Bamboo (Poales, Poaceae): An Important Maintainer of Immature Mosquitoes (Diptera: Culicidae) in a Biodiversity Hotspot in the City of Rio de Janeiro, Brazil.

Although tropical forests are home to most of the global diversity, they suffer from the most significant knowledge gaps concerning their fauna. Despite its high biodiversity, Brazil is facing an alarming destruction of habitats, with species becoming extinct before they can be discovered or described via science. Therefore, there is an urgent need to expand wildlife inventories, including entomofauna surveys. The present study aimed to analyze the bionomic aspects and the influence of abiotic factors on mosquito fauna whose immature phases develop in two bamboo species, Guadua tagoara and Bambusa vulgaris, in Tijuca National Park, Rio de Janeiro, Brazil. Immatures were collected in 10 artificially drilled bamboo plants, in five stalk internodes per plant, at two sampling points, from March 2022 to March 2023, during 23 collections. A total of 1845 immatures were obtained, 72.14% at sampling point 1 and 27.86% at sampling point 2. Of this, 1162 individuals reached adulthood, belonging to the following species: Culex iridescens, Culex neglectus, Haemagogus leucocelaenus, Orthopodomyia albicosta, Sabethes identicus, Sabethes melanonymphe, Sabethes purpureus, Toxorhynchites bambusicola, Toxorhynchites sp., Trichoprosopon compressum, Trichoprosopon pallidiventer, Wyeomyia arthrostigma, Wyeomyia codiocampa, Wyeomyia lutzi, Wyeomyia oblita, Wyeomyia personata, Wyeomyia serrata, and Wyeomyia sp. The Tijuca National Park is a tourist spot and receives a large number of visitors. Thus, humans can become an accessible food source for mosquitoes in this area, making the species survey critical since important arbovirus vectors have been recorded in Rio de Janeiro.

Ecobiology of Haemagogus leucocelaenus arbovirus vector in the golden lion tamarin translocation area of Rio de Janeiro, Brazil.

Significant pathogens that have resurfaced in humans originate from transmission from animal to human populations. In the Americas, yellow fever cases in humans are usually associated with spillover from non-human primates via mosquitoes. The present study characterized the prevalence of the yellow fever vector Haemagogus leucocelaenus in Rio de Janeiro, Brazil. The Atlantic Forest fragment chosen is an area of translocation of the golden lion tamarin (Leontopithecus rosalia), where 10 ovitraps were installed to collect mosquito eggs in Fazenda Três Irmãos, at Silva Jardim city, from March 2020 to October 2022. A total of 1514 eggs were collected, of which 1153 were viable; 50% belonged to medically important mosquito species and 24% to the yellow fever vector species, Hg. leucocelaenus. The months of December 2020 (n = 252), November 2021 (n = 188), and January 2022 (n = 252) had the highest densities of this vector. Haemagogus leucocelaenus was positively correlated with temperature (r = 0.303) and humidity (r = 0.48), with eggs hatching up to the 15th immersion with higher abundance of females. Implementing mosquito monitoring for arbovirus activity can help protect both the golden lion tamarin and human populations from the threat of arbovirus transmission.

Vertical Distribution of Oviposition and Temporal Segregation of Arbovirus Vector Mosquitoes (Diptera: Culicidae) in a Fragment of the Atlantic Forest, State of Rio de Janeiro, Brazil.

Culicid species, which include potential vectors of yellow fever, are diverse and abundant, with species commonly co-occurring in certain sites. Studying these species can provide important insights into their vector potential and, consequently, epizootic cycles of arboviruses carried about by these vectors. Here, we evaluated the vertical distribution and temporal segregation of mosquito oviposition with emphasis on arbovirus vectors in a fragment of the Atlantic Forest in Casimiro de Abreu, Rio de Janeiro, Brazil. Two sampling points were selected: Fazenda Três Montes and the Reserva Natural de Propriedade Privada Morro Grande. Collections were carried out at two sites using 10 ovitraps installed on the vegetation cover at different heights (0, 2, 4, 6, and 8 m above ground level) and monitored monthly from July 2018 to December 2020. The hypotheses of temporal and vertical stratification were tested through a PERMANOVA, and the relationship of each species with the vertical distribution was evaluated individually through a correlation analysis. We collected a total of 3075 eggs, including four species of medical importance: Haemagogus leucocelaenus (n = 1513), Haemagogus janthinomys (n = 16), Aedes albopictus (n = 1097), and Aedes terrens (n = 449). We found that Hg. leucocelaenus had a positive relationship with height, exhibiting behavior that appears to benefit from higher heights. The abundance of Ae. terrens seemed to follow Hg. leucocelaenus, although we did not find a relationship with height for the former species. On the other hand, Ae. albopictus exhibited a negative relationship with height, becoming absent or outnumbered at higher strata. Our study site has already presented evidence of recent transmission of the wild yellow fever virus, supporting the need to carefully monitor the emergence of febrile diseases among residents in the surrounding areas and the local population.

Occurrence of the Sylvatic Yellow Fever Vector Mosquito Haemagogus leucocelaenus (Diptera: Culicidae) in an Atlantic Forest Fragment of the Touristic State of Rio de Janeiro, Brazil.

The yellow fever virus is estimated to cause 30,000 deaths each year worldwide, with the majority of cases and deaths occurring in Africa. The virus is also endemic to Central and South America, including northern and western Brazil. The sylvatic cycle of the virus is related to wild and rural areas, with nonhuman primates as the primary host and wild mosquitoes, specifically from the genera Haemagogus, as vectors. The diversity of the mosquito community plays a significant role in the increase of pathogen transmission to humans. In the present study, we detected fluctuation in populations of vector mosquitoes using ovitraps for Culicidae egg collection. The study area is a forest fragment of the Atlantic Forest, one of the most threatened biomes in Brazil. This biome has been suffering significant deforestation due to anthropic activity. Worryingly, the proximity of human populations to forest environments increases the risk of spreading disease from forest fragments to urban areas. Our findings showed that the highest egg abundance occurred in December 2019, with a significant difference (p = 0.005) between rainy and dry seasons. Most eggs were collected during the rainy period. Subsequent quantification of specimens from epidemiologically relevant species hatched from field-collected eggs resulted in 1,131 (86%) Haemagogus leucocelaenus (Dyar & Shannon, 1924), 111 (8%) Aedes terrens (Walker, 1856), 47 (4%) Aedes albopictus (Skuse, 1894), and 21 (2%) Haemagogus janthinomys (Dyar, 1921). Finally, we assessed the behavior of different vector species performing oviposition on the same breeding site. The highest correlation coefficient was observed between Ae. albopictus and Ae. terrens (rho = 0.52) concerning other Culicidae species. Therefore, we believe that Culicidae population surveillance is crucial for disease monitoring since the increase in specimens of a number of vector species influences the emergence of yellow fever cases in nonhuman primates and human populations.

Report of Mosquito Vectors of Arboviruses from a Federal Conservation Unit in the Atlantic Forest, Rio de Janeiro State, Brazil.

Arbovirus infections, such as dengue, zika, chikungunya, and yellow fever, are a major public health problem worldwide. As the main vectors, mosquitoes have been classified by the Center for Disease Control and Prevention as one of the deadliest animals alive. In this ecological study, we analyzed the population dynamics of important genera and species of mosquito vectors. Mosquito immatures were collected using ovitraps and at natural breeding sites: bamboos and bromeliads. Adult mosquitoes were captured using CDC traps with CO2, Shannon traps, and manual suction tubes. Collections took place during the rainy and dry seasons from 2019 to 2020 in the Serra dos Órgãos National Park, Rio de Janeiro state, Brazil. The highest number of species was recorded in the ovitraps, followed by CDC and bromeliads. The breeding site with the lowest diversity was bamboo, though it showed the highest level of evenness compared to the other breeding sites. The medically important genera reported were Haemagogus spp., Aedes spp., Culex spp., and Wyeomyia spp. Culicid eggs increased in the rainy season, with a peak in November 2019 and January and February 2020, and lower abundance in the dry season, from September to October 2019. Mosquito eggs had a strong positive correlation (ρ = 0.755) with temperature and a moderate positive correlation (ρ = 0.625) with rainfall. This study shows how environmental variables can influence the ecology of disease-vector mosquitoes, which are critical in the maintenance of arbovirus circulation in a threatened biome within the most densely populated region of Brazil.

Spatial distribution and interactions between mosquitoes (Diptera: Culicidae) and climatic factors in the Amazon, with emphasis on the tribe Mansoniini.

This work aimed to evaluate the spatial distribution of mosquitoes in different seasonal periods and the interaction between climatic factors and the abundance of mosquitoes, especially those belonging to the tribe Mansoniini in the area surrounding the Amazon hydroelectric production region (Jirau-HP) of Rondônia state, Brazil. Mosquito specimens were collected in May, July, October, and December 2018, and April, July, September, and November 2019, over periods of three alternating days during the hours of 6:00 p.m. to 8:00 p.m. Mosquito sampling was performed using automatic CDC and Shannon light traps. Canonical correspondence analysis (CCA), combined with Monte Carlo permutations, was used to evaluate the correlation between climatic variables and species distribution. In addition, non-metric multidimensional scaling (NMDS) was used to verify the similarity among the sampled communities from the different collections. After analyzing the total mosquito fauna at all sampling points, 46,564 specimens were identified, with Mansonia dyari showing the highest relative abundance in 2018 (35.9%). In contrast, Mansonia titillans had the highest relative abundance in 2019 (25.34%), followed by Mansonia iguassuensis (24.26%). The CCA showed that maximum temperature significantly influenced the distribution of mosquito populations in the study area (p = 0.0406). The NMDS showed that sampling carried out in the rainy and dry seasons formed two distinct groups. There was a significant correlation between species richness and cumulative precipitation 15 days before the sampling period (R2 = 58.39%; p = 0.0272). Thus, both temperature and precipitation affected mosquito population dynamics. The effect of rainfall on mosquito communities may be due to variations in habitat availability for immature forms.

Little noticed, but very important: The role of breeding sites formed by bamboos in maintaining the diversity of mosquitoes (Diptera: Culicidae) in the Atlantic Forest biome.

This study investigated the composition of mosquito species in different kinds of breeding sites in a tropical forest remnant of the Atlantic Forest and identified species of public health concern therein. Collections of immature forms of mosquitoes were carried out monthly at the Poço das Antas Biological Reserve in southeastern Brazil, between June 2014 and June 2015. Samples were collected from four types of breeding sites: bamboos, bromeliads, puddles, and a lake. A total of 1,182 specimens of mosquitoes belonging to 28 species and 13 genera were collected. Three species, Ad. squamipennis, An. neglectus, and Wy. arthrostigma represented 64.8% of the captured specimens. Only three species were found in more than one type of breeding site: Ps. ferox, An. triannulatus, and Tx. trichopygus. Two species of public health concern were found breeding in bamboo (Ae. aegypti and Ae. albopictus) and one in the lake (An. darlingi). Bamboo had the highest species richness, Shannon diversity, abundance of individuals and number of dominant species of all breeding sites. Similar Simpson diversity was obtained for bamboo and bromeliads, with higher values than those obtained for puddles and the lake. The significance of the four breeding sites, especially bamboos, is discussed in the context of controlling populations of sylvatic species of mosquitoes in Atlantic Forest areas.

Interaction of Haemagogus leucocelaenus (Diptera: Culicidae) and Other Mosquito Vectors in a Forested Area, Rio de Janeiro, Brazil.

The yellow fever (YF) virus has been detected throughout Brazil, with the occurrence of human cases, cyclic epizootics, and its isolation from Haemagogus janthinomys and Hg. leucocelaenus. We assessed the seasonal occurrence, egg abundancy, and oviposition interaction of mosquito vector species captured at a Private Natural Heritage Reserve in the Atlantic Forest biome. A total of 2943 eggs and 1538 mosquito larvae were collected from which 1231 belonged to entomologically important species. Ovitraps were used to collect immature mosquitoes from September 2019 to January 2021. The Mann-Whitney test was used to assess the differences in the abundance of eggs between rainy and dry seasons. Kruskal-Wallis and Dunn's post hoc tests were used to evaluate the significance of the differences in the number of individuals from vector species. The highest percentage of mosquito vector eggs were collected during the rainy season, from December to February. Most eggs recovered from ovitraps belonged to the species Hg. leucocelaenus, representing 85% of all mosquito eggs identified. The other species had lower abundances and percentages: Aedes terrens (7%), Haemagogus janthinomys (5%) and Aedes albopictus (3%). The species that shared breeding sites with a higher frequency were Hg. leucocelaenus and Hg. janthinomys, with a statistically positive correlation (ρ = 0.74). This finding suggests that maybe the presence of Hg. leucocelaenus eggs acted as an attractant for Hg. janthinomys or vice versa. An understanding of mosquito oviposition behavior is necessary for the development of surveillance and control approaches directed against specific pathogen vectors of medical and veterinary importance.

Influence of seasonality on wing morphological variability in populations of Mansonia amazonensis (Theobald) (Diptera: Culicidae).

Mansonia amazonensis (Theobald, 1901) is one of 15 species of the subgenus Mansonia Blanchard, 1901. It is essentially a Neotropical species, recorded in Bolivia, Peru and Brazil. In the highly seasonal Amazon region, Mansonia species find ideal environmental conditions for reproduction, development and establishment. Considering that climate significantly influences the size and behavior of mosquitoes, and insects in general, we analyzed the influence of seasonality on wing morphological variability in populations of Ma. amazonensis. Captures were carried out near the banks of the Madeira River in Rondnia State, Brazil during the dry, rainy and transition periods between seasons during 2018 and 2019. Morphometric characters of 181 wings were analyzed using morphological methods. The results show that wing size of Ma. amazonensis increases following a relatively gradual trend, from smaller wings during the dry period to larger ones in the rainy season. This study provides the first evidence, detected using geometric morphometry, of seasonally associated phenotypic variability in the wing conformation of Ma. amazonensis.

Diversity of Mosquitoes (Diptera: Culicidae) in the Bom Retiro Private Natural Heritage Reserve, Rio de Janeiro State, Brazil.

This study registers the diversity of Culicidae in the Bom Retiro Private Natural Heritage Reserve (RPPNBR), Rio de Janeiro state, Brazil, based on the collection of the immature stages in natural and artificial larval habitats. Larvae and pupae were collected monthly at two sites of the RPPNBR from May 2014 to July 2015 using dippers and aquatic pipettes. The diversity of the mosquito community was described using the Shannon-Wiener Diversity Index (H'), as well as diversity, richness, and dominance of species found in different larval habitats (lake, bamboos, bromeliads, and artificial vessels). The Mann-Whitney test was used to calculate differences between the two natural and artificial habitats. Overall, 15,659 specimens belonging to 25 species, ten genera, and two subfamilies were collected. The most abundant species collected at sites that were reforested recently were Culex pleuristriatus Theobald, 1903, Limatus durhamii (Theobald, 1901), Aedes albopictus (Skuse, 1895), Culex neglectus (Lutz, 1904), and Culex retrosus (Lane & Whitman, 1951). In a forest preserved site, the most abundant species were Cx. neglectus, Culex iridescens (Lutz, 1905), Sabethes identicus (Dyar & Knab, 1907), Wyeomyia arthrostigma (Lutz, 1905), and Li. durhamii. With respect to larval habitats, 0.1% of the specimens were collected along the edge of a lake, 5.5% in bamboos, 35.9% in bromeliads, and 58.4% in artificial containers. Only 5.5% of the specimens were collected in the forest preserved site, with the remaining samples from the site with altered vegetation. A greater species richness and diversity were found in forest-altered sites compared to the forest preserved site. Several species were collected in the water accumulated in the nylon lids of plastic water tanks. Such vessels can promote an increase in mosquito population density in the environment surrounding the study area.

Sexual Proportion and Egg Hatching of Vector Mosquitos in an Atlantic Forest Fragment in Rio de Janeiro, Brazil.

Some Aedinii mosquitoes are of high importance in the transmission of the sylvatic YFV. Usually, their eggs are very resistant and depend on the rain for their hatching. The present study evaluated the effect of multiple mosquito-egg immersions and the sex ratio of male and female specimens from Atlantic Forest remnants in the state of Rio de Janeiro, Brazil. Three sampling sites were selected in the municipality of Casimiro de Abreu, where 50 ovitraps were randomly installed to collect eggs from the ground level up to different heights, from August 2018 to December 2020. The mosquito sex ratios were compared between seasons and forest sites, using the generalized linear mixed model (GLMM), which included sampling months and trees as random effects. A total of 33,091 mosquito eggs were collected, of which 6152 eggs were already hatched (18%) and 26,939 were unhatched; of these, approximately 76% subsequently hatched. We found that 25% of the eggs corresponded to four species: Aedes albopictus (n = 1277), Ae. terrens (n = 793), Haemagogus janthinomys (n = 89), and Hg. leucocelaenus (n = 3033). The sex ratio (male:female) was variable concerning the sampling sites and the season. For most species, GLMM estimates found no difference in the variation of the average sex ratio as a function of these predictors, and there was no evidence of temporal autocorrelation in the mosquito data. The number of immersions necessary for hatching the eggs differed between mosquito species, and eggs collected in the dry season hatched both in the first immersions and the subsequent events. Co-occurrence of Aedes terrens and Hg. leucocelaenus was the most frequently observed pairwise species combination. Considering recurrent arbovirus outbreaks in Brazil and their burden on the human population, our study helps to shed light on how these vectors behave in nature; therefore, they can be used in surveillance programs.

Oviposition activity of Haemagogus leucocelaenus (Diptera: Culicidae) during the rainy and dry seasons, in areas with yellow fever virus circulation in the Atlantic Forest, Rio de Janeiro, Brazil.

The present study aims to analyze the effectiveness of ovitraps in the capture of Hg leucocelaenus eggs and evaluate the influence of the dry and rainy seasons on their abundance and hatching rates. The eggs were collected in the Atlantic Forest of State of Rio de Janeiro, Brazil, an area in which the yellow fever virus is known to circulate. We distributed 15 ovitraps in three sampling points, with five ovitraps per point. We distributed 15 ovitraps in three sampling points on trees within a forested area, which were sequentially numbered, monitored, and replaced every two weeks from October 2016 to April 2018. There was a high dominance of Hg. leucocelaenus eggs (98.4%) and a variation in egg hatching rates between the wet and dry seasons. These rates were 1.5 times higher in the rainy season than in the dry season. The rainy season also showed a greater abundance of eggs and higher values of ovitrap positivity and egg density indexes in the installed ovitraps. The abundances of Hg. leucocelaenus eggs were positively correlated with mean monthly temperature and air humidity but not significantly correlated with accumulated precipitation. These results, as well as their implications for the possible use of ovitraps to monitor vector mosquitoes of yellow fever in the study region, are discussed.

Dispersion pattern of Mansonia in the surroundings of the Amazon Jirau Hydroelectric Power Plant.

Mansonia spp. are voracious hematophagous mosquitoes whose mature stages usually breed in freshwater bodies containing aquatic vegetation. The reduction in water flow leads to a proliferation in aquatic plants, increasing their populations. Besides, some species are potential vectors of pathogens such as arboviruses and microfilariae. We evaluated the degree of active dispersion of females of Mansonia spp. in the surrounding area of the Jirau hydroelectric power plant in the Amazon, Rondônia, Brazil, using mark-release-recapture techniques. The flight behavior of the recaptured specimens was summarized with a set of average and maximum distances traveled. We show that the dispersal movement of Mansonia spp. is predominantly performed by random, low, and short flights, with a tendency to remain near the breeding sites in certain vegetation fragments. However, the maximum distances traveled were 2000 m from the release point for Mansonia amazonensis during 2018 and Mansonia humeralis during 2019.

Spawning behavior of Aedini (Diptera: Culicidae) in a remnant of Atlantic Forest in the state of Rio de Janeiro.

BACKGROUND: Mosquito assemblages are organized along an ecological gradient, including small habitats where interspecific competition predominates and large permanent habitats where predation predominates. This study aimed to analyze the oviposition behavior of mosquitoes regarding the preference for traps installed at two different heights with regard to ground level and the tendency to share spawning sites in an Atlantic Forest fragment in Nova Iguaçu, State of Rio de Janeiro, Brazil. METHODS: The eggs were collected from April 2018 to March 2019. Twelve ovitraps were used, randomly distributed in trees at ground level and at a height of 3 m in a forest environment. RESULTS: They were sequentially numbered, monitored, and replaced every 2 weeks. Among the 5818 eggs collected, 3941 hatched, 3756 reached the pupa stage, and 2370 reached the adult stage. The most abundant species were Aedes albopictus (63%) and Haemagogus leucocelaenus (35%), followed by Aedes terrens (2%) and Haemagogus janthinomys (1%). Analyses showed a significant difference of (P = 0.02) between the number of mosquito species collected in the palettes at ground level and the number collected at the height of 3 m. Cluster analysis of species abundance showed that the eggs collected in the palettes at ground level were more abundant than those collected in the palettes at the height of 3 m. We detected co-occurrence of species in the oviposition palettes; according to the null model, such species distribution was not random. CONCLUSIONS: The exploitation of oviposition sites by mosquito species can represent an event forced by population density facilitated by the ecological valence of individuals of one species. Understanding the aggregate distribution of larvae at the oviposition site allows us to conduct more in-depth studies of the oviposition behavior of female mosquitoes.

Bionomy of Mosquitoes in Bamboo Internodes in an Atlantic Forest Remnant of the State of Rio De Janeiro, Brazil.

Living bamboo stalks are one of the most specialized habitats for mosquito oviposition and immature development. Most of the mosquito species that breed in these habitats are sylvatic, and some are of importance for public health as possible vectors of pathogens. Perforated internodes are a very specialized environment due to the difficulty of access. Furthermore, due to their relatively simple fauna, they represent a valuable model for ecological studies that may be applicable to more complex environments. This study aims to assess the mosquito bionomics of species raised in bamboo internodes. Therefore, the diversity of mosquito species and the influence of abiotic variables (pH and temperature) on the distribution of mosquitoes that breed in this habitat were analyzed. The study area is a fragment of Atlantic Forest within the Association of da Armada (ATA) in Nova Iguaçu, state of Rio de Janeiro, Brazil. Immature mosquitoes were sampled with suction tubes (mouth aspirators) between August 2017 and July 2018. A total of 3,170 larvae were collected in 5 bamboo plants, each with 8 stalks perforated. Of these, 688 larvae reached the adult stage, representing 10 genera and 19 species. The most common species were Culex neglectus (43%), Trichoprosopon digitatum (22%), Culex iridescens (8%), Sabethes identicus (7%), and Orthopodomyia albicosta (7%). The richness of the immatures collected in the ATA was 19 species, with a diversity of 1.10 and Shannon evenness of 0.57. A diverse composition of Culicidae in bamboo stalks was found, although dominance was low.

Oviposition behavior of wild yellow fever vector mosquitoes (Diptera: Culicidae) in an Atlantic Forest fragment, Rio de Janeiro state, Brazil.

Although there are many studies on the control of mosquito vectors of the yellow fever virus (YFV) in tropical forests, there are still few ecological studies regarding abiotic factors effect on these mosquitoes. Here we characterize these effects on oviposition behavior, abundance, and diversity of mosquito vectors of YFV. The study was conducted in Córrego da Luz Municipal Park, in Casimiro de Abreu, Rio de Janeiro state, Brazil, from July 2018 to December 2019. Ovitraps were placed at ground level and 3 m high. The data were tested for normality using the Shapiro-Wilk test, followed by an independent sample analysis, the Mann-Whitney test. The Shannon Diversity Index was used to evaluate the abundance of mosquitos' eggs collected at both ground level and 3 m high. We highlight the presence of Haemagogus janthinomys and Hg. leucocelaenus, primary YFV vectors in forest areas. The abundance of Hg. leucocelaenus (63%), Hg. janthinomys (75%), and Aedes terrens (58%) was higher at the height of 3 m, while Ae. albopictus (52%) was higher at ground level. Aedes albopictus was positively correlated with temperature. Culicidae monitoring is essential for assessing the YFV transmission cycle in Atlantic forest fragments.

Ultrastructure of the egg of Coquillettidia venezuelensis (Theobald) (Diptera: Culicidae).

This study examined the ultrastructural characteristics of the egg of Coquillettidia venezuelensis (Theobald) (Diptera: Culicidae) with the aid of scanning electron microscopy. The eggs are elliptical and blackish, measuring on average 525.3 ± 12.8 µm in length and 94.9 ± 5.14 µm in width, with a length to width ratio of 5.54 ± 0.18. The anterior region of the egg is tubuliform, with a well-developed micropylar collar measuring approximately 2.54 ± 0.33 µm in thickness, the inner side of which is uniformly and deeply excavated, with a depression around the outer margin of the micropylar disc, which has a diameter of 29.4 ± 1.7 µm, an area of 234.4 µm2 ± 401.3 µm2 and a slightly elevated central region measuring 1.7 ± 0.5 µm in diameter that bears the central micropyle.

Ultrastructure and morphometry of the egg of Coquillettidia albifera (Prado) with illustrations of male genitalia (Diptera: Culicidae).

The ultrastructure and morphometry of the egg of Coquillettidia albifera (Prado) (Diptera: Culicidae) studied under scanning electronic microscopy is presented for the first time. The identification of the species was confirmed by examination of the genitalia of males collected with females. The male genitalia are redescribed and digital photographs are provided. The eggs are elliptical with the anterior region elongate and tubuliform, length 661 ± 2.59 µm and width 183 ± 2.59 µm (ratio length/width 3.61 ± 0.08). The dorsal surface is covered by a hexagonal, pentagonal and rectangular chorionic network with randomly distributed tubercles. The micropylar disc has a diameter of 18.91 ± 0.65 µm, surrounded by a continuous protuberant collar with a thickness of 4.4 ± 0.3 µm. No structure or vestige of cement related to egg attachment was observed.

Description of the egg of Hulecoeteomyia koreica (Edwards) (Diptera: Culicidae) using scanning electron microscopy.

The egg of Hulecoeteomyia koreica (Edwards) [=Aedes koreicus (Edwards)], an invasive mosquito in Europe, is described from material obtained in Italy. Eggs are cigar-shaped and tapered posteriorly. They are 667.5 ±47.27 µm in length, and the width posteriorly, in the middle and near the micropyle is, respectively, 113.18 ±10.07, 182.5 ±26.92 and 142.25 ±13.85µm. The outer chorion has groups of tubercles, 3 or 4 in an irregular row in the anterior region and 4-6 irregularly dispersed in cavities surrounded by a covering similar to a membrane. The micropyle is surrounded by an irregular collar, and groups of tubercles are less defined in this region. This pattern is different from the egg described for any other mosquito species.

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.