Moroccan lymphocyte subsets reference ranges: age, gender, ethnicity, and socio-economic factors dependent differences.

Lymphocyte subsets reference ranges are helpful for a precise diagnosis and therapy of various diseases. We attempted in the current study to establish Moroccan lymphocyte reference range and reveal age, gender, ethnicity, income, and instructional levels dependent differences. Lymphocyte subsets percentage and absolute count were determined by 4-color flow cytometry in a population study of 145 adults Moroccan healthy volunteers. Analysis showed significant age-dependent changes. Age was associated with a decrease of naïve CD4+ and CD8+ T cells and an increase of memory CD4+ or CD8+ T cells. Activated CD4+ CD38+ and CD8+ CD38+ T cells, Treg as well as NK cell showed age-dependent alterations. In contrast, B cells remained unchanged. A higher percentage of CD3+ and CD4+ T cells was observed in females while CD8+, B and NK cells count were higher in men. Ethnicity, instructional levels, and personal income seem to not influence lymphocyte subsets reference values. This study provides reference ranges for lymphocyte subsets of healthy Moroccan adults. These results can be used for other North African (Maghrebian) countries considering their geographic, ethnic, economic, and cultural similarities.

Genetic diversity in Leishmania infantum and Leishmania tropica isolates from human and canine hosts in northern Morocco.

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.

Toxoplasma gondii seroprevalence among pregnant women in Rabat, Morocco.

BACKGROUND: Toxoplasmosis is an infectious disease caused by a protozoan parasite named Toxoplasma gondii (T.gondii). Pregnant women are considered one of the risk groups. The objective of this retrospective study is to provide an updated estimate of the seroprevalence of anti-T. gondii antibodies among a group of Moroccan pregnant women monitored at the Parasitology Laboratory of the National Institute of Hygiene in Rabat in Morocco. METHODS: Serum samples were tested for the presence of specific anti-T. gondii immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies using indirect enzyme-linked immunosorbent assay (ELISA). Anti-Toxoplasma IgM- and IgG-positive cases were also evaluated with the anti-Toxoplasma IgG avidity test. All cases were evaluated according to the age, parity, and historical of abortion. RESULTS: Among 677 pregnant women, 94.1% (637/677) were serologically screened for the first time and therefore had no knowledge of their serological status, and only 5.9% (40/677) were screened for the second or third time. The overall anti-T. gondii IgG and IgM seropositivity among the 637 pregnant women included in the study analysis was 43% (274/637) and 3.9% (25/637), respectively. The use of the IgG avidity test allowed excluding recent infection among 83% of cases with IgG and IgM positive sera. The mean age was 29.4 ± 6.3 years. The result of the bivariate analysis revealed that the age influenced significantly the seroprevalence rate, while the parity and the existence of previous spontaneous abortion did not have any significant statistical correlation with seropositivity to T. gondii. CONCLUSION: This study shows that 43% of pregnant women were positive and 57% of them had no antibody against the T. gondii infection. However, the pregnancy follow-up and the counseling of pregnant women remain essential for the prevention of congenital toxoplasmosis.

The Level of Knowledge about Toxoplasmosis among University Students in Rabat in Morocco.

The aim of this descriptive cross-sectional study is to evaluate the knowledge of toxoplasmosis among medical, biology, and veterinary students in Rabat in Morocco. The data was collected by using a questionnaire which includes demographic characteristics, epidemiology, diagnosis, and clinical issues related to knowledge of toxoplasmosis. During analysis, the study groups were divided based upon their specialty of students who were medical, biology, and veterinary students. Out of 230 students, 55.2% were female and 44.8% were male. The average age of the study population is 21.7 ± 02 years. Less than half (42.6%) have heard of the disease; most of them have heard from faculty during studies in classrooms with 75.8%, and 3.2% were from the internet. Only 36.5% knew the correct causative agent of toxoplasmosis, and 32.1% were aware of the definitive host. The current study documented that there are gaps in the knowledge of the students regarding toxoplasmosis. Therefore, the present study puts the basis for future studies highlighting the importance of educating students to improve knowledge and attitudes towards toxoplasmosis.

Inflammatory Nasal Swelling due to Leishmania tropica.

Since its discovery in the 19th century, cutaneous leishmaniasis has been a major public health problem, especially with the appearance of more and more unusual cases of cutaneous lesions due to this parasite. Indeed, the present study joins the previous studies and describes a typical case of a nasal lesion due to Leishmania infection. This is a 20-year-old young man, with no particular pathological history, from an epidemic focus who presented with inflammatory nasal swelling similar to a mucocutaneous form. However, the X-ray data showed that no lysis of the bones proper to the nose was detected and no damage to the underlying mucosa was observed. Nevertheless, the parasitological diagnosis confirmed the presence of amastigotes, and the results of the molecular study showed that the electrophoretic profile was comparable to that of L. tropica. After diagnosis and confirmation, treatment with meglumine antimonate at the rate of two ampoules/injection (one ampoule = 5 ml) of antimony salt for one month was administered intramuscularly with favorable outcome. Atypical forms of cutaneous leishmaniasis constitute a rare and unusual entity often leading to diagnostic delay. For this, the clinical examination must take into account both exceptional presentations of Leishmania infection, in particular in subjects living or having stayed in an endemic area, in order to ensure appropriate and early treatment.

Control of cystic echinococcosis in the Middle Atlas, Morocco: Field evaluation of the EG95 vaccine in sheep and cesticide treatment in dogs.

BACKGROUND: Cystic echinococcosis (CE) is an important cause of human morbidity and mortality worldwide, particularly in Morocco and other North African countries. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the potential of three strategies to reduce Echinococcus granulosus transmission: (1) 4-monthly treatment of dogs with praziquantel, (2) vaccination of sheep with the EG95 vaccine and (3) a combination of both measures. These measures were implemented during four consecutive years in different areas of the Middle Atlas Mountains in Morocco. The outcome of the interventions was assessed through hydatid cyst (viable and non-viable) counts in liver and lungs using necropsy or in vivo ultrasound examination of the liver. A total of 402 lambs were recruited for annual vaccination with the EG95 anti-E. granulosus vaccine and 395 similar lambs were selected as non-vaccinated controls. At approximately four years of age the relative risk (estimated as odds ratio) for vaccinated sheep to have viable hydatid cysts compared with non-vaccinated controls was 3% (9.37% of the vaccinated sheep were found infected while 72.82% of the controls were infected; p = 0.002). The number of viable cysts in vaccinated animals was reduced by approximately 97% (mean counts were 0.28 and 9.18 respectively; p<0.001). An average of 595 owned dogs received 4-monthly treatment during the 44 months trial, corresponding to 91% of the owned dog population. Approximately, 5% of them were examined for E. granulosus adult worms by arecoline purge or eggs in feces (confirmed by PCR). The proportion of infected dogs significantly decreased after treatment (12% versus 35%; p<0.001). Post-treatment incidence of re-infestation corresponded to a monthly risk of 4% (95% CI: 3-6%). Treatment of owned dogs on a 4-monthly basis did not reduce the level of transmission of E. granulosus to sheep, nor did it enhance the level of control generated by vaccination of sheep with EG95, possibly because of unowned dogs and wild canids were not treated. CONCLUSIONS/SIGNIFICANCE: These data suggest that vaccination of sheep with EG95 has the potential to reduce the level of CE in Morocco and in other parts of the world with similar transmission dynamics. Under the epidemiological circumstances existing in the trial area, 4-monthly treatment of owned dogs with praziquantel was insufficient to have a major impact of E. granulosus transmission to sheep.

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.

The first report of the evaluation of the knowledge regarding toxoplasmosis among health professionals in public health centers in Rabat, Morocco.

BACKGROUND: The assessment of the knowledge of Toxoplasma gondii infection among health professionals is essential to design an effective management strategy. The current study was conducted to assess the knowledge and perception of health professionals working in urban public health centers of different parts of Rabat in Morocco. METHODS: A cross-sectional study was conducted from March 15 to June 15, 2017, in urban public health centers selected in the prefecture of Rabat in Morocco. A structured questionnaire was completed by participants and included questions on the epidemiology and diagnosis of toxoplasmosis and clinical issues related to the infection. RESULTS: Ninety-six health professionals participated, including medical doctors, nurses, midwives, and laboratory technicians. Most of them were female (86, 89.58%). The mean age was 40.51 ± 10.26 years, and the mean length of time working in the field of healthcare was 15.92 ± 8.55 years. Eighty one percent (86, 81.25%) of health professionals knew the agent of toxoplasmosis, and 62 (64.5%) knew the definitive host of the parasite. Regarding clinical symptoms, 55 (57.29%) of the respondents knew that toxoplasmosis is an asymptomatic disease in immunocompetent persons. More than half of the respondents correctly identified the main routes of transmission: eating raw or undercooked meats, unwashed fruits and vegetables, and having direct contact with cats. However, only 29 (30.21%) of them believed that water can be a risk factor for the transmission of toxoplasmosis. Regarding diagnosis, only 14 (14.58%) health professionals knew about the avidity test. CONCLUSIONS: The implementation of educational interventions is recommended to raise awareness of toxoplasmosis among health professionals who provide prenatal care in public health centers.

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.

Risk Factors Associated with Leishmaniasis in the Most Affected Provinces by Leishmania infantum in Morocco.

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.

Cystic Echinococcosis in Three Locations in the Middle Atlas, Morocco: Estimation of the Infection Rate in the Dog Reservoir.

A longitudinal study was carried out in Middle atlas, Morocco (locality of Had Oued Ifrane) in a population of 255 dogs from three localities, including two categories of dogs (owned and stray dogs). The dogs were investigated three times over a period ranging from 4 to 8 months between December and August. At each investigation, dogs were treated with arecoline, inducing defecation and allowing feces collection. Dogs were further treated with praziquantel to clear them from Echinococcus granulosus. Microscopic examination of feces was performed to assess the infection status of dogs at each investigation, and positive samples underwent copro-PCR to determine the circulating strain of E. granulosus. A high prevalence of infestation ranging from 23.5% to 38.8% and from 51.3% to 68.5% was, respectively, found in owned and in stray dogs. The PCR results revealed the presence of G1 strain in all positive samples. A logistic regression model was used to determine the incidence of infestation and showed that stray dogs underwent a significantly higher risk of infection (odds ratio = 14; 95% confidence interval: 6-30; p < 0.001) compared with owned dogs. Only anthelmintic treatment intervals of 2 months efficiently prevented egg shedding in owned and stray dogs. The seasonal effect was also significant, with the highest risk of reinfestation in winter and the lowest risk in summer. This study confirms that stray dogs undergo an increased risk of infestation by E. granulosus and indicate that infective pressure is influenced by season.

Leishmaniasis in Northern Morocco: Predominance of Leishmania infantum Compared to Leishmania tropica.

In Morocco, Leishmania infantum species is the main causative agents of visceral leishmaniasis (VL). However, cutaneous leishmaniasis (CL) due to L. infantum has been reported sporadically. Moreover, the recent geographical expansion of L. infantum in the Mediterranean subregion leads us to suggest whether the nonsporadic cases of CL due to this species are present. In this context, this review is written to establish a retrospective study of cutaneous and visceral leishmaniasis in northern Morocco between 1997 and 2018 and also to conduct a molecular study to identify the circulating species responsible for the recent cases of leishmaniases in this region. Data concerning leishmaniases cases were collected from the Epidemiology and Disease Control Directorate from 1997 to 2018. Human samples obtained from peripheral laboratories were examined using PCR-ITS1 method. The ITS1 products were subjected to digestion with the restriction endonuclease Mn1-I. Between 1997 and 2018, a total of 1,255 cases of cutaneous and visceral leishmaniasis were recorded in Tangier-Tetouan-Al Hoceima Region, i.e., 1.56% of the reported cases in Morocco (1,255/80,299). Concerning the geographical study covering the period 2007-2018, 79.5% (105/132) of the sectors were affected by leishmaniases. The molecular results showed that Humans were found to be infected with the L. infantum species with a high infection rate compared to L. tropica infection. Moreover, molecular characterization using ITS1 PCR-RFLP showed that the density of L. infantum was significantly higher (n = 68/81; 84%) than that of L. tropica (n = 13/81; 16%) (P-value 9.894e-10). While regarding visceral leishmaniasis, L. infantum was the only species responsible of this form. These findings of this study showed the emergence of L. infantum in Morocco and suggest that this species might be more prevalent than previously thought. Furthermore, the molecular determination of L. infantum will be helpful for control strategies by taking into consideration the reservoir of this species.

A review of cutaneous leishmaniasis in Morocco: A vertical analysisto determine appropriate interventions for control and prevention.

Leishmaniasis is considered one of the most neglected diseases worldwide. In Morocco, cutaneous leishmaniasis is an important public health problem. Leishmania major and Leishmania tropica are the two major species in this country. Despite all efforts, monitoring and control of the cutaneous leishmaniasis is still challenging. We used for the first time a vertical analysis of the control of cutaneous leishmaniasis in Morocco from the document review and publications. This analysis allowed us to develop an epidemiological model that emphasized key possible interventions. No evaluation studies of these interventions in Morocco were done. Global Evidence underline the effectiveness of preventive interventions produced in integrate inter-sectorial strategy framework (e.g use of insecticide-treated bednets, indoor residual spraying and rodents' control) rather than treatments such as based thermotherapy, cryotherapy, photodynamic therapy, CO2 laser and paromomycin. Therefore, integrated vector management control (IVMC) with communityc participation is recommended as effective strategy. Strengthening of the IVMC with community involvement are necessary conditions to improve the program of cutaneous leishmaniasis and prevent epidemic foci appearance.

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.

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.

Serodiagnosis of Toxoplasmosis: The effect of measurement of IgG avidity in pregnant women in Rabat in Morocco.

BACKGROUND: The diagnosis of Toxoplasmosis in pregnant women during the early first trimester of pregnancy is very important for preventing congenital infection of the fetus; it will not only prevent the risk of transmitting the infection to the fetus but it will also enable to give these women a preventive treatment. In this study, the avidity test was performed on pregnant women during their first prenatal visit at the National Institute of Hygiene in Rabat, Morocco. FINDINGS: One hundred and twenty-eight sera samples were collected from 128 pregnant women between August 2015 and June 2016; these women were chosen retrospectively and were in their first four months of pregnancy. The samples were screened using the specific anti-Toxoplasma IgG and IgM antibodies and were subjected to an IgG avidity test. After the serological screening, only 54 women (42.4%) were tested positive for IgG antibodies and five women (3.9%) were tested positive for both anti-Toxoplasma IgG and IgM antibodies. Four IgM-negative women had low-avidity antibodies. However, none of the IgG-avidity test had detected low-avidity antibodies in the five IgM-positive women; three women (60%) had high-avidity antibodies, indicating that the infection was acquired in the distant past. CONCLUSION: The avidity test is a helpful tool to exclude a recently acquired toxoplasmosis infection within IgM-positive serum samples in pregnant women during their first trimester of pregnancy. Thus, allowing to perform an appropriate therapeutic intervention.

A case control study of risk factors associated with pulmonary tuberculosis in romania: experience at a clinical hospital of pneumology.

BACKGROUND AND AIM: Tuberculosis (TB) remains a major public health issue in Romania. The aim of the present study was to evaluate the potential demographic, socioeconomic and behavioral risk factors for TB among hospitalized patients in Romania. METHODS: This is a case-control study conducted between March 1st 2014 and March 30th 2015 at Leon Daniello Clinical Hospital of Pneumology, Cluj Napoca. A total of 150 TB patients defined as "cases" were matched for age, sex and county of residence to 150 controls selected from patients attending the same hospital with respiratory diseases other than TB. Data collection was obtained through patient interviews using a structured questionnaire. Factors potentially associated with TB infection were analyzed using univariate and multivariate logistic regression. RESULTS: Factors independently associated with TB were illiteracy (OR=2.42, 95% CI 1.09-5.37), unemployment (OR=2.08, 95% CI 1.23-3.53), low household income (OR=4.12, 95% CI 2.53-6.71), smoking (more than 20 cigarettes per day) (OR=2.12, 95% CI 1.20-3.74), poor knowledge of TB (OR=3.46, 95% CI 1.97-6.07), presence of TB patient in household (OR=4.35, 95% CI 1.42-13.36), prior TB treatment (OR=2.2, 95% CI 1.93-2.5) and diabetes (OR=3.32, 95% CI 1.36-8.08). CONCLUSION: This study provided useful information that might help to develop and adapt effective policies for TB control in Romania.

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.

[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.

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.