Geographical distribution and molecular survey of freshwater snail intermediate hosts of Schistosoma haematobium by DraI/Sh73 PCR in eliminated foci of Morocco.

Morocco had reached the level of schistosomiasis elimination 16 years ago. However the spread of freshwater snails in several breeding sites, and imported schistosome infection, still exist. Therefore, snail survey is a crucial component to sustain elimination progress. This study aimed to evaluate and to incorporate DraI/Sh73 PCR, for detecting early prepatent Schistosoma haematobium infection in snail host, into epidemiologic surveillance for schistosomiasis, particularly in reportedly eliminated foci where S.bovis overlaps with S. haematobium. The geographical distribution and the density of Bulinus truncatus and Planorbarius metidjensis were monitored for six years (2014-2019) and snail sampling were conducted in Fkih Ben Saleh province. All snails were analyzed in pools by DraI/Sh73 PCR. Results showed absence of Planorbarius metidjensi and none of the collected Bulinus truncatus snails were infected by S. haematobium. DraI/Sh73 PCR using pooled snail extracts is specific, feasible and suitable in routine malacological survey in the post elimination phase of schistosomiasis in Morocco.

Awareness of toxoplasmosis among postpartum women: a cross-sectional study in Morocco.

Introduction: awareness and knowledge of toxoplasmosis are particularly important, as an intervention point for the management of the disease. The aim of this study was to evaluate the awareness and knowledge regarding toxoplasmosis in a sample of postpartum Moroccan women. Methods: this was a cross-sectional descriptive survey carried out among 320 parturient at the National Reference Center for Neonatology and Nutrition at the Children's Hospital of Rabat. Results: of 320 parturient women responding to the survey, 227 (71%) had never heard about toxoplasmosis. While 18.1% of parturient stated knowing the transmission routes for toxoplasmosis. Regarding the transmission route, 53 (16.6%) women pointed at a domestic cat and 31 (9.7%) at eating raw or undercooked meat. Out of all participants, 60 (18.8%) women said they had received prevention advice during their pregnancy. The great majority (90%) of participants were unaware of the severity of the congenital infection. None of the participants showed a high level of knowledge about transmission routes, measures of prevention, and severity of CT. Conclusion: according to our survey, we observed that the majority of participants had never heard any information about toxoplasmosis. It is, therefore, necessary to educate women of childbearing age and pregnant women about the disease, especially concerning the transmission route and the prevention of infection and primary infection in non-immune women.

High performance of a novel point-of-care blood test for Toxoplasma infection in women from diverse regions of Morocco.

Point-of-care (POC) testing for Toxoplasma infection has the potential to revolutionize diagnosis and management of toxoplasmosis, especially in high-risk populations in areas with significant environmental contamination and poor health infrastructure precluding appropriate follow-up and preventing access to medical care. Toxoplasmosis is a significant public health challenge in Morocco, with a relatively heavy burden of infection and, to this point, minimal investment nationally to address this infection. Herein, we analyse the performance of a novel, low-cost rapid test using fingerstick-derived whole blood from 632 women (82 of whom were pregnant) from slums, educational centres, and from nomad groups across different geographical regions (i.e. oceanic, mountainous) of Morocco. The POC test was highly sensitive and specific from all settings. In the first group of 283 women, sera were tested by Platelia ELISA IgG and IgM along with fingerstick whole blood test. Then a matrix study with 349 women was performed in which fingerstick - POC test results and serum obtained by venipuncture contemporaneously were compared. These results show high POC test performance (Sensitivity: 96.4% [IC95 90.6-98.9%]; Specificity: 99.6% [IC95 97.3-99.9%]) and high prevalence of Toxoplasma infection among women living in rural and mountainous areas, and in urban areas with lower educational levels. The high performance of POC test confirms that it can reduce the need for venipuncture and clinical infrastructure in a low-resource setting. It can be used to efficiently perform seroprevalence determinations in large group settings across a range of demographics, and potentially expands healthcare access, thereby preventing human suffering.

Retrospective study on imported schistosomiasis in Morocco between 2005 and 2017.

Since 2004, no indigenous cases of schistosomiasis have been found in Morocco; only imported cases have been detected. The aim of the present study was to describe and analyse the epidemiological profile of imported schistosomiasis between 2005 and 2017, and, by this, attract attention to the probability of a reintroduction of this disease. During this period, 27 cases were recorded in Morocco, with a male predominance (13:1). All cases reported were found among African immigrants from Mauritania (37%), Mali (18%) and Senegal (15%). Schistosoma heamatobium was the most dominant specie. Most cases were reported in Rabat and Agadir, where there are many snail habitats. To prevent a re-emergence of the disease, the main challenge would be to consolidate and maintain a sustainable surveillance and control system of the importation of bilharzia. The frequency of asymptomatic schistosomiasis justifies a systematic health check-up for all travellers, migrants and immigrants.

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.

Global initiative for congenital toxoplasmosis: an observational and international comparative clinical analysis.

Globally, congenital toxoplasmosis remains a significant cause of morbidity and mortality, and outbreaks of infection with T. gondii represent a significant, emerging public health burden, especially in the developing world. This parasite is a threat to public health. Disease often is not recognized and is inadequately managed. Herein, we analyze the status of congenital toxoplasmosis in Morocco, Colombia, the United States, and France. We identify the unique challenges faced by each nation in the implementation of optimal approaches to congenital toxoplasmosis as a public health problem. We suggest that developed and developing countries use a multipronged approach, modeling their public health management protocols after those in France. We conclude that education, screening, appropriate treatment, and the development of novel modalities will be required to intervene successfully in caring for individuals with this infection. Gestational screening has been demonstrated to be cost-effective, morbidity-sparing, and life-saving. Recognition of the value and promise of public health interventions to prevent human suffering from this emerging infection will facilitate better patient and societal outcomes.

[Assessment of a supervision grid being used in the laboratories of cutaneous leishmaniasis in Morocco]. / 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.

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.

Selecting accurate post-elimination monitoring tools to prevent reemergence of urogenital schistosomiasis in Morocco: a pilot study.

BACKGROUND: After alleged stop of transmission of schistosomiasis and further down the line in post elimination settings, sensitive tools are required to monitor infection status to prevent potential re-emergence. In Rahala, where transmission cycle of Schistosoma haematobium is interrupted since 2004 but where 30% of snails are still infected by S. bovis, potential human S. bovis infection can't be excluded. As methods based on egg-counts do not provide the required sensitivity, antibody or antigen assays are envisaged as the most appropriate tools for this type of monitoring. METHODS: In this pilot study, the performances of three assays were compared: two commercially available antibody tests (ELISA and haemagglutination format) indicating exposure, and an antigen test (lateral flow strip format) demonstrating active infection. All 37 recruited study participants resided in Rahala (Akka, province Tata, Morocco). Participants had been diagnosed and cured from schistosomiasis in the period between 1983 and 2003. In 2015 these asymptomatic participants provided fresh clinical samples (blood and urine) for analysis with the aforementioned diagnostics tests. RESULTS: No eggs were identified in the urine of the 37 participants. The haemagglutination test indicated 6 antibody positives whereas the ELISA indicated 28 antibody positives, one indecisive and one false positive. ELISA and haemagglutination results matched for 18 individuals, amongst which 5 out of 6 haemagglutination positives. With the antigen test (performed on paired serum and urine samples), serum from two participants (cured 21 and 32 years ago) indicated the presence of low levels of the highly specific Schistosoma circulating anodic antigen (CAA), demonstrating low worm level infections (less than 5 pg/ml corresponding to probably single worm pair). One tested also CAA positive with urine. ELISA indicated the presence of human anti-Schistosoma antibodies in these two CAA positive cases, haemagglutination results were negative. CONCLUSIONS: To prevent reemergence of schistosomiasis in Morocco current monitoring programs require specific protocols that include testing of antibody positives for active infection by the UCP-LF CAA test, the appropriate diagnostic tool to identify Schistosoma low grade infections in travelers, immigrants and assumed cured cases. The test is genus specific will also identify infections related to S. bovis.

Geographical Distribution and New Situation of Leishmania Species after the Control of Cutaneous Leishmaniasis Foci in Errachidia Province, Morocco, in 2014.

In Errachidia province, the incidence of cutaneous leishmaniasis (CL) has increased over the past decade and it was higher in 2010 (860.34 per 100,000 inhabitants), with 3445 cases. The number of cases declined sharply and decreased from 3445 cases in 2010 to 8 cases in 2014 following the control action plan interventions. The total of patients was diagnosed only on clinical basis and the lesions were considered caused by L. major. The epidemiological study was conducted between 2001 and 2014 and the molecular detection of CL was studied to identify the circulating parasite species in this province by using the ITS1-PCR-RFLP methods. In 2010, the molecular identification of 11 samples revealed the presence of L. major in the most affected circles: Goulmima, Er-Rissani, and Errachidia. In 2014 the molecular characterization of 7 among 8 cases reported in this year showed the presence of L. tropica in Errachidia circle. This is the first molecular identification of L. tropica in Errachidia province. The detection of this species after the intensified control measures strategies suggests that it was probably dissipated through the dominance of L. major.

Effectiveness and Cost of Insecticide-Treated Bed Nets and Indoor Residual Spraying for the Control of Cutaneous Leishmaniasis: A Cluster-Randomized Control Trial in Morocco.

Cutaneous leishmaniasis (CL) remains an important public health problem in Morocco. A cluster-randomized trial was conducted with the following three study arms: 1) long-lasting insecticide-treated nets (LLINs) plus standard of care environmental management (SoC-EM), 2) indoor residual spraying (IRS) with α-cypermethrin plus SoC-EM, and 3) SoC-EM alone. Incidence of new CL cases by passive and active case detection, sandfly abundance, and cost and cost-effectiveness was compared between study arms over 5 years. Incidence of CL and sandfly abundance were significantly lower in the IRS arm compared with SoC-EM (CL incidence rate ratio = 0.32, 95% confidence interval [CI] = 0.15-0.69, P = 0.005 and sandfly abundance ratio = 0.39, 95% CI = 0.18-0.85, P = 0.022). Reductions in the LLIN arm of the study were not significant, possibly due to poor compliance. IRS was effective and more cost-effective for the prevention of CL in Morocco.

Field evaluation of alphacypermethrin in indoor residual spraying for leishmaniasis control in an endemic area, northern Morocco.

BACKGROUND: In Morocco, the main strategies of leishmaniasis vector control are based on environmental modifications. Use of local residual indoor spraying with synthetic pyrethroids is often envisaged. The need to evaluate this control method is essential. The current study was conducted to determine the efficacy of an alphacypermethrin IRS program against leishmaniasis vectors in an endemic area in the north of Morocco. METHODS: The survey was conducted in four neighbouring localities in three different districts in northern Morocco: Ait Chaib and Aichoun in Sefrou district, Bouassem (Boulmane) and Lmrouj (Taounate). Indoor residual spraying with alphacypermethrin at a dose of 30 mg/m2 was used in Ait Chaib and Lmrouj localities during 2010, 2011 and 2012, while localities of Aichoun and Bouassem were taken as control. In the four studied areas, sand flies were collected bimonthly from April to November in 2011 and 2012, using sticky traps, to determine their abundance and feeding pattern. Alphacypermethrin IRS were evaluated for their residual effect using the WHO cone bioassay test. Leishmaniasis incidence was estimated by passive and active case detection in each study area. RESULTS: Significant reductions in leishmaniasis incidence and in gravidity rate were observed when comparing sprayed and unsprayed localities. The residual activity of alphacypermethrin at the concentration used lasted 10 weeks after spraying. However, the abundance of sand flies was not significantly affected by alphacypermethrin IRS. CONCLUSION: This study indicated that IRS has a significant impact on leishmaniasis transmission; therefore it could be recommended as an effective tool for leishmaniasis control in areas with high leishmaniasis transmission.

Insecticide susceptibility status of Phlebotomus (Paraphlebotomus) sergenti and Phlebotomus (Phlebotomus) papatasi in endemic foci of cutaneous leishmaniasis in Morocco.

BACKGROUND: In Morocco, cutaneous leishmaniasis is transmitted by Phlebotomus sergenti and Ph. papatasi. Vector control is mainly based on environmental management but indoor residual spraying with synthetic pyrethroids is applied in many foci of Leishmania tropica. However, the levels and distribution of sandfly susceptibility to insecticides currently used has not been studied yet. Hence, this study was undertaken to establish the susceptibility status of Ph. sergenti and Ph. papatasi to lambdacyhalothrin, DDT and malathion. METHODS: The insecticide susceptibility status of Ph. sergenti and Ph. papatasi was assessed during 2011, following the standard WHO technique based on discriminating dosage. A series of twenty-five susceptibility tests were carried out on wild populations of Ph. sergenti and Ph. papatasi collected by CDC light traps from seven villages in six different provinces. Knockdown rates (KDT) were noted at 5 min intervals during the exposure to DDT and to lambdacyhalothrin. After one hour of exposure, sandflies were transferred to the observation tubes for 24 hours. After this period, mortality rate was calculated. Data were analyzed by Probit analysis program to determine the knockdown time 50% and 90% (KDT50 and KDT90) values. RESULTS: Study results showed that Ph.sergenti and Ph. papatasi were susceptible to all insecticides tested. Comparison of KDT values showed a clear difference between the insecticide knockdown effect in studied villages. This effect was lower in areas subject to high selective public health insecticide pressure in the framework of malaria or leishmaniasis control. CONCLUSION: Phlebotomus sergenti and Ph. papatasi are susceptible to the insecticides tested in the seven studied villages but they showed a low knockdown effect in Azilal, Chichaoua and Settat. Therefore, a study of insecticide susceptibility of these vectors in other foci of leishmaniasis is recommended and the level of their susceptibility should be regularly monitored.