three species. However, no small subset of SNPs could clearly differentiate between Infantum_CL, Infantum_VL, and CanL, as they likely share a significant genetic background. The high rate of L. infantum could be attributed to the abundance of sand fly species transmitting VL. In Taza Province, Phlebotomus sergenti, responsible for anthroponotic CL, is the most abundant species. DNA sequencing demonstrated sequence heterogeneity in L. infantum (variants 1-9) and L. tropica (variants 1-7). Phylogenetic analysis showed a distinct separation between L. tropica and L. infantum strains, with an overlap among L. infantum strains isolated from cutaneous, visceral, and canine cases, and dogs serving as the central population for L. infantum. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

In Morocco, Leishmania infantum is the main etiologic agent of human and canine visceral leishmaniasis (VL). This species has been proven to be an opportunistic agent in HIV+ patients and is also responsible of sporadic cutaneous leishmaniasis (CL).This work aims to evaluate the genetic variability of Moroccan L. infantum strains based on PCR-RFLP analysis of the kinetoplastid DNA (kDNA) minicircles. A total of 75 DNA samples extracted from positive Giemsa-stained smears (n = 32) and from L. infantum cultures (n = 43) was studied. The samples have been taken from VL patients infected (n = 7) or not (n = 56) by HIV, patients with CL (n = 2) and finally from infected dogs (n = 10). An hypervariable region of kDNA was amplified using the primers MC1 and MC2; the PCR products were digested separately by a panel of nine restriction enzymes. The presence or absence of restriction fragments was scored in a binary matrix and the SplitsTree4 software was used for the construction of a Neighbor-Net network. Moroccan L. infantum population showed an important level of variability with the identification of 6 genotypes. For each genotype a PCR product was sequenced, confirming the presence of all the expected restriction sites. The predominant profile was the genotype B. A new genotype, named Q was detected for the first time, whereas the four other genotypes (G, K, N and O) were reported sporadically in the Mediterranean basin. The Neighbor-Net network segregates our L. infantum population into 3 clusters: Cluster I includes genotype B, cluster II grouping the genotypes O, Q and G and finally the cluster III contains the genotype N. The kDNA-PCR-RFLP assay is suitable for use directly on biological samples; it reveals an important degree of genetic variability among L. infantum strains even those belonging to the same zymodeme what is of great epidemiological interest. [ABSTRACT FROM AUTHOR]

Visceral and cutaneous leishmaniases are the main endemic vector-born diseases in Morocco. Human visceral leishmaniasis (HVL), by Leishmania infantum , currently presents a significant health problem throughout the country and may constitute factor for death, especially among children with less than 15 years old. In the past, HVL has been basically absent or at least sporadic in Marrakesh-Tensift-Al Haouz region; however it became significant during the last decade. An entomological survey and a retrospective study on L. infantum HVL cases had been carried out to assess the risk of the disease apparition in this region. 7046 sand flies (Diptera: Psychodidae) were collected and studied from twelve localities within Marrakesh-Tensift-Al Haouz region. The result shows the presence of ten sand fly species, 58.76% from the genus Phlebotomus and 41.24% from genus Sergentomyia. A further analysis indicates that Phlebotomus perniciosus, Phlebotomus longicuspis and Phlebotomus ariasi species, incriminated vectors of L. infantum , are dominant (35.56%), so, we describe their spatial (according to altitude and biotopes) and temporal (seasonal activity) distribution in study area. [ABSTRACT FROM AUTHOR]

Abstract: Progressive visceral leishmaniasis (VL) is fatal if not treated; yet, most infections with the causative agents are asymptomatic. We hypothesized that genetic factors contribute to this variable response to infection. The mannose-binding lectin 2 gene (MBL2) is a candidate that merits examination in the context of VL because it enhances infection with intracellular pathogens. Four functional MBL2 polymorphisms at codons 52, 54, 57 and in the promoter at the -221 position (X/Y) are known to be associated with the outcome of several diseases. The aim of the present study was to investigate whether these functional variants were associated with VL in Moroccan children. Here, we genotyped polymorphisms by sequencing and PCR-RFLP in 112 individuals with VL, 97 asymptomatic subjects and 42 healthy individuals who had no evidence of present or past infection. Regression analysis showed no significant association between polymorphisms in exon 1 genotypes and outcome of infection with Leishmania infantum. However, the genotype XY in −221 conferred a protective role against VL in our study population with a significant difference (OR=0.291; CI [0.158–0.538]; p =0.0006). Subjects with YY genotypes in -221 had a higher risk to developing VL. We concluded that MBL2 polymorphism at the -221 promoter region plays a protective role in L. infantum infection. [Copyright &y& Elsevier]

Abstract: Leishmaniases in Morocco are endemic diseases. Three forms of leishmaniasis are reported, visceral leishmaniasis, cutaneous leishmaniasis caused by Leishmania tropica and cutaneous lesions due to Leishmania major. Leishmania infantum, a common parasite inducing visceral leishmaniasis, was observed thereafter in cutaneous lesions. The first case of cutaneous leishmaniasis due to L. tropica was isolated since 1987. But, this parasite was shown to be more polymorphic with almost 8 zymodemes. However, these zymodemes are not all transmitted by Phlebotomus sergenti and not all isolated from human reservoir. Regarding the clinical aspect, cutaneous leishmaniasis with L. tropica is described as a single lesion starting as a nodule at the site of inoculation. A crust develops centrally which may fall away exposing an ulcer which heals gradually. The second cutaneous form is that caused by Leishmania major. It was known in villages located in the southern slopes of the Atlas Mountains. Clinically, the lesion is often severely inflamed and ulcerated and heals in 4–6 months. The epidemiologic cycle of this rural form, include Phlebotomus papatasi as the proven vector and a commensally rodent, Meriones shawi grandis as the reservoir. However, visceral leishmaniasis in Morocco has been known since 1921. It is especially located in the North. The responsible parasite is L. infantum MON 1. Two species of the sand fly are involved in the transmission of this form, P. ariasi and P. perniciosus. In infected man, the clinical signs are non-tender splenomegaly, with or without hepatomegaly, wasting and pallor of mucous membranes. Even though L. infantum MON1 is responsible of the disease, some canine cases were reported to be caused by Leishmania tropica. [Copyright &y& Elsevier]

Abstract: Infantile visceral leishmaniasis (IVL), or Kala Azar, is a parasitic disease caused by the proliferation of a protozoan parasite Leishmania in the reticulo-histiocyte system. It is characterized by its geographical distribution most frequently in the Mediterranean area, India, East Africa and South America. In Morocco, during these last years, we are witnessing a significant recrudescence mainly in young child. The authors performed a retrospective study of 209 cases of visceral leishmaniasis admitted to the service of pediatrics at the University Hospital of Fez (CHU Hassan II) during a six-year period from January 1998 to December 2004. The average age of our patients is 3.7 years and boys are more affected (sex-ratio=1.45) 90% of the cases come from the suburbs of Fez, primarily from Taounate. The average time of consultation is 2.26 months. The main reasons are abdominal distension (68%), fever (94.5%) and the paleness (50%); rarely a hemorrhagic syndrome (8.5%). The clinical examination shows anaemia in 90% of the cases, fever in 92.6% of the cases and splenomegaly in 97.66% of the cases. The diagnosis is confirmed by the myelogram and the serology of leishmaniasis in 91.3% of the cases. Average diagnosis time is eight days. The identification of the parasite is achieved in collaboration with the Pasteur Institute of Casablanca and Leishmania infantum zymodeme-1 is detected in 100% of the cases. The treatment is based on glucantime at the dose of 80 to 100 mg/kg per day during 21 to 30 days and is efficient in 95.57% of the cases. We deplored six deaths which occurred due to malnutrition and probably in association with a macrophage activation syndrome. Infantile visceral leishmaniasis is the most frequent zoonosis in Morocco. Fez and its suburbs are an area of high endemicity. Its eradication must initially be based on improving socioeconomic conditions as well as fighting against malnutrition, which is a risk factor. [Copyright &y& Elsevier]

Abstract: As in the countries edging the Mediterranean basin, Leishmania infantum zymodeme MON-1 is the main causative agent of visceral leishmaniasis in Morocco, where visceral leishmaniasis is most active in the North-Eastern slopes of the Rif mountains. The dog was confirmed to be the main reservoir of L. infantum MON-1, while the reservoir of L. infantum MON-24 causative agent of both infantile visceral leishmaniasis and cutaneous leishmaniasis has not yet been identified. Here we report the first detection of this last zymodeme in a dog in Morocco. The isolated strain was first identified by the use of genotyping markers and confirmed by isoenzyme analysis. Phylogenetic analysis with the use of concatenated sequences from 26 Leishmania donovani complex strains revealed strong geographical correlation with the MON-24 strain from Morocco clustering with other East African strains whereas two other MON-24 strains clustered with L. infantum strains. Interestingly, the two distinct populations of MON-24 identified with the use of genotyping markers cannot be distinguished by multilocus enzyme electrophoresis. [Copyright &y& Elsevier]

• We provide for the first time a model of control program against three major zoonoses. • 67% of total intervention cost was for disease control work with significant economies of scale due to targeting three diseases together. • Our program is efficient on both economic and logistical levels. • This control program could be extended to other countries with the same public health problems. • This study provides effective means of dog population management under the one health approach. The present study is a component of a major European research project, ICONZ (Integrated Control of Neglected Zoonoses). The objective of this component was to implement, and then evaluate an Integrated Control Intervention (ICI) against three dog transmitted zoonoses, namely rabies, visceral leishmaniasis, and cystic echinococcosis. This was undertaken in Sidi Kacem Province (northwestern Morocco) where 22 control and 22 treated douars (villages) were randomly allocated to two control and one evaluation interventions over a period of 13 months. Across the 44 douars, an overall total of 6922 dogs were registered, 4519 were vaccinated against rabies, and 2885 persons attended the complementary health education campaigns; whereas, within the 22 treated douars, 466 dogs received anti-sandfly collars (deltamethrin-impregnated) and 2487 were dewormed (praziquantel). Evaluation of the ICI, revealed that (i) a canine rabies vaccination coverage of 65.0% was achieved, (ii) the use of deltamethrin-impregnated collars for dogs provided highly significant protection (p = 0.01) against leishmaniasis infection of more than 44%, and up to 100% for dogs that had kept the collar until the end of the intervention, (iii) despite a non-significant difference (p >0.05), dog-deworming with praziquantel yielded a reduction in the rate of Echinococcus granulosus infection, and (iv) health education was successful in improving respondents' knowledge; However, the target communities remained unconvinced of the necessity of changing some of their risky behaviours. Lastly, the estimated total cost of the global intervention, including its research components, was US$ 143,050, of which 67% was for disease control work with clear evidence of significant economies of scale due to targeting three diseases together. It appears from this study that the integrated control approach against the three zoonoses was effective on both economic and logistical levels. Image, graphical abstract [ABSTRACT FROM AUTHOR]