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DeGennaro, M.

Publications and source records attributed to DeGennaro, M..

13 recordsLinked to original sources

Odor and bacterial signatures for humans that attract different mosquito species

Humans are not equally attractive to mosquitoes, leaving some more vulnerable to mosquito-borne illnesses than others. Body odor differences likely allow mosquitoes to discriminate between humans. Using a uniport olfactometer, we measured the attraction of Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus mosquitoes for each of our 119 participants. Ae. aegypti, but not other species tested, were more attracted to male than female participants. Each of our three species ranked our participants differently, favoring a distinct subset of our cohort. For each species, mosquito attraction rates were used to define high and low attraction human odors and bacterial taxa. For example, Ae. aegypti and Cx. quinquefasciatus attraction was associated with the absence of odors like cyclic alcohols and monoterpenes, while Ae. albopictus attraction was associated with the presence of ketones. Each mosquito species exhibited distinct responses to individual humans, emphasizing both unique and shared cues for targeting their hosts.

neuroscience↗

Unraveling forensic timelines using molecular markers in Phormia regina maggots

In forensic entomology, estimating the time of death is critical and traditionally relies on changes in observable traits of carrion feeding insect larvae. Traits such as size, weight, and morphology can be used to predict the insect specimen age and help estimate the time since death. The blowfly Phormia regina is a key forensic insect, yet age estimation for older maggots in this species is particularly challenging due to the limited morphological changes in the late-stage larvae. To enhance precision, we employed transcriptomic profiling on blowfly maggots, aiming to identify genes as markers for time of death estimation. Our study characterized maggot development, reinforcing that weight and behavior cannot precisely determine age between 100 and 130 hours. We built a chromosomal scale annotated genome, establishing a reliable database for uncovering transcriptomic signatures during larval development. Applying differential gene expression analyses, weighted gene co-expression network analysis, and the generalized linear model, we identified nine candidate genes (Y5078, Y5076, agt2, ech1, dhb4, asm, gabd, acohc, Ivd) that delineate the age of otherwise indeterminate maggots. This research introduces a molecular approach to address a longstanding problem in forensic entomology and promises to increase precision in determining the time of death at a crime scene. AUTHOR SUMMARYIn forensic entomology, estimating the time of death often depends on insect morphological changes. The blowfly Phormia regina is a key species, but aging older maggots in the third instar larval phase is difficult due to minimal changes in observable traits. To improve accuracy, we used transcriptomic profiling to identify genetic markers for time of death estimation. Our study showed that weight and behavior are not reliable indicators of age in maggots 100 to 130 hours old. We developed an annotated genome to identify transcriptomic signatures during larval development. Through various analyses, we identified nine genes (Y5078, Y5076, agt2, ech1, dhb4, asm, gabd, acohc, Ivd) that can determine the age of late third instar maggots. This molecular approach aims to enhance precision in forensic entomology.

developmental biology↗

Landscape and meteorological variables associated with Aedes aegypti and Aedes albopictus mosquito infestation in two southeastern USA coastal cities

Aedes transmitted arboviral human cases are increasing worldwide and spreading to new areas of the United States of America (USA). These diseases continue to re-emerge likely due to changes in vector ecology, urbanization, human migration, and larger range of climatic suitability. Recent shifts in landscape and weather variables are predicted to impact the habitat patterns of urban mosquitoes such as Aedes aegypti and Aedes albopictus. Miami (FL) is in the tropical zone and an established hotspot for arboviruses, while Charleston (SC) is in the humid subtropical zone and newly vulnerable. Although these coastal cities have distinct climates, both have hot summers. To understand mosquito infestation in both cities and potentiate our surveillance effort, we performed egg collections in the warmest season. We applied remote sensing with land-use cover and weather variation to identify mosquito infestation patterns. Our study found predominant occurrence of Ae. aegypti and, to a lesser extent, Ae. albopictus in both cities. We detected statistically significant positive and negative associations between entomological indicators and most weather variables in combined data from both cities. For all entomological indices, weekly wind speed and relative humidity were significantly positively associated, while precipitation and maximum temperature were significantly negatively associated. Aedes egg abundance was significantly positively associated with open land in Charleston but was negatively associated with vegetation cover in combined data. There is a clear need for further observational studies to determine the impact of climate change on Ae. aegypti and Ae. albopictus infestation in the Southeastern region of the USA.

ecology↗

Global genomics of Aedes aegypti unveils widespread and novel persistent infectious viruses capable of triggering a small RNA response

The mosquito Aedes aegypti is a prominent vector for arboviruses, but the breadth of mosquito viruses that infects this specie is not fully understood. In the broadest global survey to date of over 200 Ae. aegypti small RNA samples, we detected viral small interfering RNAs (siRNAs) and Piwi interacting RNAs (piRNAs) arising from mosquito viruses. We confirmed that most academic laboratory colonies of Ae. aegypti lack persisting viruses, yet two commercial strains were infected by a novel tombus-like virus. Ae. aegypti from North to South American locations were also teeming with multiple insect viruses, with Anphevirus and a bunyavirus displaying geographical boundaries from the viral small RNA patterns. Asian Ae. aegypti small RNA patterns indicate infections by similar mosquito viruses from the Americas and reveal the first wild example of dengue virus infection generating viral small RNAs. African Ae. aegypti also contained various viral small RNAs including novel viruses only found in these African substrains. Intriguingly, viral long RNA patterns can differ from small RNA patterns, indicative of viral transcripts evading the mosquitoes RNA interference (RNAi) machinery. To determine whether the viruses we discovered via small RNA sequencing were replicating and transmissible, we infected C6/36 and Aag2 cells with Ae. aegypti homogenates. Through blind passaging, we generated cell lines stably infected by these mosquito viruses which then generated abundant viral siRNAs and piRNAs that resemble the native mosquito viral small RNA patterns. This mosquito small RNA genomics approach augments surveillance approaches for emerging infectious diseases.

genomics↗

Fertility decline in Aedes mosquitoes is associated with reduced maternal transcript deposition and does not depend on female age

Female mosquitoes undergo multiple rounds of reproduction known as gonotrophic cycles. A gonotrophic cycle spans the period from bloodmeal intake to egg laying. Nutrients from vertebrate host blood are necessary for completing egg development. During oogenesis, a female pre-packages mRNA into her oocytes, and these maternal transcripts drive the first two hours of embryonic development prior to zygotic genome activation. In this study, we profiled transcriptional changes in 1-2 hours old Aedes aegypti embryos across two gonotrophic cycles. We found that homeotic genes which are regulators of embryogenesis are downregulated in embryos from the second gonotrophic cycle. Interestingly, embryos produced by Ae. aegypti females progressively reduced their ability to hatch as the number of gonotrophic cycles increased. We show that this fertility decline is due to increased reproductive output and not the mosquitoes age. Moreover, we found a similar decline in fertility and fecundity across three gonotrophic cycles in Ae. albopictus. Our results are useful for predicting mosquito population dynamics to inform vector control efforts.

developmental biology↗

Cross-modal sensory compensation increases mosquito attraction to humans

Sensory compensation is a process that allows individuals with a loss of one sense, for instance hearing or vision, to adapt to changes in their sensory abilities. Where this phenomenon has been observed, there is enhanced perception by another sense to compensate for deficiency of the lost sense. Such compensation is important for humans and non-human animals that use multisensory integration for effective navigation and the execution of vital tasks. Among these, female mosquitoes are sensory specialists that rely heavily on integrating multiple human-emitted cues in their quest for a suitable host to obtain a blood meal. Here, we identify a previously undescribed mechanism of sensory compensation in female Aedes aegypti mosquitoes. Mutant mosquitoes lacking the odorant receptor co-receptor Orco show specific enhancement in heat-seeking behavior. This s compensation does not require the antenna, which was previously assumed to be the primary mosquito thermosensitive organ. Instead, we found that the tips of the forelegs are required to detect heat, and that the heightened sensitivity in heat detection is mediated by increased neuronal activity in foreleg sensory neurons, which are distant from the head appendage neurons that express Orco. By comparative gene expression analysis in wildtype and Orco mutant legs, we identify Ir140, a foreleg-enriched member of the Ionotropic Receptor (IR) superfamily of sensory receptors, as strongly upregulated in Orco mutant legs. Emphasizing the important role of IRs in thermosensation, we find that mutant mosquitoes lacking the IR co-receptor, Ir25a, lose all responses to heat, and Ir140 mutants show strong deficits in responding to human skin temperatures. We generated an Ir140, Orco double mutant and show that these animals lose the remarkable sensory compensation seen in Orco mutants. This strongly suggests that upregulation of Ir140 in the foreleg is the mechanism of sensory compensation in Orco mutants. Odorant receptor expression is sparse in legs, suggesting an indirect, long-range mechanism of sensory compensation. Our findings reveal a novel compensatory mechanism in which loss of one sensory modality in female Aedes aegypti mosquitoes results in greater sensitivity in another to maintain the overall effectiveness of their host-seeking behavior, further enhancing their status as the most dangerous predator of humans.

neuroscience↗

Odor-evoked transcriptomics of Aedes aegypti mosquitoes

Modulation of odorant receptors mRNA induced by prolonged odor exposure is highly correlated with ligand-receptor interactions in Drosophila as well as mammals of the Muridae family. If this response feature is conserved in other organisms, this presents a potentially potent initial screening tool when searching for novel receptor-ligand interactions in species with predominantly orphan olfactory receptors. We demonstrate that mRNA modulation in response to 1-octen-3-ol odor exposure occurs in a time- and concentration-dependent manner in Aedes aegypti mosquitoes. To investigate gene expression patterns at a global level, we generated an odor-evoked transcriptome associated with 1-octen-3-ol odor exposure. Transcriptomic data revealed that ORs and OBPs were transcriptionally responsive whereas other chemosensory gene families showed little to no differential expression. Alongside chemosensory gene expression changes, transcriptomic analysis found that prolonged exposure to 1-octen-3-ol modulated xenobiotic response genes, primarily members of the cytochrome P450, insect cuticle proteins, and glucuronosyltransferases families. Together, these findings suggest that mRNA transcriptional modulation caused by prolonged odor exposure is pervasive across taxa and accompanied by the activation of xenobiotic responses. Furthermore, odor-evoked transcriptomics create a potential screening tool for filtering and identification of chemosensory and xenobiotic targets of interest.

neuroscience↗

Fertility decline in female mosquitoes is regulated by the orco olfactory co-receptor

Female Aedes aegypti mosquitoes undergo multiple rounds of reproduction, known as gonotrophic cycles. These cycles span the period from blood meal intake to oviposition. Understanding how reproductive success is maintained across gonotrophic cycles allows for the identification of molecular targets to reduce mosquito population growth. Odorant receptor co-receptor (orco) encodes a conserved insect-specific transmembrane ion channel that complexes with tuning odorant receptors (ORs) to form a functional olfactory receptor. orco expression has been identified in the male and female mosquito germline, but its role is unclear. We report an orco-dependent, maternal effect reduction in fertility after the first gonotrophic cycle. This decline was rescued by repairing the orco mutant locus. Eggs deposited by orco mutant females are fertilized but the embryos reveal developmental defects, reduced hatching, and changes in ion channel signaling gene transcription. We present an unexpected role for an olfactory receptor pathway in mosquito reproduction. HIGHLIGHTSO_LILoss of the orco olfactory co-receptor promotes female mosquito fertility decline. C_LIO_LIAfter their first reproductive cycle completes, Ae. aegypti orco mutant females produce embryos with disrupted development and reduced hatching. C_LIO_LICRISPR/Cas9-mediated repair of the orco mutation rescues the fertility defect. C_LIO_LIGene expression profiling of embryos from orco mutant females supports a role for ion channel signaling in mosquito development. C_LI

genetics↗

Food recognition in a blood-feeding insect: characterization of the pharyngeal taste organ

BackgroundObligate blood-feeding insects obtain the nutrients and water necessary to ensure survival from the vertebrate blood. The internal taste sensilla, situated in the pharynx, evaluate the suitability of the ingested food. Here, through multiple approaches, we characterized the pharyngeal organ (PO) of the hematophagous kissing bug Rhodnius prolixus to determine its role in food assessment. The PO, located antero-dorsally in the pharynx, comprises 8 taste sensilla that become bathed with the incoming blood. ResultsWe showed that these taste sensilla house gustatory receptor neurons projecting their axons through the labral nerves to reach the subesophageal zone in the brain. We found that these neurons are electrically activated by relevant appetitive and aversive gustatory stimuli such as NaCl, ATP and caffeine. Using RNA-Seq, we examined the expression of sensory-related gene families in the PO. We identified gustatory receptors, ionotropic receptors, transient receptor potential channels, pickpocket channels, opsins, takeouts, neuropeptide precursors, neuropeptide receptors and biogenic amine receptors. RNA interference assays demonstrated that the pickpocket channel Rproppk014276 is necessary for salt detection during feeding. ConclusionWe provide evidence of the role of the pharyngeal organ in food evaluation. This work shows the first comprehensive characterization of a pharyngeal taste organ in a hematophagous insect.

neuroscience↗

Carboxylic acids that drive mosquito attraction to humans activate ionotropic receptors

The mosquito, Aedes aegypti, is highly anthropophilic and continues to transmit debilitating arboviruses within human populations. Female mosquitoes are attracted to sources of blood by responding to odor plumes that are emitted by their preferred hosts. Acidic volatile compounds, including carboxylic acids, represent particularly salient odors driving this attraction. Importantly, carboxylic acids are major constituents of human sweat and volatiles generated by skin microbes. As such, they are likely to impact human host preference, which is a dominant factor in disease transmission cycles. A more complete understanding of mosquito host attraction will necessitate the elucidation of molecular mechanisms of volatile odor detection that function in peripheral sensory neurons. Recent studies have shown that members of the variant ionotropic glutamate receptor gene family are necessary for physiological and behavioral responses to acidic volatiles in Aedes. In this study, we have identified a subfamily of variant ionotropic receptors that share sequence homology across several important vector species and are likely to be activated by carboxylic acids. Moreover, we demonstrate that selected members of this subfamily are activated by short chain carboxylic acids in a heterologous cell expression system. Our results are consistent with the hypothesis that members of this receptor class underlie acidic volatile sensitivity in vector mosquitoes and provide a frame of reference for future development of novel mosquito attractant and repellent technologies.

molecular biology↗

Symbiosis-driven development in an early branching metazoan

Microbes can initiate developmental gene regulatory cascades in animals. The molecular mechanisms underlying microbe-induced animal development and the evolutionary steps to integrate microbial signals into regulatory programs remain poorly understood. In the upside-down jellyfish Cassiopea xamachana, a dinoflagellate endosymbiont initiates the life stage transition from the sessile polyp to the sexual medusa. We found that metabolic products derived from symbiont carotenoids may be important to initiate C. xamachana development, in addition to expression of conserved genes involved in medusa development of non-symbiotic jellyfish. We also revealed the transcription factor COUP is expressed during metamorphosis, potentially as a co-regulator of nuclear receptor RXR. These data suggest relatively few steps may be necessary to integrate symbiont signals into gene regulatory networks and cements the role of the symbiont as a key trigger for life history transition in C. xamachana.

developmental biology↗

Increased temperatures reduce the vectorial capacity of Aedes mosquitoes for Zika virus

Rapid and significant range expansion of both ZIKV and its Aedes host species has resulted in ZIKV being declared a global health threat. Mean temperatures are projected to increase globally, likely resulting in alterations of the transmission potential of mosquito-borne pathogens. The relationship between temperature and ZIKV transmission has not been well characterised for Aedes aegypti and Aedes albopictus.\n\nTo understand the effect of diurnal temperature range on the vectorial capacity of Aedes aegypti and Aedes albopictus for ZIKV, factors contributing to transmission potential were measured at different temperature regimens. Their longevity and blood feeding rates were assessed, and vector competence was determined following feeding on blood meals with 8.3 log10 PFU/ml ZIKV.\n\nHigher temperature resulted in decreased longevity of Ae. aegypti [Log-rank (Mantel-Cox) Test, Chi-square, df 35.66, 5 P (<0.0001)] and a significant decrease in blood feeding rates across groups [Z score (-5.8478) P (0.0444)]. Temperature had a population and species-specific impact on ZIKV infection rates. Overall, Ae. albopictus reared at the lowest temperature regimen demonstrated the highest vectorial capacity (1.63) and the highest transmission efficiency (57%). Temperature increases decreased vectorial capacity across groups, yet the largest decreases were measured for Ae. aegypti.\n\nThe results of this study suggest that future climate change could significantly impact vector competence, blood feeding behavior and longevity, and therefore decrease the overall vectorial capacity of Aedes mosquitoes. It is also clear that this impact is likely to be both species and population-specific.

microbiology↗

Geosmin attracts Aedes aegypti mosquitoes to oviposition sites

Geosmin is one of the most recognizable and common microbial smells on the planet. Some insects, like mosquitoes, require microbial-rich environments for their progeny, whereas for other insects such microbes may prove dangerous. In the vinegar fly Drosophila melanogaster, geosmin is decoded in a remarkably precise fashion and induces aversion, presumably signaling the presence of harmful microbes. We have here investigated the effect of geosmin on the behavior of the yellow fever mosquito Aedes aegypti. In contrast to flies, geosmin is not aversive in mosquitoes but stimulates egg-laying site selection. Female mosquitoes could associate geosmin with microbes, including cyanobacteria consumed by larvae, who also find geosmin - as well as geosmin producing cyanobacteria - attractive. Using in vivo multiphoton imaging from mosquitoes with pan-neural expression of the calcium reporter GCaMP6s, we show that Ae. aegypti code geosmin in a similar fashion to flies, i.e. with extreme sensitivity and with a high degree of selectivity. We further demonstrate that geosmin can be used as bait under field conditions, and finally we show that geosmin, which is both expensive and difficult to obtain, can be substituted by beetroot peel extract, providing a cheap and viable mean of mosquito control and surveillance in developing countries.

neuroscience↗