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Introduction: Cutaneous leishmaniases (CL), a wide range of cutaneous diseases caused by diverse species of Leishmania genus parasites, are among the most neglected infectious diseases. While they are non-fatal, CL are highly morbid with disfiguring lesions, which could be chronic, leaving lifelong unsightly scars; they are combined with psychological distress and social stigma. The efficiency of treatment highly depends on the infecting Leishmania species. Diagnosis is mainly based on microscopic direct examination (DE) of Giemsa-stained smears needing experienced microscopists. It can be laborious and time-consuming when the parasite load is low. DE is poorly sensitive and does not identify Leishmania species. So far, only DNA assays accurately identify the species. Despite their wide use for generic detection, PCR methods also require equipment and additional steps to identify causal Leishmania species. L. major is hyperendemic in many countries in Africa, the Middle East, and Asia, where other species co-occur with different endemicity levels according to the situations. This complicates disease management and treatment, particularly as distribution and epidemiology of leishmaniases remain poorly understood. Here, we aimed for a simple and rapid molecular diagnostic test to detect and identify L. major , a predominant CL causal species, which could be prone to become a control tool at the point of care, in endemic areas, using isothermal recombinase DNA amplification (recombinase polymerase amplification, RPA, or recombinase aided amplification, RAA) coupled to detection by the lateral flow (LF) chromatography on a PCRD cassette. Methods: To develop an L. major species-specific RPA-LF assay, computational analysis of 70 Leishmania DNA targets, identified through bibliography and database searches, selected five targets. We designed and tested 7 primer pairs/probe sets to specifically amplify L. major DNAs. First, the primers were tested for species specificity and sensitivity using basic RPA chemistry. Then, to develop RPA-coupled LF detection, we shifted to the nfo chemistry. Results: This way, we retained one set for further investigation, which confirmed it is L. major species-specific. Tested on 86 human cutaneous samples, this selected set was able to detect 100% of L. major infections in confirmed CL patients. We did not observe any cross-reactivity with lesions due to L. infantum or L. tropica. [ABSTRACT FROM AUTHOR]

Dermatology is an important specialty in veterinary medicine, focusing on the skin and its appendages. Therefore, the present study highlighted the percentage of skin disorders, as well as their associated risk factors, from cases received at the Parasitology-Dermatology clinic of the Hassan II Institute of Agronomy and Veterinary Medicine (IAV Hassan II), Rabat, Morocco for two years. A total of 1561 dogs (1450 dogs were in 28 different pure breeds and 111 dogs were mongrels, 805 males, and 756 females, with an average age of 6.5 years old) were presented at the University Veterinary Teaching Hospital (UVTH) of the IAV Hassan II from the end of October 2020 to the end of May 2022 (including vaccinations) and 125 dogs were assessed and 161 skin diseases were found (a few dogs had more than one skin disease). Dermatological examinations represented an average of 8.00% (125/1561) of all canine cases received at the University Veterinary Teaching Hospital. The most common clinical signs were pruritus, alopecia, erythema, onychogryphosis, and visible ectoparasites. Parasitic dermatoses were the most frequent, representing 44.10% of all dermatological cases, followed by allergic dermatoses (25.47%) and fungal skin infections (19.25%). Bacterial skin infections and dermatological manifestations of endocrine disorders were infrequent, representing 8.70% and 2.48%, respectively of all observed cases. Risk factors contributing to the occurrence of canine skin disorders included age and lifestyle for sarcoptic mange. An apparent predilection for the living environment was observed in the case of canine leishmaniosis, and an apparent predilection for sex regarding otodectic mange was also demonstrated. Similar results were found for the living environment and lifestyle concerning canine atopic dermatitis. Data reported herein fill gaps in knowledge of skin disorders and their associated risk factors in dogs in Morocco, demonstrating the dominance of skin diseases of zoonotic interest, including flea bite allergy dermatitis (FBAD), dermatophytosis, and canine leishmaniosis. [ABSTRACT FROM AUTHOR]

Cutaneous Leishmaniasis (CL) is a major global health problem requiring appropriate diagnosis methods. Its diagnosis is challenging, particularly in resource-limited settings. The integration of Artificial Intelligence (AI) into medical diagnostics has shown promising results in various fields, including dermatology. In this systematic review, we aim to highlight the value of using AI for CL diagnosis and the AI-based algorithms that are employed in this process, and to identify gaps that need to be addressed. Our work highlights that only a limited number of studies are related to using AI algorithms for CL diagnosis. Among these studies, seven gaps were identified for future research. Addressing these considerations will pave the way for the development of robust AI systems and encourage more research in CL detection by AI. This could contribute to improving CL diagnosis and, ultimately, healthcare outcomes in CL-endemic regions. [ABSTRACT FROM AUTHOR]

Background and Aims: Leishmaniasis is a neglected tropical infection caused by Leishmania parasite that affect human and animal. In Morocco, the cutaneous leishmaniasis has spread substantially to the new areas. The surveillance limited to active foci may underestimate the occurrence of cutaneous leishmaniasis (CL). This study aims to investigate the local transmission of CL in rural districts of Youssoufia province, central Morocco, as a potential focus of CL. Methods: For this purpose, parasitological, molecular and entomological investigations were carried out in this area. Data collection concerns potential vectors and human cases. Thus, 402 patients were examined for suspected leishmaniasis lesions in three localities of the province of Youssoufia. In these same localities, 983 sand flies were collected by CDC light traps and sticky paper during one‐night per month during 6 months. These sand flies were all identified morphologically using the Moroccan identification key. Results: The results showed that among the 25 skin lesions detected in a population of 402 individuals, 18 were confirmed by kDNA nested PCR as CL positive patients, of which only 25% were positive by direct examination. Leishmania tropica and Leishmania major were identified as causative agents of CL in the study area. Direct parasitological examination showed a low sensitivity (27.78%), especially for L. major, although its specificity was evaluated at 100%. Regarding entomological results, both genera of the Moroccan sand fly were collected in the study area: Genus/Phlebotomus (75.28%) and Sergentomyia (24.72%). Phlebotomus (P) papatasi, the proven vector of L. major, was the most abundant species (33.98%), followed by Paralongicollum sergenti (22.58%), the confirmed vector of L. tropica; while Sergentomyia (S) minuta, P. longicuspis, S. fallax and P. kazeruni were collected with, respectively, 17.60%, 16.99%, 7.12% and 1.73%. Conclusion: This study constitutes the first report of CL in the study areas, as well as the coexistence of L. tropica and L. major in these rural localities. Local transmission of CL is highly probable, as indicated by the prevalence of the two proven vectors of L. major and L. tropica. To control the spread of this disease, our results suggest the use of highly sensitive molecular methods to detect CL cases in potential leishmaniasis foci, which will improve surveillance. [ABSTRACT FROM AUTHOR]

In Morocco, leishmaniases are a major public health problem due to their genetic diversity and geographical distribution. Cutaneous leishmaniasis (CL) is an infectious disease caused by various species of Leishmania and transmitted typically by bite of phlebotomine sand flies. This study identifies sand fly fauna in Ibaraghen village, province of Azilal, which is a focus of CL, by combination of morphological and molecular methods (sequencing of COI gene, MALDI-TOF MS protein profiling). Nested-kDNA PCR was used to detect and identify Leishmania species within potential vector species. 432 CDC light traps were placed at different heights above ground level at four capture sites during a whole year. Traps at 1.5 m above the ground yielded capture of sand flies almost double compared to above ground level (29.33%), while the collection reached 55.09% when the traps were placed 2.5 m above ground. A total of 2,830 sand flies were collected, 2,213 unfed specimens were morphologically identified, 990 males (44.73%) and 1,223 females (55.26%) of 13 species; ten Phlebotomus species and three Sergentomyia species. Six species were analysed by MALDI-TOF MS protein profiling (4 Phlebotomus and 2 Sergentomiya species), and their identification was confirmed by COI sequencing. 1,375 unfed females were screened for the presence of Leishmania by nested-kDNA PCR in pools, 11/30 pools of P. sergenti showing a single band of 750 bp corresponding to L. tropica. Our results confirm the role of P. sergenti as a proven vector in Azilal focus of cutaneous leishmaniasis; however, the relative abundance of other species known as vectors of Leishmania species emphasizes the risk of introduction of L. infantum and L. major in this province. For the first time in Morocco, a combined approach to identify sand flies by both morphology and molecular methods based on DNA barcoding and MALDI-TOF MS protein profiling was applied. [ABSTRACT FROM AUTHOR]

An appreciated sampling technique is essential for achieving optimum results from diagnostic tests of cutaneous leishmaniasis (CL); conventional sampling methods, such as skin biopsy and dermal scrapping, are painful for the patients and require qualified staff and hospital facilities, while swabbing is patient‐friendly more comfortable than invasive traditional techniques and can be carried out under field conditions. The aim of this study is to evaluate a non‐invasive sampling method (swab) in the cutaneous leishmaniasis diagnosis, compared to microscopic examination. The recruitment of 205 patients was done during 3 years in six regions known as endemic CL foci in the south‐east, the centre, the south‐west and the north of Morocco. The results showed that molecular detection of the nuclear marker ITS1 on swab materials is to be less sensitive than microscopic examination, and the difference was statistically significant (p‐value =.036). Thereafter, 55 patients were randomly selected to compare the results of two molecular techniques (ITS1‐PCR and nested KDNA‐PCR), performed both on swab and on stained smears used for microscopic examination; ITS1‐PCR results from stained smears reached 87% positivity against 65,2% for cotton swab; and it was statistically significant (p‐value =.019); on the other hand for the KDNA marker, results from cotton swab reached 93% of positivity against 91% for stained smears; and statistically the difference was not significant (p‐value =.5113). One can presume that rubbing over the lesion with cotton swab implies a low parasitic load collection, but the choice of the amplification target greatly influences the results obtained from cotton swab; on top of that swab remains useful in the cases of patients with multiples lesions or the latter are located in sensitive places difficult to reach with an invasive method such as the eyelids, the lips or other mucous areas. [ABSTRACT FROM AUTHOR]

Summary: In Morocco, cutaneous leishmaniasis (CL) caused by Leishmania tropica is endemic to locations where the predominantly anthropophilic blood‐feeding Phlebotomus sergenti and humans co‐perpetuate. The objective of this study was to explore whether the range of epidemiological features of CLcould be linked to the range of L. tropica genetic heterogeneity and to further explore the relationships between the genetic diversity of L. tropica in both P. sergenti and humans. L. tropica DNAwas extracted from dermal scarping smears of 125 CLpatients. Genetic polymorphisms were analyzed by sequencing the internal transcribed spacer (ITS) 1 and 5.8S rDNAgene. Nucleotide diversity (π), haplotype diversity (Hd) and Tajima's D test for neutrality, as well as overall and pairwise FSTvalues, were calculated using Arlequin ver 3.5 software. Out of the 125 amplified DNAsequences, 93 were completely sequenced and 13 L. tropica haplotypes were identified, which confirmed the significant genetic heterogeneity of L. tropica in Morocco. The most common haplotype included 74 out of 93 sequences; this haplotype is not only widely represented but was also detected in P. sergenti, which is known to be the most abundant species in the studied foci. Considering the negative value calculated using Tajima's D index, we briefly discussed the hypothesis that the L. tropica common haplotype propagation could be a sign of its fitness in P. sergenti and human hosts. Furthermore, analysis of molecular variance (AMOVA) shows significant correlations between intraspecific variants of L. tropica and patients' geographic origins. The long‐term goals of the present pilot study are to further explore the relationships between the genetic diversity of L. tropica in human and P. sergenti populations. [ABSTRACT FROM AUTHOR]

BACKGROUND: Arboviruses are controlled through insecticide control of their mosquito vector. However, inconsiderate use of insecticides often results in the selection of resistance in treated populations, so that monitoring is required to optimize their usage. Here, Culex pipiens (West Nile and Rift Valley Fever virus vector) specimens were collected from four Moroccan cities. Levels of susceptibility to the organophosphate (OP) insecticide malathion were assessed using World Health Organization (WHO)‐recommended bioassays. Individual mosquitoes were tested for the presence of the G119S mutation in the ace‐1 gene, the main OP‐target resistance mutation. RESULTS: Bioassays showed that mosquitoes from Mohammedia were significantly more resistant to malathion than those from Marrakech. Analyzing the ace‐1 genotypes in dead and surviving individuals suggested that other resistance mechanisms may be present in Mohammedia. The ace‐1 resistance allele frequencies were relatively moderate (< 0.4). Their analyses in three Moroccan cities (Tangier, Casablanca and Marrakech) however showed disparities between two coexisting Cx. pipiens forms and revealed that the G119S mutation tends to be more frequent in urban than in rural collection sites. CONCLUSION: These findings provide a reference assessment of OP resistance in Morocco and should help the health authorities to develop informed and sustainable vector control programs. © 2018 Society of Chemical Industry Recurrent vector control selects for insecticide resistance. This article presents an evaluation of target‐mutation resistance to organophosphate insecticides in the Culex pipiens complex in Morocco, and its relation to urbanization. [ABSTRACT FROM AUTHOR]

Anopheles sergentii known as the “oasis vector” or the “desert malaria vector” is considered the main vector of malaria in the southern parts of Morocco. Its presence in Morocco is confirmed for the first time through sequencing of mitochondrial DNA (mDNA) cytochrome c oxidase subunit I (COI) barcodes and nuclear ribosomal DNA (rDNA) second internal transcribed spacer (ITS2) sequences and direct comparison with specimens of A. sergentii of other countries. The DNA barcodes (n = 39) obtained from A. sergentii collected in 2015 and 2016 showed more diversity with 10 haplotypes, compared with 3 haplotypes obtained from ITS2 sequences (n = 59). Moreover, the comparison using the ITS2 sequences showed closer evolutionary relationship between the Moroccan and Egyptian strains than the Iranian strain. Nevertheless, genetic differences due to geographical segregation were also observed. This study provides the first report on the sequence of rDNA-ITS2 and mtDNA COI, which could be used to better understand the biodiversity of A. sergentii. [ABSTRACT FROM AUTHOR]