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• 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]

This article discusses a case of cutaneous erysipeloid leishmaniasis caused by Leishmania infantum in Morocco. The patient, a 46-year-old farmer, initially presented with swelling of the left lower eyelid and was treated for chronic dacryocystitis. However, the dermatological examination revealed an erysipeloid erythematous placard on the face. The diagnosis of cutaneous leishmaniasis was confirmed, and the patient was treated with antimony salts and metronidazole, showing partial improvement. The article highlights the evolving epidemiological situation of leishmaniasis in Morocco and the atypical erysipeloid form of the disease. The authors emphasize the importance of considering cutaneous leishmaniasis in the diagnosis of unusual facial lesions resembling erysipelas. [Extracted from the article]

Zoonotic visceral leishmaniasis is a common vector‐borne systemic disease caused by Leishmania infantum (Kinetoplastida: Trypanosomatidae). In Morocco the situation is complex: many sandfly species have been collected in areas in which the disease is endemic, but only Phlebotomus ariasi, Phlebotomus perniciosus and Phlebotomus longicuspis (Diptera: Psychodidae) have been confirmed to have vectorial roles. The objective of the present study was to ascertain the potential distribution of L. infantum and its vectors in Morocco, using ecological niche modelling. Vector records were obtained from field collections of the Laboratory team and from previously published entomological observations. Epidemiological data for L. infantum modelling were obtained from Moroccan Ministry of Health reports. The jackknife test indicated that the bioclimatic variables with the greatest influence on model development for all species were annual precipitation and precipitation in the driest quarter of the year. MaxEnt model representations for sandfly species that act as vectors of L. infantum showed the widespread geographic distribution of these species in Morocco, specifically in northern and central Morocco, where foci of visceral leishmaniasis are found. The ecological niche modelling points out areas in which the probability of occurrence of these species is higher. This information should be considered as a starting point for further research to fully elucidate the ecology and epidemiology of these species, as well as of the pathogens they transmit. [ABSTRACT FROM AUTHOR]

Background. Human leishmaniasis, both visceral and cutaneous, has been reported in Morocco for centuries and constitutes a serious public health problem. However, the evolution of this pathology depends on several factors such as ecological, socioeconomic, and climatic conditions. The risk study of the affected foci is of great value for the control and surveillance of this endemic disease, especially in the provinces where Leishmania infantum predominates. Methods. This study concerned nine provinces located in the extreme and central north of Morocco (Taounate, Taza, Chefchaouen, Al Hoceima, Larache, Tétouane, Tanger-Assilah, M'diq-Fnideq, and Fahs-Anjra Provinces). In this work, leishmaniasis cases (VL and CL) were subjected to an epidemiological study which was performed using a linear regression model to identify the impact as well as the interaction between all predictor variables on the distribution of leishmaniasis in this region. Results. During the period 1997–2018, a total of 6 128 cases of VL and CL were recorded in the study area. Our results showed that among demographic factors studied, urbanization showed significance for both cutaneous and visceral forms (P < 0.05). Regarding the environmental factors, the humidity and the altitude were significant for both CL and VL (P < 0.05), while the temperature and the normalized difference vegetation index (NDVI) showed a significance only for VL. Moreover, trends in season of occurrence revealed that wet season (October to April) had a higher incidence of leishmaniasis compared to the dry season (May to September) specifically for CL. As for socioeconomic factors, poverty was the only factor that influences the spread of VL. Finally, the distance from endemic foci showed significance for both VL and LC (P < 0.05). Conclusion. Our study revealed that the risk factor associated with cutaneous and visceral leishmaniasis in northern Morocco could help in the establishment of a prediction program. [ABSTRACT FROM AUTHOR]

Increasing cases of leishmaniasis disease have been reported during recent years in Sefrou Province, Central North of Morocco. This study presents the epidemiological profile of the provincial population, aims at analyzing the epidemiological profile, and in particular, spatiotemporal follow-up of all cases of leishmaniasis. It is a retrospective analysis of leishmaniasis cases recorded between 2007 and 2010. The data were analyzed by SPSS software (version 20). Over a four-year period, from 2007 to 2010, there were 62 cases of leishmaniasis, 93.12% of cases of cutaneous leishmaniasis and 6.87% of visceral leishmaniasis. The case number of leishmaniasis in the Province of Sefrou varies between 0.165% and 0.0018%. For each type of leishmaniasis, the female sex was the most affected compared to the male sex. This difference cannot be considered statistically significant (χ2 = 0.083, p value = 0.77). For cutaneous leishmaniasis, all age groups were affected with a large percentage: patients aged 0–9 years with 63.11% followed by the age group [10–19] with 24.18%. Visceral leishmaniasis mainly has affected the infant population [0–9] with 83.33%. We have not observed any association between the age classes and the leishmaniasis type (χ2 = 6.20, p value = 0.4). From a spatial point of view, the majority of cases of leishmaniasis was reported in El Menzel region (67 cases) followed by Sefrou (64 cases) and Tazouta (38 cases). There is a statistically significant relationship between the type of leishmaniasis and the studied regions (χ2 = 52; p value <0.001). The study of the epidemiological profile of leishmaniasis cases may be useful in enlightening health authorities to develop screening, treatment, and control strategies to reduce the incidence rate of the disease. Other research studies can be conducted to the dynamics of the vectors of sandflies and their ecology. [ABSTRACT FROM AUTHOR]

Background: Zoonotic visceral leishmaniasis (VL) is endemic in the Mediterranean basin. However, large-scale comparative analyses of the commercial kits for the serological diagnosis of this neglected disease are lacking. This study compared the performances of four enzyme-linked immunosorbent assays (ELISA) and two immunochromatographic tests (ICT) as screening tests for the serodiagnosis of human VL in the Mediterranean region. Methodology/Principal findings: Serum samples from 319 patients living in France, Tunisia or Morocco were tested using two ICT (IT LEISH and TruQuick LEISH IgG/IgM Meridian) and four ELISA reagents (NovaLisa Leishmania infantum IgG, Bordier Leishmania infantum, Ridascreen Leishmania IgG, and Vircell Leishmania). The population with proven VL (n = 181) included 65 immunocompromised patients. Significantly higher percentages of false-negative results were obtained with all assays in immunocompromised patients, compared with the immunocompetent population. In the whole population, sensitivity and specificity ranged from 80.7% to 93.9% and from 95.7% to 100%, respectively. The maximum accuracy was observed with the Bordier and Vircell ELISA kits (96.2%), and the lowest accuracy with Ridascreen reagent (88.7%). New thresholds of positivity are proposed for the Bordier, Vircell and NovaLisa ELISA kits to achieve 95% sensitivity with the highest possible specificity. Western blot (WB), used as a confirmation method, showed 100% sensitivity and identified 10.1% of asymptomatic carriers among the control population from the South of France. Conclusions/Significance: This is the first study that compared commercially available kits for VL serodiagnosis in the endemic region of the Mediterranean basin. It provides specific information about the tests' performance to help clinicians and biologists to select the right assay for VL screening. Author summary: Human visceral leishmaniasis (VL) is a neglected disease endemic in the Mediterranean region, caused by Leishmania infantum. Accurate VL diagnosis is critical for rapid and correct patient management. Various serological tests are used, including commercial kits that require qualifications with specific information about their performance. In this study, we compared the performance of six commercial kits, two immunochromatographic tests (ICT; IT LEISH and TruQuick LEISH IgG/IgM Meridian) and four enzyme-linked immunosorbent assays (ELISA; NovaLisa Leishmania infantum IgG, Bordier Leishmania infantum, Ridascreen Leishmania IgG, and Vircell Leishmania), available in France and used as screening tests. The Bordier and Vircell ELISA reagents displayed the highest analytic performance compared with all the other kits. However, no test displayed 95% sensitivity at the thresholds recommended by the manufacturers. To reach this sensitivity value with the highest possible specificity, new thresholds are proposed for the Bordier, Vircell and NovaLisa ELISA reagents. Positive and negative results obtained with the six screening tests were confirmed by Western blot (100% sensitivity). Overall, this study presents a comparative analysis of commercially available kits for the serodiagnosis of human VL and provides additional information on the prevalence of asymptomatic carriers in the Mediterranean basin. [ABSTRACT FROM AUTHOR]

Background: Leishmaniases are parasitic diseases caused by Leishmania species and transmitted by the bite of sand flies. The genus Lutzomyia and Phlebotomus of sand flies are known to be the responsible vector for transmitting almost all Leishmania species to humans. The detection of Leishmania DNA in species of the genus Sergentomyia, in different regions, suggests their likely role in Leishmania transmission. Methods: Our objective was to determine the potential geographical distribution of Sergentomyia minuta, the most dominant Sergentomyia species in Morocco, using ecological niche modeling. Results: The results showed the widespread geographical distribution of S. minuta in Morocco, specifically in northern and central Morocco where visceral and cutaneous leishmaniasis foci occur. There were six abiotic factors affecting the distribution of S. minuta whose annual precipitation, precipitation seasonality and precipitation of driest month as the most important ecological variables of the model. Conclusion: A positive statistical correlation between human leishmaniasis cases and S. minuta abundance was noted suggesting the potential involvement of S. minuta in local Leishmania transmission cycles. [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]

Background: Leishmaniasis is considered among the main endemic diseases in Morocco. However, further knowledge about epidemiological aspects of this disease is needed in several provinces to plan control and preventive strategies to tackle the disease. The present study aims to determine the epidemiological aspect of cutaneous and visceral leishmaniasis in Taza Province from 2007-2014 and to identify the circulating species in this province. Results: The temporal study from 2007 to 2014 showed that the number of cutaneous leishmaniasis cases increased since 2010. During the period of study, most leishmaniasis cases were detected in both urban and rural areas with 34% of cases detected in two urban localities, Bab Zitouna and Bab tété with 297 and 106 cases, respectively. The molecular study of cutaneous leishmaniasis showed the presence of non-sporadic Leishmania infantum and Leishmania tropica in this province. Regarding visceral leishmaniasis, Leishmania infantum is the species that has been identified. Conclusions: The epidemio-molecular study of leishmaniasis in Taza Province showed the coexistence of two species of Leishmania in the same foci. They also indicated that CL due to Leishmania infantum is more prevalent than reported in the literature. These results will be helpful for the implementation of control strategies by targeting dogs that constitute a reservoir of Leishmania infantum. [ABSTRACT FROM AUTHOR]

Background: Visceral leishmaniasis (VL) is a neglected parasitic disease that is fatal if left untreated. VL is endemic in Morocco and other countries in North Africa were it mainly affects children from rural areas. In Morocco, the direct observation of Leishmania parasites in bone marrow aspirates and serological tests are used to diagnose VL. Glucantime is the first line of treatment. The objective of this study was to report the costs associated to standard clinical management of pediatric VL from the provider perspective in Morocco. As a secondary objective we described the current clinical practices and the epidemiological characteristics of pediatric VL patients. Methods: From March to June 2014 we conducted a survey in eight hospitals treating pediatric VL patients in Morocco. A pro-forma was used to collect demographic, clinical and management data from medical records. We specifically collected data on VL diagnosis and treatment. We also estimated the days of hospitalization and the time to start VL treatment. Costs were estimated by multiplying the use of resources in terms of number of days in hospital, tests performed and drugs provided by the official prices. For patients receiving part of their treatment at Primary Health Centers (PHC) we estimated the cost of administering the Glucantime as outpatient. We calculated the median cost per VL patient. We also estimated the cost of managing a VL case when different treatment strategies were applied: inpatient and outpatient. Results: We obtained data from 127 VL patients. The median total cost per pediatric VL case in Morocco is 520 US$. The cost in hospitals applying an outpatient strategy is significantly lower (307 US$) than hospitals keeping the patients for the whole treatment (636 US$). However the outpatient strategy is not yet recommended as VL treatment for children in the Moroccan guidelines. VL diagnosis and treatment regimens should be standardized following the current guidelines in Morocco. [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: 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]