Genetic Diversity and Population Structure of Leishmania infantum in Morocco as Revealed by Multilocus Sequence Typing (MLST) Approach.

Leishmania infantum is endemic in Morocco, and it causes both visceral (VL) and cutaneous leishmaniasis (CL). In this study, the multilocus sequence typing (MLST) approach was used to investigate the phylogeny and population structure of Leishmania infantum strains isolated from CL and VL patients and the canine reservoir in different leishmaniasis endemic foci in Morocco. For this purpose, eight loci (pgm, alat, me, fh, g6pd, pgd, gpi and cytb) were amplified in 40 samples, out of which 31 were successfully sequenced. The genetic diversity analysis detected a high degree of intraspecific genetic variability among the studied strains. The phylogenetic and the haplotype analyses showed that most of the strains from the same geographical areas clustered together. The recombination among Leishmania infantum strains was revealed through a splits tree analysis and the number of recombination events. Moreover, the assessment of the gene flow between Leishmania infantum and Leishmania tropica through phylogenetic analysis and haplotype diversity in two endemic foci where the two species were sympatric showed no genetic exchange between the two species.

Isolation, identification, and activity evaluation of antioxidant components from Inula viscosa: A bioguided approach.

Oxidative stress is linked to several invasive diseases which causes significant clinical and economic impact, therefore, there is a need to develop new antioxidants. The natural products could play an important role in overcoming the current need. In the present work, the antioxidant bioassay-guided fractionation of the ethanolic extract of Inula viscosa leaves (Asteraceae) was performed using DPPH and ABTS assays affording three known compounds, which were successfully characterized as ilicic acid (1), taxifolin (2) and quercetin (3) based on 1D, 2D NMR. Compounds 2 and 3 were identified as the most active, displaying similar or higher potency against ABTS (value 41.27 for quercetin and 142.58 for taxifolin) and similar activity against DPPH (value 41.27 for quercetin and 142.58 for taxifolin) than the well-known reference, ascorbic acid (value 65.36 for quercetin and 58.43 for taxifolin) but less potency than the standard gallic acid. The discussion of SAR of the antioxidant potential revealed that the type of natural product is crucial for the activity and the substitution pattern on the flavonoid skeleton modulate the antioxidant profile. Our findings show that I. viscosa leaves may be a natural source of antioxidants and once again the role of flavonoids health benefits is more strongly endorsed.

The Sporadic cutaneous leishmaniasis due to Leishmania infantum in Morocco: A presumably trend towards endemicity.

In Morocco, cutaneous leishmaniasis (CL) is an endemic disease; it is considered a major public health problem caused by three species Leishmaniamajor, Leishmaniatropica, and the dermotropic variant MON-24 of Leishmaniainfantum. This species has three incriminated vectors named; Phlebotomus ariasi, Phlebotomus longicuspis and Phlebotomus perniciosus, with the dog as reservoir.The main aim of this review is to elucidate the current epidemiological pattern of CL due to L.infantum and to investigate the factors facilitating its propagation throughout the country. Therefore, the number of CL cases due to L.infantum, their repartition; the distribution of L.infantum vectors, as well as the factors affecting their abundance and spread were investigated. We showed a wide extension of this form of CL, from the north of Morocco to the Saharan areas, as well as an increase of reported cases. This extension of the disease has been accompanied by a juxtaposed spread and a high abundance of confirmed vectors of L. infantum, which are present in almost all bioclimatic zones. In this review, we have highlighted the impact of climate: temperature, humidity, precipitation; vegetation and human activities on the geographical expansion of L. infantum vectors. These abiotic and biotic factors constitute favorable conditions for the increase of vector populations, and their introduction into areas where they did not exist before, and subsequently raise the risk of introduction of this form of cutaneous leishmaniasis into previously free areas. To conclude, CL by L.infantum, traditionally evolving as a sporadic form, is changing to an endemic mode, which seeks more epidemiological studies, and more attention from the health authorities when implementing control programs.

Bioguided Isolation of Active Compounds from Rhamnus alaternus against Methicillin-Resistant Staphylococcus aureus (MRSA) and Panton-Valentine Leucocidin Positive Strains (MSSA-PVL).

Despite intensified efforts to develop an effective antibiotic, S. aureus is still a major cause of mortality and morbidity worldwide. The multidrug resistance of bacteria has considerably increased the difficulties of scientific research and the concomitant emergence of resistance is to be expected. In this study we have investigated the in vitro activity of 15 ethanol extracts prepared from Moroccan medicinal plants traditionally used for treatment of skin infections. Among the tested species I. viscosa, C. oxyacantha, R. tinctorum, A. herba alba, and B. hispanica showed moderate anti-staphylococcal activity. However, R. alaternus showed promising growth-inhibitory effects against specific pathogenic bacteria especially methicillin-susceptible Staphylococcus aureus Panton-Valentine leucocidin positive (MSSA-PVL) and methicillin-resistant S. aureus (MRSA). The bioguided fractionation of this plant using successive chromatographic separations followed by nuclear magnetic resonance (NMR) and mass spectrometry (MS) including EIMS and HREIMS analysis yielded the emodin (1) and kaempferol (2). Emodin being the most active with MICs ranging between 15.62 and 1.95 µg/mL and showing higher activity against the tested strains in comparison with the crude extract, its mechanism of action and the structure-activity relationship were interestingly discussed. The active compound has not displayed toxicity toward murine macrophage cells. The results obtained in the current study support the traditional uses of R. alaternus and suggest that this species could be a good source for the development of new anti-staphylococcal agents.

Exploring the Anti-Infective Value of Inuloxin A Isolated from Inula viscosa against the Brain-Eating Amoeba (Naegleria fowleri) by Activation of Programmed Cell Death.

Primary amoebic meningoencephalitis (PAM), caused by the pathogenic free-living amoeba Naegleria fowleri, is a rare but fatal disease. Nowadays, no fully effective therapy is available to erradicate or prevent this disease. Natural products could constitute a promising source of useful bioactive compounds in drug discovery. The present study is a characterization of main active compounds from the ethanolic extract of Inula viscosa (Asteraceae) leaves against N. fowleri trophozoites. Four compounds (1-4) were successfully identified by spectroscopic techniques, but only inuloxin A displayed a potential antiamoebic activity with an IC50 of 21.27 µM. The specificity of this compound toward the studied strain leads us to analyze the insight into its mechanism of action by performing in vitro assays of programmed cell death markers and to discuss the structure-activity relationship (SAR). The obtained results demonstrated that inuloxin A interferes with various processes leading to membrane damage, mitochondria alteration, chromatin condensation, and ROS accumulation, which highlight features specific to apoptosis. The current findings could be a promising step for developing new effective drugs against PAM.

Sesquiterpenoids and flavonoids from Inula viscosa induce programmed cell death in kinetoplastids.

Neglected tropical diseases such as leishmaniasis and American trypanosomiasis represent an increasing health problem. Current treatments are not satisfactory which remains an urgent need for novel, cheap and safe chemotherapies. In the course of our ongoing search for new potential anti-protozoal agents, this study aimed to perform a bio-guided fractionation of Inula viscosa (Asteraceae) using in vitro assays against three strains of Leishmania and Trypanosma genus. Eight known compounds were identified from the ethanolic extract of leaves, sesquiterpenoids (3 and 4) and flavonoids (5 and 6) were characterized as the main bioactive constituents. Sesquiterpene lactones 3 and 4 (IC50 values between 4.99 and 14.26 µM) showed promising antiparasitic activity against promastigotes of L. donovani, L. amazonensis and epimastigotes of T. cruzi. Their structures were successfully characterized by spectroscopic techniques including 1D and 2D NMR experiments. Furthermore, the main bioactive compounds 4, 5 and 6 displayed higher potency (IC50 values between 0.64 and 2.13 µM) against amastigotes of L. amazonensis than miltefosine (IC50 3.11 µM), and a low toxicity on macrophages cell line (SI > 45). The analysis of structure-activity relationship (SAR) of the anti-protozoal activity revealed that lactonization or oxidation enhanced the biological profile, suggesting that the hydrophobic moiety was presumably involved in the activity by increasing the affinity and/or cell membrane permeability. In order to get an insight into the mechanism of action of these compounds, programmed cell death (PCD) experiments were performed, and the obtained results suggest that the reported compounds induced PCD in the treated parasites. These results highlight that sesquiterpenoids and flavonoids from I. viscosa could constitute an interesting scaffold for the development of novel antikinetoplastid agents.

From Wuhan to COVID-19 Pandemic: An Up-to-Date Review of Its Pathogenesis, Potential Therapeutics, and Recent Advances.

The emergence of a novel human coronavirus (SARS-CoV-2) causing severe contagious respiratory tract infections presents a serious threat to public health worldwide. To date, there are no specific antiviral agents available for this disease, currently known as COVID-19. Therefore, genomic sequencing and therapeutic clinical trials are being conducted to develop effective antiviral agents. Several reports have investigated FDA-approved drugs as well as in silico virtual screening approaches such as molecular docking and modeling to find novel antiviral agents. Until now, antiparasitic drugs such as chloroquine have shown the most relevant results. Furthermore, there is an urgent need to understand the pathogenesis of this novel coronavirus, its transmission routes, surface survival and evolution in the environment. So far, the scientific community has indicated a possible transmission of COVID-19 via blood transfusion which is challenging in the case of asymptomatic individuals. Protocols for pathogen inactivation are also needed. In this paper, we reviewed recent findings about this life-threatening pandemic.

Infectious Risk of the Hospital Environment in the Center of Morocco: A Case of Care Unit Surfaces.

BACKGROUND: Equipment and hospital surfaces constitute a microbial reservoir that can contaminate hospital users and thus create an infectious risk. The aim of this work, which was carried out for the first time at a hospital in Meknes (regional hospital in the center of Morocco), is to evaluate the microbiological quality of surfaces and equipment in three potential risk areas (burn unit, operating room, and sterilization service). METHODS: This study was carried out over a period of 4 months (February-May 2017). A total of 60 samples were taken by swabbing according to the standard (ISO/DIS 14698-1 (2004)) in an environment of dry area and equipment after biocleaning. Isolation and identification were performed according to conventional bacteriological methods and by microscopic observation for fungi. RESULTS: The study showed that 40% of surface samples were contaminated after biocleaning. The burn unit recorded a percentage of 70% contamination (p value <0.001), 13% for the sterilization service, and 7% for the operating room. 89% of the isolates were identified as Gram-positive bacteria against 11% for fungi (p value <0.001). Bacterial identification showed coagulase-negative staphylococci (32%), Bacillus spp. (16%), Corynebacterium (8%), and oxidase-negative Gram-positive bacillus (40%) while fungal identification showed Aspergillus niger (n = 2) and Aspergillus nidulans (n = 1). CONCLUSION: To control the infectious risk related to equipment and hospital surfaces, it would be necessary to evaluate the disinfection protocol applied in these units.

Antistaphylococcal Activity and Phytochemical Analysis of Crude Extracts of Five Medicinal Plants Used in the Center of Morocco against Dermatitis.

Novel drugs for methicillin-resistant Staphylococcus aureus (MRSA) hospital- and community-acquired infections are needed because of the emergence of resistance against antibiotics. In this study, methanolic and aqueous extracts of Berberis hispanica, Crataegus oxyacantha, Cistus salviifolius, Ephedra altissima, and Lavandula dentata selected from an ethnopharmacological study to treat skin infections in Sefrou city (Center of Morocco) were tested for their antistaphylococcal activity against strains often involved in cutaneous disorders: two methicillin-resistant Staphylococcus aureus strains and one strain of Staphylococcus epidermidis using the well-diffusion assay, while the agar macrodilution method was used to determine the minimal inhibitory concentrations. The total phenolic compounds and flavonoid contents of all tested extracts were also evaluated. Three of the five methanolic extracts showed an important antibacterial activity. Berberis hispanica extract was the most active with a minimal inhibitory concentration of 04.00 mg/ml against all tested strains, followed by Cistus salviifolius and Crataegus oxyacantha extracts containing the highest amounts of total phenols (133.83 ± 9.03 and 140.67 ± 3.17 µg equivalent of gallic acid/mg of extract). However, the aqueous extracts have not shown any activity against the tested strains. The current data suggested that the most active extracts can be a good source of natural antistaphylococcal compounds and warrants further investigations to isolate bioactive molecules.

Circulating species of Leishmania at microclimate area of Boulemane Province, Morocco: impact of environmental and human factors.

BACKGROUND: Cutaneous leishmaniasis (CL) is widely distributed in Morocco where its geographical range and incidence are related to environmental factors. This study aimed to examine the impact of several factors on the distribution of CL in Boulemane Province, which is characterized by several microclimates, and to identify the Leishmania species circulating in these areas. METHODS: Ordinary least squares regression (OLSR) analysis was performed to study the impact of poverty, vulnerability, population density, urbanization and bioclimatic factors on the distribution of CL in this province. Molecular characterization of parasites was performed using a previously described PCR-RFLP method targeting the ITS1 of ribosomal DNA of Leishmania. RESULTS: A total of 1009 cases were declared in Boulemane Province between the years 2000 and 2015 with incidences fluctuating over the years (P = 0.007). Analyzing geographical maps of the study region identified four unique microclimate areas; sub-humid, semi-arid, arid and Saharan. The geographical distribution and molecular identification of species shows that the Saharan microclimate, characterized by the presence of Leishmania major was the most affected (47.78%) followed by semi-arid area where Leishmania tropica was identified in three districts. Among several environmental factors included in the study, poverty had the greatest influence on the spatial extension of the disease in this province. CONCLUSIONS: The incidence of CL in Boulemane Province varies between microclimate areas, and environmental factors partly explain this variation. However, the existence of CL in the most affected districts is mainly related to poverty, population movement and human behavior. To our knowledge, this the first study utilizing molecular techniques to confirm L. tropica and L. major as the causative agents of CL in Boulemane Province. Our findings indicate that the spatial and temporal distribution of CL in Boulemane Province is strongly related to poverty and population movement.

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.

Staphylococcus aureus nasal carriage in hemodialysis centers of Fez, Morocco.

BACKGROUND AND OBJECTIVES: Staphylococcus aureus (S. aureus) nasal carriage may be responsible for some serious infections in hemodialyzed patients. The main target of this study was to estimate the prevalence of S. aureus nasal carriage in hemodialysis outpatients and medical staff in hemodialysis centers specifically in Fez region. The second target is to identify the risks of colonization, resistance pattern of isolates and their virulence toxin genes. PATIENTS AND METHODS: Nasal swab specimens were obtained from 143 hemodialyzed outpatients and 32 medical staff from January to June 2012. Each participant completed a short questionnaire. Nasal carriage of S. aureus was demographically related (age, gender, hemodialysis duration), comorbidity (diabetes, malignancy) and exposure to health care (dialysis staff, hospitalization). PCR (Polymerase Chain Reaction) were used on all the isolates in the research of twelve staphylococcal enterotoxins genes. Also the PCR was used to investigate on the three factors epidermal cell differentiation inhibitors; three exfoliatin toxins; two leukotoxins; the toxic shock syndrome toxin-1 and the hemolysin beta genes. RESULTS: Nasal screening revealed 38.16%, 50% and 18.75% S. aureus carries in chronic, acute hemodialysis patients and medical staff, respectively. Only young participants were likely to be S. aureus carries (p = 0.002). But there were no gender differences between the isolate carriers and non-carriers or some comorbidity factors such as viral hepatitis B and C, HIV (Human Immunodeficiency Virus) infections, diabetes, chronic smoking, recent hospitalization or antibiotic therapy. Out of all isolates, only one (1.61%) was methicillin-resistant and Twenty-one (33.87%) had at least two virulence toxin genes. CONCLUSIONS: Knowledge and monitoring of antibiotic resistance profile and virulence of S. aureus carriage are essential in the treatment of infections generated by this pathogen, as well as in the control of clonal dissemination and prevent the spread of S. aureus resistance.