The surveillance of pre-mRNA splicing is an early step in C. elegans RNAi of endogenous genes.

RNAi pathways detect and silence foreign nucleic acids such as viruses as well as endogenous genes in many species. The phylogenetic profile across eukaryotes of proteins that mediate key steps in RNAi is correlated with the profiles of multiple mRNA splicing proteins and with intron number, suggesting that RNAi may surveil mRNA splicing to detect the divergent or absent introns of viruses. Here we examine the role of mRNA splicing in Caenorhabditis elegans RNAi. We found that viable null mutations in U1 and U2 small nuclear ribonucleic protein (snRNP)-specific splicing factor genes cause defects in RNAi. The U1A ortholog rnp-2 is required for normal ERGO-1 Argonaute class 26G siRNA biogenesis, trans-splicing of the eri-6/7 transcript, and targeting of poorly conserved gene transcripts by WAGO Argonaute class 22G siRNAs. We found that gene transcripts engaged by the siRNA-generating machinery are poorly conserved, possess few introns, and often have introns that are divergent from introns with strong consensus splicing sites found in highly conserved genes. We present biochemical evidence that RNAi targeted transcripts are tightly bound to spliceosomes. These findings suggest multiple layers of regulation by the spliceosome at early steps of small RNA-mediated gene silencing.

Assessment of Mayaro virus vector competence of the mosquito Aedes aegypti (Linnaeus, 1762) populations in Argentine using dose-response assays.

Mayaro virus (MAYV; Alphavirus: Togaviridae) is an emerging pathogen in Latin America, causing fever and polyarthritis. Sporadic outbreaks of MAYV have occurred in the region, with reported human cases being imported to Europe and North America. Although primarily a risk for those residing in the Amazon basin's tropical forests, recent reports highlight that urbanization would increase the risk of MAYV transmission in Latin America. Urban emergence depends on human susceptibility and the ability of mosquitos like Aedes aegypti  (Linnaeus, 1762) (Diptera: Culicidae) to transmit MAYV. Despite the absence of active MAYV transmission in Argentine, the risk of introduction is substantial due to human movement and the presence of Ae. aegypti in the region. This study aimed to evaluate the susceptibility of different Argentine Ae. aegypti populations to MAYV genotype L (MAYV-L) using dose-response assays and determine barriers to virus infection, dissemination and transmission. Immature mosquito stages were collected in Buenos Aires, Córdoba and Rosario cities. Female Ae. aegypti (F2) were orally infected by feeding on five concentrations of MAYV-L, ranging from 1.0 to 6.0 log10 PFU/mL. Abdomens, legs and saliva were analysed using viral plaque assays. Results revealed that MAYV-L between infection and dissemination were associated with viral doses rather than the population origin. Infection rates varied between 3% and 65%, with a 50% infectious dose >5.5 log10 PFU/mL. Dissemination occurred at 39%, with a 50% dissemination dose of ~6.0 log10 PFU/mL. Dissemination among infected mosquitoes ranged from 60% to 86%, and transmission from disseminated mosquitoes ranged from 11% to 20%. Argentine Ae. aegypti populations exhibited a need for higher viral doses of MAYV-L than those typically found in humans to become infected. In addition, only a small proportion of infected mosquitoes were capable of transmitting the virus. Understanding MAYV transmission in urban areas is crucial for public health interventions.

Caenorhabditis elegans ADAR editing and the ERI-6/7/MOV10 RNAi pathway silence endogenous viral elements and LTR retrotransposons.

Endogenous retroviruses and long terminal repeat (LTR) retrotransposons are mobile genetic elements that are closely related to retroviruses. Desilenced endogenous retroviruses are associated with human autoimmune disorders and neurodegenerative diseases. Caenorhabditis elegans and related Caenorhabditis spp. contain LTR retrotransposons and, as described here, numerous integrated viral genes including viral envelope genes that are part of LTR retrotransposons. We found that both LTR retrotransposons and endogenous viral elements are silenced by ADARs [adenosine deaminases acting on double-stranded RNA (dsRNA)] together with the endogenous RNA interference (RNAi) factor ERI-6/7, a homolog of MOV10 helicase, a retrotransposon and retrovirus restriction factor in human. siRNAs corresponding to integrated viral genes and LTR retrotransposons, but not to DNA transposons, are dependent on the ADARs and ERI-6/7. siRNAs corresponding to palindromic repeats are independent of the ADARs and ERI-6/7, and are in fact increased in adar- and eri-6/7-defective mutants because of an antiviral RNAi response to dsRNA. Silencing of LTR retrotransposons is dependent on downstream RNAi factors and P granule components but is independent of the viral sensor DRH-1/RIG-I and the nuclear Argonaute NRDE-3. The activation of retrotransposons in the ADAR- and ERI-6/7/MOV10-defective mutant is associated with the induction of the unfolded protein response (UPR), a common response to viral infection. The overlap between genes induced upon viral infection and infection with intracellular pathogens and genes coexpressed with retrotransposons suggests that there is a common response to different types of foreign elements that includes a response to proteotoxicity presumably caused by the burden of replicating pathogens and expressed retrotransposons.

Behavior of Aedes albifasciatus (Diptera: Culicidae) larvae from eggs with different dormancy times and its relationship with parasitism by Strelkovimermis spiculatus (Nematoda: Mermithidae).

Aedine mosquitoes go through unfavorable periods as dormant eggs. However, extended dormancy times lead to a depletion of egg reserves, which might be partially compensated by changes in larval-feeding behavior. Changes in larval behavior may also be induced by parasitism by mermithids probably as a strategy to reduce the impact of the parasite on the host. The most abundant floodwater mosquito in southern South America is Aedes albifasciatus (Diptera: Culicidae), a species naturally parasitized by Strelkovimermis spiculatus (Nematoda: Mermithidae). This study aimed to evaluate the behavior of fourth-instar larvae of Ae. albifasciatus from eggs with 2, 4, and 6 months of dormancy, parasitized or not by S. spiculatus. To achieve this, eight categories of behavior were defined, and then, each individual was observed for 5 min, and its behaviors and their duration were recorded. The behaviors with the highest percentage of observation time were still (50.1%) and wriggle swimming (33.6%), while those with the lowest percentage of observation time were those involving active foraging (less than 8%). A great variability in the behaviors was observed among individuals within the same treatment. The type and duration of each behavior did not vary in relation to egg dormancy time or parasitism, but parasitism affected the level of larval activity. Parasitized larvae performed, on average, fewer behaviors than non-parasitized larvae. This result supports the hypothesis that S. spiculatus parasitism affects the behavior of Ae. albifasciatus larvae by reducing their level of activity.

Differential inhibition of egg hatching in Aedes aegypti populations from localities with different winter conditions.

In Argentina, the mosquito Aedes aegypti (L.) (Diptera: Culicidae) is distributed from subtropical to temperate climates. Here, we hypothesized that the expansion of Ae. aegypti into colder regions is favoured by high-phenotypic plasticity and an adaptive inhibition of egg hatching at low temperatures. Thus, we investigated the hatching response of eggs of three populations: one from a subtropical region (Resistencia) and two from temperate regions (Buenos Aires City and San Bernardo) of Argentina. Eggs collected in the field were raised in three experimental colonies. F1 eggs were acclimated for 7 days prior to immersion at 7.6 or 22°C (control eggs). Five immersion temperatures were tested: 7.6, 10.3, 11.8, 14.1 and 16°C (range of mean winter temperatures of the three localities). A second immersion at 22°C was performed 2 weeks later to assess the inhibition to hatch under favourable conditions. After the first immersion, we compared the proportions of hatched eggs and dead larvae among treatment levels, whereas after the second immersion we compared the hatching response among the three populations. The factors that most influenced the egg hatching response were the geographical origin of the populations and the immersion temperature, but not the acclimation temperature. The proportions of hatching and larval mortality at low temperatures were higher for Resistencia than for Buenos Aires and San Bernardo, whereas the hatching response at ambient temperature was lower for San Bernardo than for Buenos Aires and Resistencia. The results support the hypothesis that populations from colder regions show an adaptive inhibition of egg hatching.

Delayed mortality effects of cold fronts during the winter season on Aedes aegypti in a temperate region.

The expansion of the invasive mosquito Aedes aegypti L. (Diptera: Culicidae) towards temperate regions in the Americas is causing concern because of its public health implications. As for other insects, the distribution limits of Ae. aegypti have been suggested to be related to minimum temperatures and to be controlled mainly by cold tolerance. The aim of this study was to assess the daily mortality of immature stages of Ae. aegypti under natural winter conditions in Buenos Aires, Argentina, in relation to preceding thermal conditions. The experiment was performed outdoors, and one cohort of larvae was started each week for 16 weeks, and reared up to the emergence of the adults. Three times a week, larvae, pupae and emerged adults were counted, and these data were used to calculate the daily mortality of larvae, pupae and adults and to analyze their relationship with thermal conditions. The results showed that mortality was generally low, with a few peaks of high mortality after cold front events. The mortality of pupae and larvae showed a higher correlation with the cooling degree hours of previous days than with the minimum, maximum or mean temperatures. Pupae and adults showed to be more vulnerable to low temperatures than larvae. A delay in mortality was observed in relation to the low temperature events, with a proportion of individuals dying in a later stage after the end of the cold front. These results suggest that thermal conditions during cold fronts in Buenos Aires are close to the tolerance limit of the local Ae. aegypti population. The wide range of responses of different individuals suggests that low winter temperatures may constitute a selective force, leading the population to a higher tolerance to low temperatures, which might favor the further expansion of this species towards colder regions.

Multiple small RNA pathways regulate the silencing of repeated and foreign genes in C. elegans.

Gene segments from other organisms, such as viruses, are detected as foreign and targeted for silencing by RNAi pathways. A deep-sequencing map of the small RNA response to repeated transgenes introduced to Caenorhabditis elegans revealed that specific segments are targeted by siRNAs. Silencing of the foreign gene segments depends on an antiviral response that involves changes in active and silent chromatin modifications and altered levels of antisense siRNAs. Distinct Argonaute proteins target foreign genes for silencing or protection against silencing. We used a repeated transgene in a genome-wide screen to identify gene disruptions that enhance silencing of foreign genetic elements and identified 69 genes. These genes cluster in four groups based on overlapping sets of coexpressed genes, including a group of germline-expressed genes that are likely coregulated by the E2F transcription factor. Many of the gene inactivations enhance exogenous RNAi. About half of the 69 genes have roles in endogenous RNAi pathways that regulate diverse processes, including silencing of duplicated genes and transposons and chromosome segregation. Of these newly identified genes, several are required for siRNA biogenesis or stability in the oocyte-specific ERGO-1 pathway, including eri-12, encoding an interactor of the RNAi-defective protein RDE-10, and ntl-9/CNOT9, one of several CCR4/NOT complex genes that we identified. The conserved ARF-like small GTPase ARL-8 is required specifically for primary siRNA biogenesis or stability in the sperm-specific ALG-3/4 endogenous RNAi pathway.

A model for the development of Aedes (Stegomyia) aegypti as a function of the available food.

We discuss the preimaginal development of the mosquito Aedes aegypti from the point of view of the statistics of developmental times and the final body-size of the pupae and adults. We begin the discussion studying existing models in relation to published data for the mosquito. The data suggest a developmental process that is described by exponentially distributed random times. The existing data show as well that the idea of cohorts emerging synchronously is verified only in optimal situations created at the laboratory but it is not verified in field experiments. We propose a model in which immature individuals progress in successive stages, all of them with exponentially distributed times, according to two different rates (one food-dependent and the other food-independent). This phenomenological model, coupled with a general model for growing, can explain the existing observations and new results produced in this work. The emerging picture is that the development of the larvae proceeds through a sequence of steps. Some of the steps depend on the available food. While food is in abundance, all steps can be thought as having equal duration, but when food is scarce, those steps that depend on food take considerably longer times. For insufficient levels of food, increase in larval mortality sets in. As a consequence of the smaller rates, the average pupation time increases and the cohort disperses in time. Dispersion, as measured by standard deviation, becomes a quadratic function of the average time indicating that cohort dispersion responds to the same causes than delays in pupation and adult emergence. During the whole developmental process the larva grows monotonically, initially at an exponential rate but later at decreasing rates, approaching a final body-size. Growth is stopped by maturation when it is already slow. As a consequence of this process, there is a slight bias favoring small individuals: Small individuals are born before larger individuals, although the tendency is very weak.

Cold Season Mortality Under Natural Conditions and Subsequent Hatching Response of Aedes (Stegomyia) aegypti (Diptera: Culicidae) Eggs in a Subtropical City of Argentina.

In temperate and subtropical regions, populations of Aedes (Stegomyia) aegypti (L.) survive unfavorable winter conditions in the egg stage. Knowing their survival rates can be of great interest for the health authorities in charge of control activities. In this study, we analyzed the mortality of Ae. aegypti eggs exposed to the cold season as well as their hatching patterns under laboratory conditions in the city of Resistencia, Chaco, Argentina. The mortality rate was 48.6%. No statistically significant differences were observed in the mortality of eggs exposed at different sites. Hatching response differed significantly among the successive postexposure immersions, with the highest proportion of hatched eggs during the first immersion. These results show that the mortality rate of Ae. aegypti eggs exposed to the cold season in a subtropical city of Argentina was higher than those from temperate climate region. The additional mortality of eggs in our study might be related to fungal development (an unexpected event), which was not observed in research in temperate climate. The hatching pattern observed in this study ensures a rapid increase of the population at the beginning of the favorable breeding season, but it also maintains a batch with delayed hatching eggs, posing a risk for the community.

PIWI associated siRNAs and piRNAs specifically require the Caenorhabditis elegans HEN1 ortholog henn-1.

Small RNAs--including piRNAs, miRNAs, and endogenous siRNAs--bind Argonaute proteins to form RNA silencing complexes that target coding genes, transposons, and aberrant RNAs. To assess the requirements for endogenous siRNA formation and activity in Caenorhabditis elegans, we developed a GFP-based sensor for the endogenous siRNA 22G siR-1, one of a set of abundant siRNAs processed from a precursor RNA mapping to the X chromosome, the X-cluster. Silencing of the sensor is also dependent on the partially complementary, unlinked 26G siR-O7 siRNA. We show that 26G siR-O7 acts in trans to initiate 22G siRNA formation from the X-cluster. The presence of several mispairs between 26G siR-O7 and the X-cluster mRNA, as well as mutagenesis of the siRNA sensor, indicates that siRNA target recognition is permissive to a degree of mispairing. From a candidate reverse genetic screen, we identified several factors required for 22G siR-1 activity, including the chromatin factors mes-4 and gfl-1, the Argonaute ergo-1, and the 3' methyltransferase henn-1. Quantitative RT-PCR of small RNAs in a henn-1 mutant and deep sequencing of methylated small RNAs indicate that siRNAs and piRNAs that associate with PIWI clade Argonautes are methylated by HENN-1, while siRNAs and miRNAs that associate with non-PIWI clade Argonautes are not. Thus, PIWI-class Argonaute proteins are specifically adapted to associate with methylated small RNAs in C. elegans.

First Record and Larval Habitat Description of Culex (Melanoconion) pilosus from Buenos Aires Province, Argentina.

Larvae of Culex (Melanoconion) pilosus were collected during February-April 2014 in temporary pools in "Bosques de Ezeiza," a large forested park, near Buenos Aires city, Argentina. This is the first record in Buenos Aires Province, extending the distribution of this species 380 km to the south. Regarding habitat use, Cx. (Mel.) pilosus is a generalist, although a slight association of larval abundances with pools of lower pH and higher vegetation cover was observed. The comparison of larval instars of Cx. (Mel.) pilosus with those of other genera suggests a life-history strategy similar to that of floodwater mosquitoes.

Trans-splicing in C. elegans generates the negative RNAi regulator ERI-6/7.

Mutations that enhance the response to double-stranded RNA (dsRNA) have revealed components of the RNA interference (RNAi) pathway or related small RNA pathways. To explore these small RNA pathways, we screened for Caenorhabditis elegans mutants displaying an enhanced response to exogenous dsRNAs. Here we describe the isolation of mutations in two adjacent, divergently transcribed open reading frames (eri-6 and eri-7) that fail to complement. eri-6 and eri-7 produce separate pre-messenger RNAs (pre-mRNAs) that are trans-spliced to form a functional mRNA, eri-6/7. Trans-splicing of eri-6/7 is mediated by a direct repeat that flanks the eri-6 gene. Adenosine to inosine editing within untranslated regions of eri-6 and eri-7 pre-mRNAs reveals a double-stranded pre-mRNA intermediate, forming in the nucleus before splicing occurs. The ERI-6/7 protein is a superfamily I helicase that both negatively regulates the exogenous RNAi pathway and functions in an endogenous RNAi pathway.

The ERI-6/7 helicase acts at the first stage of an siRNA amplification pathway that targets recent gene duplications.

Endogenous small interfering RNAs (siRNAs) are a class of naturally occuring regulatory RNAs found in fungi, plants, and animals. Some endogenous siRNAs are required to silence transposons or function in chromosome segregation; however, the specific roles of most endogenous siRNAs are unclear. The helicase gene eri-6/7 was identified in the nematode Caenorhabditis elegans by the enhanced response to exogenous double-stranded RNAs (dsRNAs) of the null mutant. eri-6/7 encodes a helicase homologous to small RNA factors Armitage in Drosophila, SDE3 in Arabidopsis, and Mov10 in humans. Here we show that eri-6/7 mutations cause the loss of 26-nucleotide (nt) endogenous siRNAs derived from genes and pseudogenes in oocytes and embryos, as well as deficiencies in somatic 22-nucleotide secondary siRNAs corresponding to the same loci. About 80 genes are eri-6/7 targets that generate the embryonic endogenous siRNAs that silence the corresponding mRNAs. These 80 genes share extensive nucleotide sequence homology and are poorly conserved, suggesting a role for these endogenous siRNAs in silencing of and thereby directing the fate of recently acquired, duplicated genes. Unlike most endogenous siRNAs in C. elegans, eri-6/7-dependent siRNAs require Dicer. We identify that the eri-6/7-dependent siRNAs have a passenger strand that is ∼19 nt and is inset by ∼3-4 nts from both ends of the 26 nt guide siRNA, suggesting non-canonical Dicer processing. Mutations in the Argonaute ERGO-1, which associates with eri-6/7-dependent 26 nt siRNAs, cause passenger strand stabilization, indicating that ERGO-1 is required to separate the siRNA duplex, presumably through endonucleolytic cleavage of the passenger strand. Thus, like several other siRNA-associated Argonautes with a conserved RNaseH motif, ERGO-1 appears to be required for siRNA maturation.

mut-16 and other mutator class genes modulate 22G and 26G siRNA pathways in Caenorhabditis elegans.

Argonaute-associated siRNAs and Piwi-associated piRNAs have overlapping roles in silencing mobile genetic elements in animals. In Caenorhabditis elegans, mutator (mut) class genes mediate siRNA-guided repression of transposons as well as exogenous RNAi, but their roles in endogenous RNA silencing pathways are not well-understood. To characterize the endogenous small RNAs dependent on mut class genes, small RNA populations from a null allele of mut-16 as well as a regulatory mut-16(mg461) allele that disables only somatic RNAi were subjected to deep sequencing. Additionally, each of the mut class genes was tested for a requirement in 26G siRNA pathways. The results indicate that mut-16 is an essential factor in multiple endogenous germline and somatic siRNA pathways involving several distinct Argonautes and RNA-dependent RNA polymerases. The results also reveal essential roles for mut-2 and mut-7 in the ERGO-1 class 26G siRNA pathway and less critical roles for mut-8, mut-14, and mut-15. We show that transposons are hypersusceptible to mut-16-dependent silencing and identify a requirement for the siRNA machinery in piRNA biogenesis from Tc1 transposons. We also show that the soma-specific mut-16(mg461) mutant allele is present in multiple C. elegans laboratory strains.

Oviposition site selection and subsequent offspring performance of Aedes aegypti in short- and long-term detritus accumulation conditions.

The "oviposition preference-offspring performance" hypothesis proposes that females lay their eggs in habitats that maximize the fitness of their offspring. The aim of this study was to assess the oviposition site selection by Aedes aegypti females and the success of their larvae in habitats with different detritus accumulation times, under conditions representative of the natural spatial variability of detritus quality and quantity in the Metropolitan Area of Buenos Aires, Argentina. Two experiments were performed, one assessing oviposition site selection and the other analyzing developmental success. In both experiments, two levels of detritus accumulation time were compared, one with short-time detritus accumulation (2 weeks), and the other with long-time detritus accumulation (8 weeks). Naturally fallen detritus was used in both experiments, collected in ten sites across the Metropolitan Area of Buenos Aires. In the oviposition experiment, two contiguous ovitraps corresponding to each accumulation time were placed at each of the ten sites and the number of eggs received for each accumulation time was compared. In the development experiment, always 19 larvae were raised in containers of both accumulation times and overall performance was compared using an integrated index that considers survival, development time and female size. A large variability in the amount of detritus collected at the different sites was observed. The number of eggs was significantly higher in the long-time than in the short-time detritus accumulation containers, and approximately proportional to the amount of detritus in each ovitrap. The performance was not affected by the detritus accumulation time, but a better performance was detected in containers that received a higher amount of organic detritus, regardless of the accumulation time. Leaves were on average the most abundant type of detritus, with an average of 53 % of the total detritus collected. The amount of leaves added 2 weeks before hatching showed a positive effect on larval performance. Our results do not support the "oviposition preference-offspring performance" hypothesis, since Ae. aegypti females laid eggs in containers where larvae did not show a better performance. Moreover, at larval densities related to the number of eggs actually laid in each of the accumulation times, it is expected that the performance would be even worse in the most selected containers, due to the density-dependent effects of crowding. Since the results obtained reflect the natural heterogeneity of the environmental conditions in the region studied, they might be a fairly good indicator of what occurs in natural larval habitats.

Bacteria Are a Major Determinant of Orsay Virus Transmission and Infection in Caenorhabditis elegans.

The microbiota is a key determinant of the physiology and immunity of animal hosts. The factors governing the transmissibility of viruses between susceptible hosts are incompletely understood. Bacteria serve as food for Caenorhabditis elegans and represent an integral part of the natural environment of C. elegans. We determined the effects of bacteria isolated with C. elegans from its natural environment on the transmission of Orsay virus in C. elegans using quantitative virus transmission and host susceptibility assays. We observed that Ochrobactrum species promoted Orsay virus transmission, whereas Pseudomonas lurida MYb11 attenuated virus transmission relative to the standard laboratory bacterial food Escherichia coli OP50. We found that pathogenic Pseudomonas aeruginosa strains PA01 and PA14 further attenuated virus transmission. We determined that the amount of Orsay virus required to infect 50% of a C. elegans population on P. lurida MYb11 compared with Ochrobactrum vermis MYb71 was dramatically increased, over three orders of magnitude. Host susceptibility was attenuated even further in presence of P. aeruginosa PA14. Genetic analysis of the determinants of P. aeruginosa required for attenuation of C. elegans susceptibility to Orsay virus infection revealed a role for regulators of quorum sensing. Our data suggest that distinct constituents of the C. elegans microbiota and potential pathogens can have widely divergent effects on Orsay virus transmission, such that associated bacteria can effectively determine host susceptibility versus resistance to viral infection. Our study provides quantitative evidence for a critical role for tripartite host-virus-bacteria interactions in determining the transmissibility of viruses among susceptible hosts.

Bacteria are a major determinant of Orsay virus transmission and infection in Caenorhabditis elegans.

The microbiota is a key determinant of the physiology and immunity of animal hosts. The factors governing the transmissibility of viruses between susceptible hosts are incompletely understood. Bacteria serve as food for Caenorhabditis elegans and represent an integral part of the natural environment of C. elegans. We determined the effects of bacteria isolated with C. elegans from its natural environment on the transmission of Orsay virus in C. elegans using quantitative virus transmission and host susceptibility assays. We observed that Ochrobactrum species promoted Orsay virus transmission, whereas Pseudomonas lurida MYb11 attenuated virus transmission relative to the standard laboratory bacterial food Escherichia coli OP50. We found that pathogenic Pseudomonas aeruginosa strains PA01 and PA14 further attenuated virus transmission. We determined that the amount of Orsay virus required to infect 50% of a C. elegans population on P. lurida MYb11 compared with Ochrobactrum vermis MYb71 was dramatically increased, over three orders of magnitude. Host susceptibility was attenuated even further in the presence of P. aeruginosa PA14. Genetic analysis of the determinants of P. aeruginosa required for attenuation of C. elegans susceptibility to Orsay virus infection revealed a role for regulators of quorum sensing. Our data suggest that distinct constituents of the C. elegans microbiota and potential pathogens can have widely divergent effects on Orsay virus transmission, such that associated bacteria can effectively determine host susceptibility versus resistance to viral infection. Our study provides quantitative evidence for a critical role for tripartite host-virus-bacteria interactions in determining the transmissibility of viruses among susceptible hosts.

Temporal dynamics of the egg bank of Aedes aegypti (Diptera: Culicidae) in the winter-spring transition in a temperate region.

In temperate regions, the populations of Aedes aegypti (L.) (Diptera: Culicidae) remain in the egg stage during the winter. In Buenos Aires, Argentina, a temperate region in the southern limit of Ae. aegypti distribution, the start of the next reproductive season and the rate of increase of the adult population depend on the egg bank that remains after the winter. This study aimed to analyze the mortality, field-hatching, and survival of eggs exposed to field conditions representative of those that occur in nature. In addition, the post-exposure hatching response of the eggs was assessed. Four egg batches were exposed to natural conditions starting in mid-winter and were recovered progressively after 3, 6, 9, and 12 weeks. One egg batch (initial control) was not exposed in the field and remained under laboratory conditions. After the exposure period, the recovered intact, collapsed, and hatched eggs were counted. Intact eggs were immersed three times in the laboratory to study their hatching response. Progressive increases in the proportion of lost (presumably by predation), dead, and hatched eggs in successive egg batches were recorded. Field-hatching was recorded from late winter onwards. The first hatchings occurred in conditions probably not favorable to complete development into reproductive adults. A progressive decrease in live eggs was observed, with 51% of the recovered eggs alive after 12 weeks of exposure. In the laboratory, the hatching response in the first immersion was low for the initial control and for the eggs exposed for 3 weeks, and increased for successive cohorts. The results confirm that the survival of Ae. aegypti eggs in the winter-spring transition ensures persistence throughout the next favorable season in Buenos Aires City. The observed inhibition to hatch of the first batches might relate to a photoperiod-induced diapause, as observed in previous studies.

Genetic variants that modify neuroendocrine gene expression and foraging behavior of C. elegans.

The molecular mechanisms underlying diversity in animal behavior are not well understood. A major experimental challenge is determining the contribution of genetic variants that affect neuronal gene expression to differences in behavioral traits. In Caenorhabditis elegans, the neuroendocrine transforming growth factor-ß ligand, DAF-7, regulates diverse behavioral responses to bacterial food and pathogens. The dynamic neuron-specific expression of daf-7 is modulated by environmental and endogenous bacteria-derived cues. Here, we investigated natural variation in the expression of daf-7 from the ASJ pair of chemosensory neurons. We identified common genetic variants in gap-2, encoding a Ras guanosine triphosphatase (GTPase)-activating protein homologous to mammalian synaptic Ras GTPase-activating protein, which modify daf-7 expression cell nonautonomously and promote exploratory foraging behavior in a partially DAF-7-dependent manner. Our data connect natural variation in neuron-specific gene expression to differences in behavior and suggest that genetic variation in neuroendocrine signaling pathways mediating host-microbe interactions may give rise to diversity in animal behavior.

Immature mosquitoes associated with urban parklands: implications for water and mosquito management.

The aim of the present study was to compare 2 urban habitat types: pools artificially filled with water from damaged or leaking water pipes (AF) and pools naturally filled by rainwater (NF), with regard to their favorability as breeding sites for mosquitoes. Two study areas were analyzed, 1 for 5 months and the other for 9 months, covering the whole period when AF pools contained water. The AF pools held water during the entire study, and showed lower fluctuations in surface area than NF pools. The AF pools showed higher levels of total mosquitoes and of stagnant-water mosquitoes. The floodwater mosquitoes were numerically (but not significantly) more abundant in NF pools. Nine mosquito species were identified. Habitat type, temperature, and season were significant in explaining the variability in species composition according to the canonical correspondence analysis. The most abundant species were Ochlerotatus albifasciatus (= Aedes albifasciatus, predominantly in NF pools), Culex dolosus, and Cx. pipiens (mainly in AF pools). The latter 2 species differed in their temporal dynamics, with Cx. dolosus associated with lower temperatures and Cx. pipiens with higher temperatures. Overall, the results indicate that although both habitat types harbored immature mosquitoes, the AF pools were more favorable than co-occurring rain pools. Easy-to-implement management actions such as the design of adequate drainage systems and the fast repair of broken pipes will be helpful to reduce the risk of human illness associated with mosquitoes in urban green areas.