Composition and diversity of Culicoides biting midges (Diptera: Ceratopogonidae) in rural and suburban environments of Algeria.

Culicoides biting midges are well known biological vectors of several arboviruses causing more than 100 veterinary and medical diseases worldwide. In Algeria, bluetongue virus, which is transmitted by Culicoides midges, is responsible for one of the most critical insect-borne diseases of sheep. For example, this disease caused clinically severe morbidity of about 2,661 confirmed cases out of 21,175 susceptible sheep in Algeria. This study compared the abundance, richness, and diversity of Culicoides species in rural and suburban environments of semi-arid regions in North Africa. It examined the potential influence of the bio-climatic factors on the variation of population sizes and the elevation distribution of biting midges. Specimen collection was carried out from June to September during two successive years (2018 and 2019) using CDC light traps installed at 15 sites in different environments. Culicoides specimens were dissected, slide-mounted, and morphologically identified using the interactive identification key IIKC and various standard morphological criteria. A total of 1,046 Culicoides specimens (1,024 females and 22 males) were trapped and classified into 22 species, belonged to nine distinct subgenera. Two new species records for Algeria and even North Africa are reported: Culicoides albicans (Winnertz, 1852) and Culicoides nubeculosus (Meigen, 1830). Culicoides newsteadi Austen, 1921 (51.6%) was the dominant species, and it was followed by Culicoides punctatus (Meigen, 1804) (16.3%) and Culicoides odiatus Austen, 1921 (11.5%). These three species, comprising 80% of the collected Culicoides, were the most abundant both outside and inside livestock stables in rural and suburban environments. Species diversity was similar in the two settings, with a slight increase in suburban environments. None of the Culicoides species encountered correlated significantly with the climatic factors (mean temperature, precipitation, and relative humidity). Elevation was the most determinant environmental parameter that affected the abundance and distribution of Culicoides midges in the semi-arid and sub-humid areas studied. The maximum distribution of Culicoides species was detected at mid elevations (400‒800 m). Using a modeling approach, we explored for the first time the variation of composition and diversity in Culicoides communities within different climatic regions, environments and livestock settings in Algeria. This survey deepens our understanding of the relationships among environmental factors, abundance, diversity, and geographic distribution of Culicoides. This is a crucial step to assess the epidemiological situation of the diseases transmitted by these biting midges and to allow mitigation of the associated risks.

Sand Flies (Diptera: Psychodidae): Fauna and Ecology in the Northeast of Algeria.

Sand flies (Diptera: Psychodidae) transmit several Leishmania (Kinetoplastida: Trypanosomatidae) species, which cause leishmaniasis, a significant public health concern in Algeria. We compared sand fly species abundance and composition among different biotopes (urban, peri-urban, rural areas), bioclimatic zones (humid, sub-humid, semi-arid), and elevation ranges. We also used the additive partitioning of beta diversity to test whether the variation in sand fly composition among biotopes, bioclimatic zones, and elevation ranges is due to species turnover or community subsetting. In total, 7,478 specimens were captured; of which, 7,162 (51.5% males vs. 48.5% females) belong to eight species: Phlebotomus perniciosus Newstead, 1911 (77.4% of the total captured specimens), Phlebotomus perfiliewi Parrot, 1930 (14.6%), Phlebotomus longicuspis Nitzulescu, 1911 (5.9%), Phlebotomus papatasi Scopoli, 1786 (<1%), Phlebotomus sergenti Parrot, 1917 (<1%) and Phlebotomus chadlii Rioux, Juminer et Gibily 1966 (<1%), Sergentomyia minuta Adler et Theodor, 1927 (1%), and Sergentomyia fallax Parrot, 1921 (<1%). Sand fly total abundance showed negative correlations with altitude and was significantly higher in rural areas. Sandfly community composition was significantly different between rural and urban/peri-urban areas. The additive partitioning of beta diversity showed that 71.4% of the compositional dissimilarity among elevation ranges and bioclimates was due to sand fly species turnover, and 28.6% resulted from nestedness. However, the variation in sand fly composition among different biotopes was mainly due to community nestedness. Findings from this study help define the risk of Leishmania transmission and develop methods for vector control in Mila province and Algeria as a whole.

Limits of a rapid identification of common Mediterranean sandflies using polymerase chain reaction-restriction fragment length polymorphism

A total of 131 phlebotomine Algerian sandflies have been processed in the present study. They belong to the species Phlebotomus bergeroti, Phlebotomus alexandri, Phlebotomus sergenti, Phlebotomus chabaudi, Phlebotomus riouxi, Phlebotomus perniciosus, Phlebotomus longicuspis, Phlebotomus perfiliewi, Phlebotomus ariasi, Phlebotomus chadlii, Sergentomyia fallax, Sergentomyia minuta, Sergentomyia antennata, Sergentomyia schwetzi, Sergentomyia clydei, Sergentomyia christophersi and Grassomyia dreyfussi. They have been characterised by sequencing of a part of the cytochrome b (cyt b), t RNA serine and NADH1 on the one hand and of the cytochrome C oxidase I of the mitochondrial DNA (mtDNA) on the other hand. Our study highlights two sympatric populations within P. sergenti in the area of its type-locality and new haplotypes of P. perniciosus and P. longicuspis without recording the specimens called lcx previously found in North Africa. We tried to use a polymerase chain reaction-restriction fragment length polymorphism method based on a combined double digestion of each marker. These method is not interesting to identify sandflies all over the Mediterranean Basin.

Limits of a rapid identification of common Mediterranean sandflies using polymerase chain reaction-restriction fragment length polymorphism.

A total of 131 phlebotomine Algerian sandflies have been processed in the present study. They belong to the species Phlebotomus bergeroti, Phlebotomus alexandri, Phlebotomus sergenti, Phlebotomus chabaudi, Phlebotomus riouxi, Phlebotomus perniciosus, Phlebotomus longicuspis, Phlebotomus perfiliewi, Phlebotomus ariasi, Phlebotomus chadlii, Sergentomyia fallax, Sergentomyia minuta, Sergentomyia antennata, Sergentomyia schwetzi, Sergentomyia clydei, Sergentomyia christophersi and Grassomyia dreyfussi. They have been characterised by sequencing of a part of the cytochrome b (cyt b), t RNA serine and NADH1 on the one hand and of the cytochrome C oxidase I of the mitochondrial DNA (mtDNA) on the other hand. Our study highlights two sympatric populations within P. sergenti in the area of its type-locality and new haplotypes of P. perniciosus and P. longicuspis without recording the specimens called lcx previously found in North Africa. We tried to use a polymerase chain reaction-restriction fragment length polymorphism method based on a combined double digestion of each marker. These method is not interesting to identify sandflies all over the Mediterranean Basin.

A taxonomic study of Phlebotomus (Larroussius) perfiliewi s. l.

Phlebotomus (Larroussius) perfiliewi is one of the main vectors of Leishmania infantum in the Mediterranean basin and in Central Asia. Its taxonomic status remains doubtful. It usually includes three taxa of specific or subspecific value, depending on the authors: P. perfiliewiParrot, 1930 s. st. in North Africa, Western Europe and the Balkans, Crimea; Phlebotomus galilaeusTheodor, 1958 in Israel/Palestine and Cyprus and Phlebotomus transcaucasicusPerfiliev, 1937 in Caucasus and Central Asia. While the identification of the females of P. perfiliewi s. l. is relatively easy, based on the particular morphology of their spermathecal ducts, the specific identification of the females of Phlebotomus perfiliewi perfiliewi, Phlebotomus perfiliewi galilaeus and Phlebotomus perfiliewi transcaucasicus is impossible. The differentiation of males, based mainly on characters on aedeagus, is also difficult. The goal of the present study is to carry out a taxonomic update based on a morphological study of specimens coming from various populations from Asia, Europe and North Africa coupled with their molecular analysis.

Morphological and molecular investigations of population structure of Phlebotomus perniciosus and Phlebotomus longicuspis (Diptera: Psychodidae) in Tunisia.

Morphological and molecular characterization of Phlebotomus (Larroussius) perniciosus Newstead and Phlebotomus (L.) longicuspis Nitzulescu in Tunisia is reported. Different localities in central and southern Tunisia were sampled. Sand flies were collected by sticky-paper traps and Center for Disease Control traps. For morphological study of males, the copulatory valves (aedeagi) were examined and the number of coxite hairs was recorded. For molecular analysis, the mitochondrial cytochrome b gene and the cytochrome c oxidase I gene were sequenced to investigate the population structure of P. perniciosus and P. longicuspis. The majority of P. perniciosus samples from southern and some specimens from central Tunisia showed single-pointed aedeagi curved at their apicies that were indistinguishable from the P. longicuspis aedeagi and appeared similar to the atypical morph of P. perniciosus, previously described in northern Morocco. The current study gives evidence of a wide distribution of atypical morphs of P. perniciosus in southern and central Tunisia.

Genetic structure of Phlebotomus (Larroussius) ariasi populations, the vector of Leishmania infantum in the western Mediterranean: epidemiological implications.

In recent years there has been growing interest in analyzing the geographical variations between populations of different Phlebotomus spp. by comparing the sequences of various genes. However, little is known about the genetic structure of Phlebotomus ariasi. In this study, we were able to sequence a fragment of the mitochondrial Cyt b gene in 133 sandflies morphologically identified as P. ariasi and proceeding from a wide geographical range covering 35 locations in 11 different regions from five countries. The intra-specific diversity of P. ariasi is high, with 45 haplotypes differing from each other by one to 26 bases and they are distributed in two mitochondrial lineages, one limited geographically to Algeria and the other widely dispersed across Mediterranean countries. The Algerian lineage is characterized by having 13 fixed polymorphisms and is made up of one sole haplotype. The European/Moroccan P. ariasi lineage is characterized by being made up of a great diversity of haplotypes (44) which display some geographical structuring. This could be one of the multiple factors involved in the epidemiological heterogeneity of the foci of leishmaniasis. Phlebotomus chadlii is the sister group of European/Moroccan P. ariasi. The separation of the Algerian haplotype, H45, from the rest of the specimens, European/Moroccan P. ariasi and P. chadlii, is well supported by the bootstrap analysis.

Molecular homogeneity in diverse geographical populations of Phlebotomus papatasi (Diptera, Psychodidae) inferred from ND4 mtDNA and ITS2 rDNA Epidemiological consequences.

An intraspecific study on Phlebotomus papatasi, the main proven vector of Leishmania major among the members of the subgenus Phlebotomus, was performed. The internal transcribed spacer 2 (ITS 2) of rDNA and the ND4 gene of mt DNA were sequenced from 26 populations from 18 countries (Albania, Algeria, Cyprus, Egypt, Greece, India, Iran, Israel, Italy, Lebanon, Morocco, Saudi Arabia, Spain, Syria, Tunisia, Turkey, Yugoslavia and Yemen), and compared. Samples also included three other species belonging to the subgenus Phlebotomus: P. duboscqi, a proven vector of L. major in the south of Sahara (three populations from Burkina Faso, Kenya and Senegal), P. bergeroti, a suspected vector of L. major (three populations from Oman Sultanate, Iran and Egypt), and one population of P. salehi from Iran. A phylogenetic study was carried out on the subgenus Phlebotomus. Our results confirm the validity of the morphologically characterized taxa. The position of P. salehi is doubtful. Variability in P. papatasi contrasts with that observed within other species having a wide distribution like P. (Paraphlebotomus) sergenti in the Old World or Lutzomyia (Lutzomyia) longipalpis in the New World. Consequently, it could be hypothesized that all populations of P. papatasi over its distribution area have similar vectorial capacities. The limits of the distribution area of L. major are correlated with the distribution of common rodents acting as hosts of the parasites.