Malpighian tubules are important determinants of Pseudomonas transstadial transmission and longtime persistence in Anopheles stephensi

Background: Pseudomonas is a genus of bacteria commonly found in investigations of gut microbes in malaria mosquitoes. Among those mosquitoes is the dominating malaria vector in Asia, Anopheles stephensi, where Pseudomonas is a prevailing bacterium and natural inhabitant of its breeding places. In order to explore the reason for finding Pseudomonas so frequently, an investigation of its localization and transstadial properties was undertaken.Methods: A Pseudomonas isolate from Anopheles stephensi was transformed successfully with an endogenous plasmid modified to express green fluorescent protein (GFP). Subsequently, the Pseudomonas-GFP was added to the laboratory larval breeding place of Anopheles stephensi and taken up by the larvae. After 24 hours, the larvae were cleaned and moved to a bath with double-distilled water. Also, female adults were fed sugar solution containing Pseudomonas-GFP. The Pseudomonas-GFP was traced in the alimentary canal of larvae, pupae and adults.Results: Fluorescent microscopy and PCR assays showed that the Pseudomonas bacteria underwent transstadial transmission from larvae to pupae and then to adults. In blood-fed female mosquitoes, the bacteria increased in numbers and remained in the mosquito body for at least three weeks after eclosion. In addition to the midgut, the Malpighian tubules of both larvae and adult mosquitoes were colonized by the bacteria. Also Pseudomonas-GFP that was distributed through sugar solution was able to colonize the Malpighian tubules of adult females.Conclusions: Colonization of the Malpighian tubules by Pseudomonas bacteria seems to be important for the transstadial passage from larvae to adult and presumably for the longevity of the bacteria in the adult mosquito. The existence of an entry point in the larval stage, and the long duration in the female gut, opens up for a possible use of Pseudomonas in mosquito paratransgenesis

Molecular analysis of the mitochondrial markers COI, 12S rDNA and 16S rDNA for six species of Iranian scorpions

Objectives Annually, 1.2 million humans are stung by scorpions and severely affected by their venom. Some of the scorpion species of medical importance have a similar morphology to species with low toxicity. To establish diagnostic tools for surveying scorpions, the current study was conducted to generate three mitochondrial markers, Cytochrome Oxidase I (COI gene), 12S rDNA and 16S rDNA for six species of medically important Iranian scorpions: Androctonus crassicauda, Hottentotta saulcyi, Mesobuthus caucasicus, M. eupeus, Odontobuthus doriae, and Scorpio maurus. Results Phylogenetic analyses of the obtained sequences corroborated the morphological identification. For the first time, 12S rDNA sequences are reported from Androctonus crassicauda, Hottentotta saulcyi, Mesobuthus caucasicus and M. eupeus and also the 16S rDNA sequence from Hottentotta saulcyi. We conclude that the mitochondrial markers are useful for species determination among these medically important species of scorpions

Mapping local patterns of childhood overweight and wasting in low- and middle-income countries between 2000 and 2017

A double burden of malnutrition occurs when individuals, household members or communities experience both undernutrition and overweight. Here, we show geospatial estimates of overweight and wasting prevalence among children under 5 years of age in 105 low- and middle-income countries (LMICs) from 2000 to 2017 and aggregate these to policy-relevant administrative units. Wasting decreased overall across LMICs between 2000 and 2017, from 8.4% (62.3 (55.1–70.8) million) to 6.4% (58.3 (47.6–70.7) million), but is predicted to remain above the World Health Organization’s Global Nutrition Target of <5% in over half of LMICs by 2025. Prevalence of overweight increased from 5.2% (30 (22.8–38.5) million) in 2000 to 6.0% (55.5 (44.8–67.9) million) children aged under 5 years in 2017. Areas most affected by double burden of malnutrition were located in Indonesia, Thailand, southeastern China, Botswana, Cameroon and central Nigeria. Our estimates provide a new perspective to researchers, policy makers and public health agencies in their efforts to address this global childhood syndemic

Larval habitats and species diversity of mosquitoes (Diptera: Culicidae) in West Azerbaijan Province, Northwestern Iran

Abstract Background The characteristics of a larval habitat is an important factor which affects the breeding pattern and population growth of mosquitoes Information about the larval habitat characteristics and pupal productivity can be utilized for the surveillance of the level of population growth, species diversity, and preferred breeding sites of mosquitoes, which are important aspects of integrated vector control. In the present study, mosquito larvae were collected from 22 natural habitats in five counties of the West Azerbaijan Province in the Northwest of Iran during May–November 2018. Physicochemical characteristics of the habitats were investigated. These included alkalinity, chloride (Cl) content, water temperature (°C), turbidity (NTU), Total Dissolved Solids (TDS) (ppm), Electrical Conductivity (EC) (μS/cm), and acidity (pH). The index of affinity between the collected species was calculated using Fager & McGowan test. Results A total of 2715 specimens were collected and identified. Seven different species belonging to four genera were identified in our study sites. The species included, Culex pipiens Linnaeus 1758, Culex theileri Theobald 1903, Culex mimeticus Noé 1899, Culex modestus Ficalbi 1947, Culiseta longiareolata Macquart 1838, Anopheles maculipennis Meigen 1818complex, and Aedes caspius Pallas 1771. There was a significant difference in chloride content and water temperature preferences among the different species (P  0.05). The affinity between the pair of species Cx. mimeticus/Cs. longiareolata was 0.526. There was no affinity between other pairs of species or the affinity was very weak. Conclusions The physicochemical and biological characteristics of mosquito larval habitats play an important role in zoning of areas suitable for breeding and distribution. Surveillance of these characteristics can provide valuable information for entomological monitoring of mosquito vectors and for designing targeted control programs. Also, further studies should be undertaken in a wider geographical area, taking into account the complex characteristics of the physicochemical and ecological factors of the study area and their interaction with various mosquito species

Susceptibility of Anopheles maculipennis to different classes of insecticides in West Azarbaijan Province, Northwestern Iran

Objective: To determine the susceptibility status of Anopheles maculipennis (An. maculipennis) against the major insecticides used in the health sectors in West Azarbaijan Province, Northwestern Iran. Methods: Unfed 3-5 days old adult females of An. maculipennis were collected across the West Azarbaijan Province and were subjected to evaluation of their susceptibility following World Health Organization recommended protocol against six insecticides (permethrin, deltamethrin, propoxur, bendiocarb, malathion and dieldrin) belonging to four different classes. Results: In this study, 916 specimens of An. maculipennis were examined against the insecticides which indicated that An. maculipennis was tolerant to permethrin, deltamethrin and dielderin, but displayed resistance against propoxur, bendiocarb and malathion. Conclusions: The pattern of resistance in An. maculipennis could be attributed to the agricultural landscapes, agricultural pesticides used and the exposure of the mosquitoes to insecticides. Logical cooperation is needed between the agriculture and health sectors to ensure the judicious use of pesticides in each sector and the management of probable resistance

Molecular characterization, biological forms and sporozoite rate of Anopheles stephensi in southern Iran

Objective: To identify the biological forms, sporozoite rate and molecular characterization of the Anopheles stephensi (An. stephensi) in Hormozgan and Sistan-Baluchistan provinces, the most important malarious areas in Iran. Methods: Wild live An. stephensi samples were collected from different malarious areas in southern Iran. The biological forms were identified based on number of egg-ridges. Molecular characterization of biological forms was verified by analysis of the mitochondrial cytochrome oxidase subunit I and II (mtDNA-COI/COII). The Plasmodium infection was examined in the wild female specimens by species-specific nested–PCR method. Results: Results showed that all three biological forms including mysorensis, intermediate and type are present in the study areas. Molecular investigations revealed no genetic variation between mtDNA COI/COII sequences of the biological forms and no Plasmodium parasites was detected in the collected mosquito samples. Conclusions: Presence of three biological forms with identical sequences showed that the known biological forms belong to a single taxon and the various vectorial capacities reported for these forms are more likely corresponded to other epidemiological factors than to the morphotype of the populations. Lack of malaria parasite infection in An. stephensi, the most important vector of malaria, may be partly due to the success and achievement of ongoing active malaria control program in the region

Phylogenetic analysis of the oriental-Palearctic-Afrotropical members of Anopheles (Culicidae: Diptera) based on nuclear rDNA and mitochondrial DNA characteristics

The phylogenetic relationships of Anopheles spp. at the junction of Oriental, Palearctic, and Afrotropical regions in the Iranian plateau were investigated using molecular markers. A 711-bp mitochondrial DNA cytochrome oxidase C subunit I (COI) fragment and the entire second internal transcribed spacer (ITS2) region (286-576 bp) of the nuclear ribosomal DNA (rDNA-ITS2) were sequenced from 14 and 28 taxa, respectively. The analyses included 12 species within Anopheles and 4 within the Myzorhynchus Series of the subgenus Anopheles, 8 within Neocellia, 6 within Myzomyia, 3 within Paramyzomyia, and 1 within the Pyretophorus Series of the subgenus Cellia. The congruent tree topologies of both molecular markers strongly supported monophyly of subgenera Anopheles and Cellia. Phylogenetic trees constructed on the basis of ITS2 sequences could accurately categorize all of the series according to the classical taxonomy but could not distinguish Pyretophorus (Anopheles subpictus) from Paramyzomyia Series. Although sequence data of the COI region were available for only 14 species, the inferred trees revealed good classification among the series but could not show the monophyletic relationship of Cellia spp. Except for a few cases, the tree inferred from ITS2 sequences revealed the best classification for the species studied. The molecular data could significantly improve our understanding of the phylogenetic position of the taxa

Fauna and Larval Habitats of Mosquitoes (Diptera: Culicidae) of West Azerbaijan Province, Northwestern Iran.

Background: Several important diseases are transmitted by mosquitoes. Despite of the potential of the occurrence of some mosquito-borne diseases such as West Nile, dirofilariasis and malaria in the region, there is no recent study of mosquitoes in West Azerbaijan Province. The aim of this investigation was to study the fauna, composition and dis- tribution of mosquitoes and the characteristics of their larval habitats in this province. Methods: Larvae and adult collections were carried out from different habitats using the standard methods in twenty five localities of seven counties across West Azerbaijan Province. Results: Overall, 1569 mosquitoes including 1336 larvae and 233 adults were collected from 25 localities. The de- tails of geographical properties were recorded. Five genera along with 12 species were collected and identified in- cluding: Anopheles claviger, An. maculipennis s.l., An. superpictus, Culex pipiens, Cx. theileri, Cx. modestus, Cx. hortensis, Cx. mimeticus, Culiseta Longiareolata, Ochlerotatus caspius s.l., Oc. geniculatus   and Uranotaenia unguiculata. This is the first record of Oc. geniculatus in the province. Conclusion: Due to the geographical location of the West Azerbaijan Province, it comprises different climatic con- dition which provides suitable environment for the establishment of various species of mosquitoes. The solidarity geographical, cultural and territorial exchanges complicate the situation of the province and its vectors as a threat for future and probable epidemics of mosquito-borne diseases