Urban landscape as a determinant of sarcosaprophagous fly assemblages of sanitary interest: a comparative field study between larviparous and oviparous reproductive strategies.

In Diptera, reproductive strategies vary according to the stage of development of the newly deposited offspring and their abundance. The aim of our study is to establish connections between the reproductive strategies of flies (larviparous or oviparous) and the prevailing local conditions (sun or shade) as well as landscape attributes (low or high urbanization) in an urban setting. We collected flies using 2 baited traps (in the sun or shade) at each of the 13 study sites with varying levels of urbanization. We used generalized linear mixed models to examine the impact of landscape and local characteristics on the diversity (abundance and richness of larviparous and oviparous Diptera) of sarcosaprophagous flies. Our findings indicate that while the abundance and diversity of both larviparous and oviparous flies are affected by urbanization, larviparous flies would be less sensitive to more urbanized environments. Larviparous also exhibit a preference for resources under sun exposure, while the responses of oviparous seem more species-dependent. The observed patterns can be explained by the known biology of the studied groups.

Synthesis, characterization and larvicidal studies of ethyl 3-benzoyl-7-(piperidin-1-yl)indolizine-1-carboxylate analogues against Anopheles arabiensis and cheminformatics approaches.

According to WHO, in 2021, there was an estimation of 247 million malaria cases from 84 malaria-endemic countries. Globally an estimated count of 2 billion malaria cases and 11.7 million deaths due to malaria were recorded in the past two decades. Further, the emergence of drug-resistant mosquitos threatens mankind. Therefore, the development of newer larvicidal agents is the need of the hour. This research identifies a new series of variably substituted indolizines for their effectiveness in controlling Anopheles arabiensis larvae through larvicidal activity. The series of Ethyl 3-benzoyl-7-(piperidin-1-yl)indolizine-1-carboxylate analogues (4a-j) were synthesized by reacting 4-(piperidin-1-yl)pyridine, phenacyl bromides with ethyl propiolate via 1, 3-dipolar cycloaddition and the green metrics of the process are reported. All the newly synthesized compounds were characterized by spectroscopic techniques such as 1H NMR,13C NMR, FT-IR, and HRMS. The larvicidal effectiveness of the newly synthesized compounds was assessed against Anopheles arabiensis. Among the compounds studied, namely 4c, 4d, 4e, and 4f, displayed the most notable larval mortality rates within the series, reaching 73%, 81%, 76%, and 71% respectively, in contrast with the negative control acetone. In comparison, the standard Temephos exhibited a mortality rate of 99% at the same concentration. Furthermore, computational approaches including molecular docking and molecular dynamics simulations identified the potential targets of the series compounds as the larval Acetylcholinesterase (AChE) enzyme and the Sterol Carrier Protein-2 (SCP-2) protein. However, it is essential for these computational predictions to undergo experimental validation.Communicated by Ramaswamy H. Sarma.

Aedes aegypti oviposition dynamics in towns with low human population density in yungas and dry chaco, Salta, Argentina.

We assessed the presence of Aedes aegypti in five ecorregions of Salta province and compared the oviposition activity of Ae. aegypti using ovitraps in towns of two contrasting ecoregions (yungas and Chaco dry forests) in the province of Salta, Argentina, a major contrast in these ecoregions are rain patterns and altitude. Our aim was to estimate how oviposition activities were associated with the ecoregion and site scale local environmental variables. Mosquito oviposition activity was monitored weekly during the summer using ovitraps. Predictor variables were ecoregion, town, and meteorological variables. The effect of the predictor variables was measured on the response variables using multi-model inference. Besides yungas, the presence of Aedes aegypti was confirmed in towns of dry Chaco and High Monte. The only factor that had a significant effect on the presence of eggs in the ovitraps was the ecoregion, with the frequency of positives being higher in yungas. For the number of eggs, the ecoregion, the night temperature of the first week and the NDVI would explain said variable. Overall, results indicate that the variations between towns would be more related with their ecological and climatic characteristics than with the more immediate meteorological variations.

Synthesis, biological evaluation, and computational investigation of ethyl 2,4,6-trisubstituted-1,4-dihydropyrimidine-5-carboxylates as potential larvicidal agents against Anopheles arabiensis.

Malaria is one of the most known vector-borne diseases caused by female Anopheles mosquito bites. According to WHO, about 247 million cases of malaria and 619,000 deaths were estimated worldwide in 2021, of which 95% of the cases and 96% of deaths occurred in the African region. Sadly, about 80% of all malaria deaths were of children under five years old. Despite the availability of different insecticides used to control this disease, the emergence of drug-resistant mosquitoes threatens public health. This, in turn, highlighted the need for new larvicidal agents that are effective at different larval life stages. This study aimed to identify novel larvicidal agents. To this end, a series of ethyl 2,4,6-trisubstituted-1,4-dihydropyrimidine-5-carboxylates 8a-i was synthesized using a three-step chemical synthetic approach via a Biginelli reaction employed as a key step. All title compounds were screened against Anopheles arabiensis to determine their larvicidal activities. Among them, two derivatives, ethyl 2-((4-bromophenyl)amino)-4-(4-fluorophenyl)-6-methyl-1,4-dihydropyrimidine-5-carboxylate 8b and ethyl 2-((4-bromo-2-cyanophenyl)amino)-4-(4-fluorophenyl)-6-methyl-1,4-dihydropyrimidine-5-carboxylate 8f, showed the highest larvicidal activity, with mortality of 94% and 91%, respectively, and emerged as potential larvicidal agents. In addition, computational studies, including molecular docking and molecular dynamics simulations, were carried out to investigate their mechanism of action. The computational results showed that acetylcholinesterase appears to be a plausible molecular target for their larvicidal property.Communicated by Ramaswamy H. Sarma.

Damselflies (Coenagrionidae) have been avoiding leaf veins during oviposition for at least 52 million years.

Plant-insect interactions can provide extremely valuable information for reconstructing the oviposition behavior. We have studied about 1350 endophytic egg traces of coenagrionid damselflies (Odonata: Zygoptera) from the Eocene, identifying triangular or drop-shaped scars associated with them. This study aims to determine the origin of these scars. Our behavioral study of about 1,800 endophytic eggs from recent coenagrionids indicates that these scars were caused by ovipositor incisions, but without egg insertion. The scar correlates (χ2-test) with leaf veins in both fossil and extant species. We infer that a female would detect the proximity of a leaf vein and avoid egg-laying, generating a scar that also fossilizes. For the first time, a scar produced by the ovipositor has been identified, indicating the existence of undesirable areas for oviposition. Accordingly, we recognize that Coenagrionidae damselflies (narrow-winged damselflies or pond damselflies) have been avoiding leaf veins for at least 52 million years.

Use of Aedes aegypti Oviposition Surveillance and a Geographic Information System for Planning Anti-Vectorial Measures.

Arboviruses transmitted by Aedes aegypti pose a threat to global public health. Because there are no vaccines or drugs available, the prevention of these diseases in Argentina is based on integrated vector control. In this work, the spatiotemporal dynamics of the vector distribution was analyzed by monitoring oviposition. This information allowed the planning of anti-vector interventions and the evaluation of their effect on the relative abundance of mosquito populations in San Ramón de la Nueva Orán. Observed data were compared with the eggs expected via a statistical model based on meteorological variables. The oviposition substrate preference of mosquito females was also evaluated, and the possible relationship between the relative abundance of the vector and sociodemographic and environmental variables (normalized difference vegetation index and normalized difference water index) was explored. A total of 4,193 eggs of Ae. aegypti were collected, and spatial clusters were detected for all months for which the presence of the mosquito was reported. The observed number of eggs was significantly less than the expected-corrected egg abundance. A significant correlation of oviposition activity was found with three sociodemographic variables, whereas no significant correlation was found with mean or median values of the environmental variables studied. This monitoring strategy made it possible to address the interventions and evaluate them, proposing them as good complementary tools for the control of Ae. aegypti in northern Argentina.

1,2,3-Triazolyl-tetrahydropyrimidine Conjugates as Potential Sterol Carrier Protein-2 Inhibitors: Larvicidal Activity against the Malaria Vector Anopheles arabiensis and In Silico Molecular Docking Study.

Alteration of insect growth regulators by the action of inhibitors is becoming an attractive strategy to combat disease-transmitting insects. In the present study, we investigated the larvicidal effect of 1,2,3-triazolyl-pyrimidinone derivatives against the larvae of the mosquito Anopheles arabiensis, a vector of malaria. All compounds demonstrated insecticidal activity against mosquito larvae in a dose-dependent fashion. A preliminary study of the structure-activity relationship indicated that the electron-withdrawing substituent in the para position of the 4-phenyl-pyrimidinone moiety enhanced the molecules' potency. A docking study of these derivatives revealed favorable binding affinity for the sterol carrier protein-2 receptor, a protein present in the intestine of the mosquito larvae. Being effective insecticides against the malaria-transmitting Anopheles arabiensis, 1,2,3-triazole-based pyrimidinones represent a starting point to develop novel inhibitors of insect growth regulators.

Larvicidal Activities of 2-Aryl-2,3-Dihydroquinazolin -4-ones against Malaria Vector Anopheles arabiensis, In Silico ADMET Prediction and Molecular Target Investigation.

Malaria, affecting all continents, remains one of the life-threatening diseases introduced by parasites that are transmitted to humans through the bites of infected Anopheles mosquitoes. Although insecticides are currently used to reduce malaria transmission, their safety concern for living systems, as well as the environment, is a growing problem. Therefore, the discovery of novel, less toxic, and environmentally safe molecules to effectively combat the control of these vectors is in high demand. In order to identify new potential larvicidal agents, a series of 2-aryl-1,2-dihydroquinazolin-4-one derivatives were synthesized and evaluated for their larvicidal activity against Anopheles arabiensis. The in silico absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties of the compounds were also investigated and most of the derivatives possessed a favorable ADMET profile. Computational modeling studies of the title compounds demonstrated a favorable binding interaction against the acetylcholinesterase enzyme molecular target. Thus, 2-aryl-1,2-dihydroquinazolin-4-ones were identified as a novel class of Anopheles arabiensis insecticides which can be used as lead molecules for the further development of more potent and safer larvicidal agents for treating malaria.

Geometric morphometrics of endophytic oviposition traces of Odonata (Eocene, Argentina).

The insertion of the Odonata ovipositor in the plant tissue generates a scar that surrounds the eggs (trace). In insects, individual egg traces are known to vary in size, but their variation in individual shape is mostly unknown. Twenty-four specimens were obtained from the Laguna del Hunco (Lower Eocene, Chubut) and Río Pichileufú (Middle Eocene, Río Negro), Argentina, which had 1346 oviposition traces (MEF Collection). For the first time, a study of the shape and size of a large number of individual Odonata endophytic egg traces was carried out using traditional (general and mixed linear models) and geometric morphometrics (Fourier elliptical series) to elucidate whether there are changes in size or shape of the individual endophytic egg traces associated with the substrate used at the time of oviposition, if the Lower Eocene traces have varied in relation to those of the Middle Eocene, and if the ichnological classification (Paleoovoidus arcuatus, P. bifurcatus and P. rectus) reflects such variations. We found differences in size (p < 0.05), but not in shape, in relation to the variables studied. This could reflect that the shape of Odonata eggs (inferred from the traces), unlike their size, could have a strong evolutionary constraint already observed since the Eocene.

Redescription of immatures of Dasyhelea flavifrons Guérin-Méneville (Culicomorpha: Ceratopogonidae) and new contribution to the knowledge of its larval habitats.

The fourth instar larva and pupa of Dasyhelea flavifrons Guérin-Méneville are redescribed, illustrated, and photomicrographed using binocular, phase-contrast, and scanning electron microscopy. Comparisons with the American species of the grisea group were made. The immatures were collected by using a siphon bottle in tree-holes and from water collected in dead snail shells in Salta Province, Argentina, transported to the laboratory and there bred to the emergence of the adults. Details on larval habitats are given. These are the first records from Argentina and in gastrotelmata.

Vegetation Cover and Microspatial Distribution of Sand Flies (Diptera: Psychodidae) in an Endemic Locality for Cutaneous Leishmaniasis in Northern Argentina.

The sand fly fauna in Hipólito Yrigoyen, Argentina, a locality where cutaneous leishmaniasis cases occur, was surveyed with zones of higher abundance of sand flies correlated to vegetation cover estimated through normalized difference vegetation index (NDVI). Sand flies were collected with 10 CDC traps during six nights, from December 2009 to January 2010. A map was built of expected sand flies abundance in which levels of NDVI were categorized. In total, 1,392 Phlebotominae (Diptera: Psychodidae) specimens were collected, comprised of the following species: Nyssomyia neivai (Pinto 1926), Migonemyia migonei (França 1920), species of the cortelezzii complex (Brèthes 1923), Evandromyia sallesi (Galvão & Coutinho 1940), and Psathyromyia shannoni (Dyar 1929). Positive correlations were found between the abundance of sand flies and the NDVI (P < 0.05) for buffer areas of <150 m radii from the trap location points, i.e., the sand fly abundance was greater where vegetation cover and density were greater. In this context, plant cover should be taken into account to prioritize surveillance and control areas within the program of sand flies control in northern Argentina.

Larvicidal study of tetrahydropyrimidine scaffolds against Anopheles arabiensis and structural insight by single crystal X-ray studies.

A series of methyl or ethyl 4-(substitutedphenyl/pyridyl)-6-methyl-2-oxo/thioxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate (HPM) analogues 4a-g were synthesized and evaluated for larvicidal activity against Anopheles arabiensis. These newly synthesized compounds were characterized by spectral studies such as FT-IR, NMR (1 H and 13 C), LC-MS, and elemental analysis. The conformational features and supramolecular assembly of molecules 4a, 4b, and 4e were further analyzed from single crystal X-ray study. The larvicidal activity of these tetrahydropyrimidine pharmacophore series was analyzed based on their relative substituents. Among the synthesized HPM analogous from the series, compounds 4d and 4e both having electron withdrawing chlorine group on phenyl ring at the fourth position of the tetrahydropyrimidine pharmacophore exhibited the most promising larvicidal activity.

Evaluation of Multiple Immersion Effects on Eggs From Haemagogus leucocelaenus, Haemagogus janthinomys, and Aedes albopictus (Diptera: Culicidae) Under Experimental Conditions.

Studies on the bioecology of Haemagogus leucocelaenus Dyar and Shannon 1924, Haemagogus janthinomys Dyar 1921, Aedes albopictus Skuse 1895 (Diptera: Culicidae) mosquitos are extremely important from an epidemiologic point of view, as they are known to be vectors of many important pathogens and, therefore, act as the main factor responsible for the maintenance of several zoonoses natural cycles. The present work aimed to elucidate their seasonal egg-hatching rate using the immersion method. Ovitraps were used to collect mosquito eggs from an Atlantic Forest fragment, in the State of Rio de Janeiro, Brazil, from November 2015 to November 2016. After collection, the eggs were immersed 40 times to assess their hatching rate and evaluate the number of immersions resulting in the highest hatchability during the study period. Differences in the proportion of hatched eggs between species and seasons (spring, summer, fall, winter) and in the numbers of immersions in which eggs hatched were assessed using odds ratios. Hg. leucocelaenus was the species with the highest number of eggs hatching in all sampling periods, followed by Ae. albopictus. Most Ae. albopictus eggs hatched on first immersion regardless of season. Both the numbers of eggs and the number of immersion in which the Haemagogus eggs hatched showed high variability within seasons. In spring, the proportion of eggs that hatched on the first compared to further immersions was similar, while in fall and winter a higher percentage (over 94%) of Hg. leucocelaenus eggs hatched on the first immersion; the opposite pattern was observed in the summer. These results differ from previous observations linking increased hatching to warmer months. The number of immersions in which Hg. leucocelaenus eggs hatched varied between seasons, however differences were not statistically significant. These results evidence the need for further studies to elucidate factors that influence hatching patterns.

Distribution of Haemagogus and Sabethes Species in Relation to Forest Cover and Climatic Factors in the Chapada Dos Guimarães National Park, State of Mato Grosso, Brazil.

Members of the genera Haemagogus and Sabethes are the most important biological vectors of the wild yellow fever virus (WYF) in the forested areas of the Americas. The ecologies of Haemagogus janthinomys, Hg. leucocelaenus, Sabethes chloropterus, and Sa. glaucodaemon were studied in a forest of the Chapada dos Guimarães National Park, state of Mato Grosso, Brazil, to evaluate the influence of climatic factors (temperature and relative humidity [RH]) on their abundance. We also examined the association of climate with landscape structure on species distribution patterns throughout the seasons of the year. Multiple stepwise regressions showed that RH was most likely to influence the density of mosquito populations. A multidimensional scaling (MDS) was used to evaluate the effects of forest cover on the composition of mosquito populations at different radii (100-, 250-, and 1,000-m-radius buffer areas). The MDS provided 2 dimensions with values that indicated a higher similarity in the composition of culicid populations between sites 1 and 3, while site 2 was separate from the others in the ordination space. Site 2 had a much higher forest cover ratio at 100-m radius compared with sites 1 and 3. We found a possible relationship between the forest cover and the composition of the mosquito populations only in the 100-m radius. These results enabled us to infer that RH directly favored the activity of mosquito populations and that the forest cover located closest to the sampling site may influence the species composition. Since mosquito abundance was higher in the sites with lower local forest cover, forest fragmentation may be a key factor on the presence of WYF vector.

Synthesis and characterization of a novel series of 1,4-dihydropyridine analogues for larvicidal activity against Anopheles arabiensis.

The new-fangled bis(4-substituted benzyl) 4-(4-substitued phenyl)-2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylate derivatives were synthesized by the union of substituted aryl aldehyde, tert-butyl acetoacetate, ammonium carbonate with 4-substituted benzyl alcohol via Hantzsch ester synthesis in aqueous medium under catalyst-free conditions. The newly synthesized compounds were characterized by spectroscopic techniques such as IR, NMR (1 H and 13 C), ESI mass, elemental analysis, and single-crystal X-ray diffraction. The characterized title compounds were evaluated for the larvicidal activity against Anopheles arabiensis by standard WHO larvicidal assay method using Temephos as standard at 4 µg/ml. The title compounds bis(4-methoxybenzyl) 2,6-dimethyl-4-(4-nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate and bis(4-chlorobenzyl) 2,6-dimethyl-4-(4-nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate exhibited promising larvicidal activity at 65.6% and 72.2%, respectively, when compared with the standard compound at 98.9%.

Snail shells as larval habitat of Limatus durhamii (Diptera: Culicidae) in the Yungas of Argentina.

The shells of dead snails collect water from rainfalls producing aquatic microenvironments called gastrotelmata. These habitats are small and hold simple detritus based on animal communities, being rotifers and culicids the most studied. Although a high diversity of aquatic microhabitats has been reported as larval habitats of mosquitoes in Argentina, the shell of snails has not been investigated yet. We report the shells of three species of native Megalobulimus genus as larval habitats of a neotropical mosquito and suspected vector of bunyaviruses, Limatus durhamii, and describe these microhabitats in the Yungas forest of Argentina.

Membrane effects of dihydropyrimidine analogues with larvicidal activity.

Two recently synthesized dihydropyrimidines (DHPMs) analogues have demonstrated larvicide and repellent activity against Anopheles arabiensis. DHPMs high lipophilicity suggests that these compounds may interact directly with the membrane and modify their biophysical properties. The purpose of the present study was to characterize the interaction of both compounds with artificial membranes. Changes on the properties of DPPC films were studied using Langmuir monolayers. The presence of DHPMs in the subphase modified the interfacial characteristics of DPPC compression isotherms, causing the expansion of the monolayer, inducing the disappearance of DPPC phase transition and increasing the molecular packing of the film. Moreover, both compounds showed ability to penetrate into the lipid monolayers at molecular pressures comparable to those in biological membranes. The effects of both DHPMs on the molecular organization of DPPC liposomes were measured by fluorescence anisotropy. The results indicate that their presence between lipid molecules would induce an increasing intermolecular interaction, diminishing the bilayer fluidity mainly at the polar region. Finally, we performed free diffusion MD simulations and obtained spatially resolved free energy profiles of DHPMs partition into a DPPC bilayer through Potential of Mean Force (PMF) calculations. In agreement with the experimental assays, PMF profiles and MD simulations showed that DHPMs are able to partition into DPPC bilayers, penetrating into the membrane and stablishing hydrogen bonds with the carbonyl moiety. Our results suggest that DHPMs bioactivity could involve their interaction with the lipid molecules that modulate the supramolecular organization of the biological membranes and consequently the membrane proteins functionality.

Biodiversity and Temporal Distribution of Immature Culicidae in the Atlantic Forest, Rio de Janeiro State, Brazil.

To increase the knowledge of biodiversity and identify larval habitats used by immature mosquitoes in the Atlantic Forest, we conducted a study in areas with various stages of preservation within the Guapiaçu Ecological Reserve in Cachoeiras de Macacu, Rio de Janeiro state. The Culicidae fauna were sampled during February, April, June, August, October, and December 2012; February, March, April, May, June, August, October, and December 2013; and January and March 2014. Immature mosquitoes were collected with dippers and suction tubes (mouth aspirators). Over the sampling period, 2697 larvae of 56 species were collected, some of which are recognized vectors of human diseases. The larval mosquito community found in artificial habitats, temporary ground water, and phytotelmata differed between sites, except for the mosquito fauna in bromeliads, which were almost 80% similar. Species segregation was more evident between larval habitats than between sites. Culex usquatus was the dominant species and colonized the highest number of larval habitats. The artificial larval habitats found in REGUA were colonized by a great diversity of species and high abundance as well, thus human artifacts left by the public in the area that collect water may promote an increase in mosquito populations. Among the species collected, some are known or suspected vectors of pathogens to humans and/or veterinary relevance, and their medical relevance is discussed.

Greener synthesis of indolizine analogues using water as a base and solvent: study for larvicidal activity against Anopheles arabiensis.

Greener synthesis of a series of novel indolizine analogues have been achieved by the cyclization of aromatic cycloimmonium ylides with electron-deficient alkynes in the presence of water as the base and solvent at 80 °C. Yield of the title compounds was good and reactions performed were eco-friendly. The structures of these newly synthesized compounds have been confirmed by spectroscopic techniques such as FTIR, NMR, LC-MS, and elemental analysis. Characterized title compounds were evaluated for larvicidal activity against Anopheles arabiensis by standard WHO larvicidal assay using Temefos as standard at 4 µg/mL. Title compounds 2e, 2f, and 2g emerged as promising larvicidal agents.

Vertical oviposition activity of mosquitoes in the Atlantic Forest of Brazil with emphasis on the sylvan vector, Haemagogus leucocelaenus (Diptera: Culicidae).

This study aimed to assess the vertical patterns of oviposition and temporal changes in the distribution of mosquito species in an area of the Atlantic Forest in Rio de Janeiro State, Brazil, and in particular, the behavior and oviposition of potential yellow fever virus vectors. Mosquito samples were collected from the Ecological Reserve Guapiaçu (REGUA, Brazil), which includes a somewhat disturbed forest, with a large diversity of plants and animals. In all, 5,458 specimens (ten species from seven genera) were collected. Haemagogus leucocelaenus was the most frequently captured species, representing 73% of the specimens collected. Species richness and diversity were the highest in the samples collected from the ground-level ovitraps and decreased with height. Species composition also differed significantly among heights. The largest species differences were detected between ovitraps set at the ground level and those set at 7 m and 9 m; Hg. leucocelaenus, Limatus durhamii, and Limatus paraensis contributed most to these differences. Sampling month and climatic variables had significant effects on species richness and diversity. Species diversity and richness decreased with height, suggesting that the conditions for mosquito breeding are more favorable closer to the ground. Species composition also showed vertical differences.