Entomological Exploration of Sand Flies in Human Communities Affected by Cutaneous and Visceral Leishmaniasis in El Hajeb Province, Morocco.

Sand flies are the exclusive vectors of leishmaniasis. This group of parasitic diseases is a serious public health problem in Morocco. The aim of this study was to investigate the sand fly fauna, mainly the species composition, biodiversity, and seasonal activity of sand flies in El Hajeb in central Morocco. A total of six stations (Aït Naaman, Aït Rbaa, Aït Brahim, Ain Taoujdate, Sidi Mbarek, and Aït Oufella) were studied, five of which had recently recorded cases of leishmaniasis. Sand fly bimonthly captures were carried out using a sticky paper trap in different biotopes from March to December 2019. A total of 14590 adult sand flies were collected. The activity of the sand fly started in April and declined in November. The periods of high abundance were July, September, and November. Morphological identification of sand flies shows the presence of twelve species: Phlebotomus papatasi, P. longicuspis, P. perniciosus, P. sergenti, P. bergeroti, P. alexandri, P. dreyfussi, P. ariasi, Sergentomyia fallax, S. minuta, S. schwetzi, and S. antennata. The analysis showed that species belonging to the genus Phlebotomus were the most dominant (93.3%) and the risk periods were spread during the summer and autumn seasons. The present study provides for the first time information on the species of sand flies in El Hajeb. It, therefore, provides decision makers with an important tool to conduct vector control actions during peak periods in order to limit the transmission of leishmaniasis. A preprint was made available by the research square in the following link: "https://assets.researchsquare.com/files/rs-1409330/v1/dfef7013-0327-4a54-897f-214924a2d950.pdf?c=1646838874."

Ecological Survey of the Peridomestic Sand Flies of an Endemic Focus of Zoonotic Cutaneous Leishmaniasis in the South-East of Morocco.

Leishmaniasis is a parasitosis caused by parasites of the genus Leishmania and is transmitted by Phlebotominae sand flies. An entomological survey was carried out in different localities of Zagora Province. Our work allowed us to establish an inventory of sand flies to study potential vectors of leishmaniasis and to compare the composition and the specific abundance of different endemic stations. The sand flies were collected using CDC miniature light traps during the month of July 2019 in the ten studied villages. The results indicate the presence of thirteen species, belonging to the genera Phlebotomus and Sergentomyia. Phlebotomus papatasi was the predominant species (46.65%) followed by Ph. alexandri (17%), Ph. longicuspis (11.55%), Ph. bergeroti (1.53%) and Ph. sergenti (1.27%). Phlebotomus kazeruni (0.03%) was rare, and only one female was captured in Ifred. Sergentomyia schwetzi (8.69%) was the most prevalent species in the Sergentomyia genus followed closely by Se. fallax (6.84%). Sergentomyia africana was present with a proportion of (3.86%) followed by Se. clydei (1.96%). Sergentomyia dreifussi (0.46%), Se. antennata (0.08%), and Se. minuta (0.08%) were very limited. Phlebotomus papatasi, Ph. alexandri, Ph. bergeroti, Ph. longicuspis, Ph. sergenti, Se. schwetzi, Se. clydei, and Se. fallax are constant species, being present at least in 50% of the stations (occurrence> 50%). Common species (25%-49%) were Se. minuta and Se. africana and rare species were Ph. kazeruni and Se. antennata with a very limited distribution (occurrence <12%). The greatest species richness was found in Ksar Mougni and Ifred with the occurrence of 11 species, but overall, it was high (>9 species) in most of the villages. The Shannon-Wiener index was high (H' > 1) in eight localities (Ksar Mougni, Tassaouante, Bleida, ZaouiteLeftah, Ifred, Timarighine, Ait Oulahyane, and Ait Ali Ouhassou). The high value of this index is in favor of the ZaouiteLeftah locality (Shannon-Wiener index = 1.679) which is explained by the presence of a stand dominated by Ph. papatasi. In order to avoid exposure to infections, a good epidemiological surveillance and vector with rodent control measures must be well maintained. Awareness campaigns are also required and must be conducted for better knowledge of the disease.

Diversity and Bionomics of Sandflies (Diptera: Psychodidae) of an Endemic Focus of Cutaneous Leishmaniasis in Zagora Province, Southeast of Morocco.

The diversity and seasonality for sandflies were studied in 2019 at a focus of zoonotic cutaneous leishmaniasis in Zagora province, southern Morocco. Standardized sampling with CDC light traps was used. A total of 4504 sandflies (4024 Phlebotomus and 480 Sergentomyia) was collected during the study period. Seven species belonging to genus Phlebotomus and six species of genus Sergentomyia were identified. The most abundant species were Ph. papatasi (33.6%) and Ph. longicuspis (25.7%), highlighting the risk for local disease transmission foci. The seasonal activity of sandflies extended from April to November, showing two peaks, one in June-July and one, less important, in late-September-October. Abundance was highest during the months May, June, and July and lowest in August, September, and October. Results of this study provide important baseline data for planning control interventions.

Entomological Investigations, Seasonal Fluctuations and Impact of Bioclimate Factors of Phlebotomines Sand Flies (Diptera: Psychodidae) of an Emerging Focus of Cutaneous Leishmaniasis in Aichoun, Central Morocco.

Leishmaniasis diseases are endemic in Morocco. An entomological survey was conducted in Aichoun locality for 1 year from September 2013 to August 2014. The objective of this study was to investigate the sand fly fauna, mainly the species composition and the monthly species prevalence in accordance with bioclimate factors. Sand flies were collected twice a month, using sticky traps and CDC light traps. During a one-year study, 4472 specimens of sand fly were caught (72.56% male/22.44% female) that were composed of seven species divided into two genera: Phlebotomus (99.46%) and Sergentomyia (0.53%). Ph. sergenti was the most prevalent species (46.64%), followed by Ph. perniciosus (38.19%), Ph. longicuspis (9.32%), Ph. papatasi (5.23%), and Ph. ariasi (0.06%). The genus Sergentomyia was even less frequent. The population dynamics showed a bimodal trend with two peaks: the first one in October (12.03% of specimens) and the second in June (27.92% of specimens). The study of the effects of climatic factors in the study area showed a link between the dynamics of sand flies and the variation of these parameters (temperature and relative humidity). During the period between November and March, the sand flies were absent. The highest prevalence of sand flies was recorded in June when the temperature and relative humidity values reached, respectively, 25.8°C and 42%. The minimum number of specimens was collected in September with an average temperature of 23.19°C and relative humidity of 57.4%. Statistical analysis with principal component analysis has shown a strong positive correlation between temperature parameters and the seasonal distribution of sand flies. The climatic factor of relative humidity has been judged of being negatively correlated. The wind speed does not have any impact on the relative abundance of all species. Within this context, the results will be useful for the development of a monitoring program to better manage the operations and evaluate their effectiveness.

The Current Insecticide Resistance in Main Malaria Vector Anopheles arabiensis in Yemen.

Control of malaria vectors in Yemen relies on both indoor residual spraying using carbamate (bendiocarb) and long-lasting pyrethroids-treated nets. This paper reports the results of studies conducted to monitor the insecticide resistance of the main malaria vector, Anopheles arabiensis, to the insecticides currently used in the vector control in four different locations. Susceptibility tests were performed following the WHO test procedures. Two pyrethroids (lambda-cyhalothrin 0.05% and deltamethrin 0.05%) and one carbamate (bendiocarb 0.1%) were tested at diagnostic doses (DD). The five-fold DD of lambda-cyhalothrin and deltamethrin (0.25%) were also used to yield information on the intensity of resistance. Besides, tests with synergists were performed to assess the involvement of detoxifying enzyme in the phenotypic resistance of the populations of An. arabiensis to pyrethroids. The results of the performed susceptibility bioassay showed that the vector is susceptible to bendiocarb and resistant to lambda-cyhalothrin and deltamethrin in the four studied areas. The pyrethroids resistance is solely metabolic. This information could help policy-makers to plan insecticide resistance management. Bendiocarb is still an effective insecticide in the form of IRS. Concerning LLINS, it would be interesting to assess their effectiveness, combining a pyrethroid with PBO for the control of the pyrethroid-resistant malaria vector.

Towards harmonisation of entomological surveillance in the Mediterranean area.

BACKGROUND: The Mediterranean Basin is historically a hotspot for trade, transport, and migration. As a result, countries surrounding the Mediterranean Sea share common public health threats. Among them are vector-borne diseases, and in particular, mosquito-borne viral diseases are prime candidates as (re)emerging diseases and are likely to spread across the area. Improving preparedness and response capacities to these threats at the regional level is therefore a major issue. The implementation of entomological surveillance is, in particular, of utmost importance. Guidance in designing entomological surveillance systems is critical, and these systems may pursue different specific objectives depending on the disease. The purpose of the proposed review is to draw up guidelines for designing effective and sustainable entomological surveillance systems in order to improve preparedness and response. However, we make it clear that there is no universal surveillance system, so the thinking behind harmonisation is to define evidence-based standards in order to promote best practises, identify the most appropriate surveillance activities, and optimise the use of resources. Such guidance is aimed at policymakers and diverse stakeholders and is intended to be used as a framework for the implementation of entomological surveillance programmes. It will also be useful to collaborate and share information with health professionals involved in other areas of disease surveillance. Medical entomologists and vector control professionals will be able to refer to this report to advocate for tailored entomological surveillance strategies. The main threats targeted in this review are the vectors of dengue virus, chikungunya virus, Zika virus, West Nile virus, and Rift Valley fever virus. The vectors of all these arboviruses are mosquitoes. METHODS: Current knowledge on vector surveillance in the Mediterranean area is reviewed. The analysis was carried out by a collaboration of the medical entomology experts in the region, all of whom belong to the MediLabSecure network, which is currently funded by the European Union and represents an international effort encompassing 19 countries in the Mediterranean and Black Sea region. FINDINGS: Robust surveillance systems are required to address the globalisation of emerging arboviruses. The prevention and management of mosquito-borne viral diseases must be addressed in the prism of a One Health strategy that includes entomological surveillance as an integral part of the policy. Entomological surveillance systems should be designed according to the entomological and epidemiological context and must have well-defined objectives in order to effect a tailored and graduated response. We therefore rely on different scenarios according to different entomological and epidemiological contexts and set out detailed objectives of surveillance. The development of multidisciplinary networks involving both academics and public authorities will provide resources to address these health challenges by promoting good practises in surveillance (identification of surveillance aims, design of surveillance systems, data collection, dissemination of surveillance results, evaluation of surveillance activities) and through the sharing of effective knowledge and information. These networks will also contribute to capacity building and stronger collaborations between sectors at both the local and regional levels. Finally, concrete guidance is offered on the vector of the main arbovirus based on the current situation in the area.

Potential of Aedes albopictus to cause the emergence of arboviruses in Morocco.

In 2015, the mosquito Aedes albopictus was detected in Rabat, Morocco. This invasive species can be involved in the transmission of more than 25 arboviruses. It is known that each combination of mosquito population and virus genotype leads to a specific interaction that can shape the outcome of infection. Testing the vector competence of local mosquitoes is therefore a prerequisite to assess the risks of emergence. A field-collected strain of Ae. albopictus from Morocco was experimentally infected with dengue (DENV), chikungunya (CHIKV), zika (ZIKV) and yellow fever (YFV) viruses. We found that this species can highly transmit CHIKV and to a lesser extent, DENV, ZIKV and YFV. Viruses can be detected in mosquito saliva at day 3 (CHIKV), day 14 (DENV and YFV), and day 21 (ZIKV) post-infection. These results suggest that the local transmission of these four arboviruses by Ae. albopictus newly introduced in Morocco is a likely scenario. Trial registration: ClinicalTrials.gov APAFIS#6573-201606l412077987v2.

Identification of mosquitoes (Diptera: Culicidae): an external quality assessment of medical entomology laboratories in the MediLabSecure Network.

BACKGROUND: Identification of vectors is of prime importance in the field of medical entomology for both operational and research purposes. An external quality assessment of mosquito identification capacities was carried out within the MediLabSecure Network, which is composed of laboratories located in 19 countries close to the European Union around the Mediterranean and Black seas. METHODS: A set of blind samples consisting of 7 or 8 adult mosquitoes and 4 larvae was given to each participant laboratory. In all, 138 adult mosquitoes and 76 larvae of different species were distributed for genus and species identification. RESULTS: All identifications were exclusively morphology based. Overall, 81% of identifications were correct at the genus level, 64% at the species level. The results were highly varied among the 19 participating laboratories. The levels of correct identifications were: 100% (three laboratories), 90-95% (four laboratories), 50-75% (six laboratories) and < 50% (six laboratories). CONCLUSIONS: This evaluation showed the need to maintain efforts in capacity building and quality control in the field of medical entomology and, more specifically, in the morphological identification of the Culicidae.

Insecticide resistance and target site mutations (G119S ace-1 and L1014F kdr) of Culex pipiens in Morocco.

BACKGROUND: Control of the mosquito vector Culex pipiens with insecticides is the main way to control arboviruses that the species can transmit such as West Nile virus (WNV) and Rift Valley fever virus (RVFV). However, its efficiency has been hampered by the emergence of insecticide resistance. Little is known about the insecticide-resistance status and underlying resistance mechanisms of field-collected populations of Cx. pipiens in Morocco. METHODS: Mosquito adults from Mohammadia city in Morocco were reared from immature stages. The level of their susceptibility to insecticides was assessed using standard WHO bioassay. The two forms of the Cx. pipiens complex and their hybrids were identified by a multiplex PCR. Identified mosquitoes were then tested for the presence of the G119S ace-1 and L1014F kdr mutations using PCR-RFLP and PCR assays, respectively. RESULTS: WHO bioassays indicated that Cx. pipiens was resistant to all tested insecticides: lambda-cyhalothrin (49% mortality), permethrin (63% mortality), DDT (16% mortality), malation (52% mortality) and bendiocarb (39% mortality). The frequency of the 119S allele was almost identical in the pipiens form and hybrids (0.11 and 0.15, respectively) whereas it remained low in the molestus form (0.03). No significant correlation was observed between the G119S allele and the resistance phenotype to two tested insecticides (malathion and bendiocarb). The frequency of the L1014F allele was identical in the pipiens form and hybrids (0.44) whereas it was low in the molestus form (0.36) but no significant difference was detected (χ2 = 1.46, df = 1, P = 0.225). The presence of the L1014F kdr mutation was significantly associated with resistance to three tested insecticides in pipiens form (P = 0.0019, P = 0.0023 and P = 0.023, respectively, to lambda-cyhalothrin, permethrin and DDT) whereas no significant correlation was observed between the L1014F kdr mutation and resistance phenotype in molestus form and hybrids to the three tested insecticides. CONCLUSION: These findings showed that wild populations of Cx. pipiens have developed resistance against the main insecticide families with different modes of action: organochlorines (DDT), organophosphates (malathion), carbamates (bendiocarb), pyrethroids (lambda-cyhalothrin, permethrin). Therefore, urgent action should be taken to manage the resistance in this species to maintain the effectiveness of arbovirus control.

Surveillance of Arthropod-Borne Viruses and Their Vectors in the Mediterranean and Black Sea Regions Within the MediLabSecure Network.

PURPOSE OF REVIEW: Arboviruses, viruses transmitted by arthropods such as mosquitoes, ticks, sandflies, and fleas are a significant threat to public health because of their epidemic and zoonotic potential. The geographical distribution of mosquito-borne diseases such as West Nile (WN), Rift Valley fever (RVF), Dengue, Chikungunya, and Zika has expanded over the last decades. Countries of the Mediterranean and Black Sea regions are not spared. Outbreaks of WN are repeatedly reported in the Mediterranean basin. Human cases of RVF were reported at the southern borders of the Maghreb region. For this reason, establishing the basis for the research to understand the potential for the future emergence of these and other arboviruses and their expansion into new geographic areas became a public health priority. In this context, the European network "MediLabSecure" gathering laboratories in 19 non-EU countries from the Mediterranean and Black Sea regions seeks to improve the surveillance (of animals, humans, and vectors) by reinforcing capacity building and harmonizing national surveillance systems to address this important human and veterinary health issue. The aim of this review is to give an exhaustive overview of arboviruses and their vectors in the region. RECENT FINDINGS: The data presented underline the importance of surveillance in the implementation of more adapted control strategies to combat vector-borne diseases. Partner laboratories within the MediLabSecure network present a wide range of infrastructures and have benefited from different training programs. SUMMARY: Although reporting of arboviral presence is not carried out in a systematic manner, the expansion of the area where arboviruses are present cannot be disputed. This reinforces the need for increasing surveillance capacity building in this region to prevent future emergences.

First report of L1014F-kdr mutation in Culex pipiens complex from Morocco.

BACKGROUND: Mosquitoes of the Culex pipiens complex, competent vectors for West Nile virus (WNV) and Rift Valley fever virus (RVFV) are widely targeted by insecticide treatments. The intensive application of chemical insecticides led to the development of resistance in many insects including Culex pipiens mosquitoes. The absence of data on resistance mechanisms in Morocco allow us to assess the levels of lambda-cyhalothrin resistance and the frequency of the mutated gene L1014F kdr in different forms of Cx. pipiens complex from three regions of Morocco. METHODS: Mosquito adults were reared from immature stages collected in three different regions in Morocco (Tangier, Casablanca and Marrakech). Standard WHO insecticide susceptibility tests were conducted on adults emerged from collected larvae. Specimens were identified as belonging to the Culex pipiens complex using a multiplex PCR assay with diagnostic primers designed from the flanking region of microsatellite CQ11. Identified mosquitoes were then tested for the presence of the L1014F kdr mutation using PCR assay. RESULTS: Our results showed that 21% of the tested population has a resistance to lambda-cyhalothrin. The molecular identification of survivors shows that 43% belonged to the Cx. pipiens pipiens and only 9.5% to the Cx. pipiens molestus form. On the other hand, 416 specimens were screened for the L1014F kdr mutation. L1014F mutation was detected in different forms of Cx. pipiens in different sites. The frequency of L1014F mutation was similar between the Cx. pipiens pipiens form and hybrid form, while it was lower in the Cx. pipiens molestus form. The presence of the L1014F kdr allele was significantly associated with resistance to lambda-cyhalothrin in Cx. pipiens pipiens (P < 0.0001) and hybrid form (P < 0.0001). CONCLUSION: Resistance to lambda-cyhalothrin of Cx. pipiens populations appears to be largely due to the L1014F kdr mutation. To our knowledge, the frequencies of L1014F kdr mutation are examined for the first time in natural populations of the Culex pipiens complex in Morocco. These findings will provide important information to propose more adapted vector control measures towards this mosquito species, potential vector of arboviruses.

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.

Leishmaniasis in central morocco: seasonal fluctuations of phlebotomine sand fly in aichoun locality, from sefrou province.

Cutaneous leishmaniases (CL) are endemic in Morocco. They are common in the human population in different localities such as Aichoun in Sefrou province, Morocco. This study was carried out in Aichoun locality from April to October 2012 in order to study the spatiotemporal trends of the main Leishmania phlebotomine vectors in this focus. Overall, 1171 sand flies, belonging to four species, were collected by sticky traps. Phlebotomus sergenti was the predominant species (78.4%) followed by Ph. perniciosus (10.5%), Ph. papatasi (7.94%), and Ph. longicuspis (3.16%). Sandflies were active during 6 months (May-October). Ph. sergenti, Ph. perniciosus, and Ph. papatasi displayed a bimodal distribution with a first peak in July and a second peak in September, while Ph. longicuspis showed a monophasic trend with a peak in August. The high abundance and the lengthy period of activity of Ph. sergenti and Ph. perniciosus, vectors of L. tropica and L. infantum, respectively, are a cause for concern as they indicate the high potential risk of Leishmania transmission in the studied areas.

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.

Ecological and control techniques for sand flies (Diptera: Psychodidae) associated with rodent reservoirs of leishmaniasis.

BACKGROUND: Leishmaniasis remains a global health problem because of the substantial holes that remain in our understanding of sand fly ecology and the failure of traditional vector control methods. The specific larval food source is unknown for all but a few sand fly species, and this is particularly true for the vectors of Leishmania parasites. We provide methods and materials that could be used to understand, and ultimately break, the transmission cycle of zoonotic cutaneous leishmaniasis. METHODS AND FINDINGS: We demonstrated in laboratory studies that analysis of the stable carbon and nitrogen isotopes found naturally in plant and animal tissues was highly effective for linking adult sand flies with their larval diet, without having to locate or capture the sand fly larvae themselves. In a field trial, we also demonstrated using this technique that half of captured adult sand flies had fed as larvae on rodent feces. Through the identification of rodent feces as a sand fly larval habitat, we now know that rodent baits containing insecticides that have been shown in previous studies to pass into the rodents' feces and kill sand fly larvae also could play a future role in sand fly control. In a second study we showed that rubidium incorporated into rodent baits could be used to demonstrate the level of bloodfeeding by sand flies on baited rodents, and that the elimination of sand flies that feed on rodents can be achieved using baits containing an insecticide that circulates in the blood of baited rodents. CONCLUSIONS: Combined, the techniques described could help to identify larval food sources of other important vectors of the protozoa that cause visceral or dermal leishmaniasis. Unveiling aspects of the life cycles of sand flies that could be targeted with insecticides would guide future sand fly control programs for prevention of leishmaniasis.

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.

DNA barcodes confirm the presence of a single member of the Anopheles maculipennis group in Morocco and Algeria: An. sicaulti is conspecific with An. labranchiae.

Anopheles labranchiae Falleroni is the only member of the Maculipennis Group known to occur in northern Africa; however, confusion exists as to the taxonomic status of its junior synonym, An. sicaulti Roubaud (type locality: near Rabat, Morocco). Based on morphological and behavioural distinctions, it has been suggested that Moroccan populations have been isolated from other North African populations by the Atlas Mountains, and that Moroccan populations may represent An. sicaulti, originally described as a variety of An. maculipennis Meigen. DNA barcodes (658bp of the mitochondrial COI gene) obtained from 89 An. maculipennis s.l. collected in Morocco (n=79) and Algeria (n=10) in 2007 and 2008 were used to determine if Moroccan populations are genetically isolated from those east of the Atlas Mountains (Algeria), and whether there is molecular evidence to support the presence of more than one member of the Maculipennis Group in the region. No evidence for speciation was found between Moroccan and Algerian populations, or within populations in northern Morocco. Moreover shared COI haplotypes between Algeria and Morocco indicate ongoing gene flow between populations in these countries, suggesting that the Atlas Mountains are not a boundary to gene flow in An. labranchiae. The synonymy of An. sicaulti with An. labranchiae is confirmed. That An. labranchiae comprises the same species in these North African countries is important for malaria control.