When More is Less: Mosquito Population Suppression Using Sterile, Incompatible and Genetically Modified Male Mosquitoes.

The current review of the Sterile Insect Technique (SIT) is motivated by new technologies and the recent renaissance of male release field trials, which is driving an evolution in mosquito control and regulation. Practitioners that are releasing male mosquitoes would do well to learn from past successes and failures, including political and public engagement complications. With examples that include nuanced integrations of the different technologies, e.g., combinations of Wolbachia and irradiation, it is critical that scientists understand and communicate accurately about the technologies, including their evolving management by different regulatory agencies in the USA. Some male release approaches are considered 'pesticides' and regulated by federal and state agencies, while other male release approaches are unregulated. It is important to consider how the new technologies fit with the more 'traditional' chemical applications of adulticides and larvicides. The economics of male release programs are substantially different from traditional control costs, which can be a challenge to their adoption by abatement districts. However, there is substantial need to overcome these complications and challenges, because the problem with invasive mosquitoes grows ever worse with factors that include insecticide resistance, globalization and climate change.

Yes, Irradiated Sterile Male Mosquitoes Can Be Sexually Competitive!

Adequate sexual competitiveness of sterile males is a prerequisite for genetic control methods, including the sterile insect technique. During the past decade several semi-field and open-field trials demonstrated that irradiated male mosquitoes can be competitive.

Enhancement of mosquito trapping efficiency by using pulse width modulated light emitting diodes.

In this study, a light-driving bug zapper is presented for well controlling the diseases brought by insects, such as mosquitoes. In order to have the device efficient to trap the insect pests in off-grid areas, pulse width modulated light emitting diodes (PWM-LED) combined with a solar power module are proposed and implemented. With specific PWM electric signals to drive the LED, it is found that no matter what the ability of catching insects or the consumed power efficiency can be enhanced thus. It is demonstrated that 40% of the UV LED consumed power and 25.9% of the total load power consumption can be saved, and the trapped mosquitoes are about 250% increased when the PWM method is applied in the bug zapper experiments.

Light-Driven Processes Control Both Rhodopsin Maturation and Recycling in Mosquito Photoreceptors.

Many invertebrates carry out a daily cycle of shedding and rebuilding of the photoreceptor's photosensitive rhabdomeric membranes. The mosquito Aedes aegypti shows a robust response, losing nearly all Aaop1 rhodopsin from the rhabdomeric membranes during the shedding process at dawn. Here, we made use of Aaop1 antibodies capable of distinguishing newly synthesized, glycosylated rhodopsin from mature nonglycosylated rhodopsin to characterize the fate of Aaop1 during the shedding and rebuilding processes. The rhabdomeric rhodopsin is moved into large cytoplasmic vesicles at dawn and is subsequently degraded during the standard 12 h daytime period. The endocytosed rhodopsin is trafficked back to the photosensitive membranes if animals are shifted back to dark conditions during the morning hours. During the daytime period, small vesicles containing newly synthesized and glycosylated Aaop1 rhodopsin accumulate within the cytoplasm. At dusk, these vesicles are lost as the newly synthesized Aaop1 is converted to the nonglycosylated form and deposited in the rhabdomeres. We demonstrate that light acts though a novel signaling pathway to block rhodopsin maturation, thus inhibiting the deglycosylation and rhabdomeric targeting of newly synthesized Aaop1 rhodopsin. Therefore, light controls two cellular processes responsible for the daily renewal of rhodopsin: rhodopsin endocytosis at dawn and inhibition of rhodopsin maturation until dusk. SIGNIFICANCE STATEMENT: Organisms use multiple strategies to maximize visual capabilities in different light conditions. Many invertebrates show a daily cycle of shedding the photoreceptor's rhabdomeric membranes at dawn and rebuilding these during the following night. We show here that the Aedes aegypti mosquito possesses two distinct light-driven cellular signaling processes for modulating rhodopsin content during this cycle. One of these, endocytosis of rhabdomeric rhodopsin, has been described previously. The second, a light-activated cellular pathway acting to inhibit the anterograde movement of newly synthesized rhodopsin, is revealed here for the first time. The discovery of this cellular signaling pathway controlling a G-protein-coupled receptor is of broad interest due to the prominent role of this receptor family across all areas of neuroscience.

Comparison of different light sources for trapping Culicoides biting midges, mosquitoes and other dipterans.

The response of Culicoides biting midges, mosquitoes and other dipterans to different wavelengths was evaluated in a farm meadow in northern Spain. A total of 9449 specimens of 23 species of Culicoides, 5495 other ceratopogonids (non-biting midges), 602 culicids and 12428 other mixed dipterans were captured. Centers for Disease Control and Prevention (CDC) suction light traps fitted with five light emitting diodes (LEDs) (white, green, red, blue, ultraviolet) were run for 15 consecutive nights. Significantly more Culicoides were collected in those traps fitted with green, blue or ultraviolet (UV) lights than in red and white-baited LED traps for the most abundant species captured: C. punctatus (37.5%), C. cataneii (26.5%) and C. obsoletus/C. scoticus (20.4%). Similar results were obtained for non-Culicoides ceratopogonids, mosquitoes and other mixed dipterans. Wavelengths in green (570nm) resulted effective for targeting some Culicoides species, culicids and other midges. In a second trial, the effectiveness of 4-W white and UV tubes was compared to traps fitted with UV LED and a standard incandescent light bulb. More specimens of all taxa were collected with fluorescent black light (UV) traps than with the other light sources, except culicids, which were recovered in high numbers from fluorescent white light traps.

Synthesis and photophysical properties of the photoactivatable cationic porphyrin 5-(4-N-dodecylpyridyl)-10,15,20-tri(4-N-methylpyridyl)-21H,23H-porphyrin tetraiodide for anti-malaria PDT.

This article describes a new synthetic method for obtaining three water soluble porphyrins. The more sophisticated porphyrin [5-(4-N-dodecylpyridyl)-10,15,20-tri(4-N-methylpyridyl)-21H,23H-porphyrin tetraiodide], also named C12 porphyrin, was obtained through a three step methodology. The improvements, compared to syntheses described in the literature, mostly concern the purification procedures. The photophysical properties of the three porphyrins are described and the C12 porphyrin presents a very good (1)O2 yield compared to its chemical intermediates. This porphyrin seems to be a very promising candidate for PDT applications.

Boosting the sterile insect technique to control mosquitoes.

Mosquitoes are vectors of major diseases. Auto-dissemination recently proved very efficient to control Aedes species, using adult females contaminated with dissemination stations of juvenile hormone to treat breeding habitats, but cannot be used at large scales. Here we propose to combine it to the Sterile Insect Technique (SIT) to create a new control concept, named 'boosted SIT' that might enable the area-wide eradication of mosquitoes and many other vectors and insect pests.

Contrasting patterns of tolerance between chemical and biological insecticides in mosquitoes exposed to UV-A.

Mosquitoes are vectors of major human diseases, such as malaria, dengue or yellow fever. Because no efficient treatments or vaccines are available for most of these diseases, control measures rely mainly on reducing mosquito populations by the use of insecticides. Numerous biotic and abiotic factors are known to modulate the efficacy of insecticides used in mosquito control. Mosquito breeding sites vary from opened to high vegetation covered areas leading to a large ultraviolet gradient exposure. This ecological feature may affect the general physiology of the insect, including the resistance status against insecticides. In the context of their contrasted breeding sites, we assessed the impact of low-energetic ultraviolet exposure on mosquito sensitivity to biological and chemical insecticides. We show that several mosquito detoxification enzyme activities (cytochrome P450, glutathione S-transferases, esterases) were increased upon low-energy UV-A exposure. Additionally, five specific genes encoding detoxification enzymes (CYP6BB2, CYP6Z7, CYP6Z8, GSTD4, and GSTE2) previously shown to be involved in resistance to chemical insecticides were found over-transcribed in UV-A exposed mosquitoes, revealed by RT-qPCR experiments. More importantly, toxicological bioassays revealed that UV-exposed mosquitoes were more tolerant to four main chemical insecticide classes (DDT, imidacloprid, permethrin, temephos), whereas the bioinsecticide Bacillus thuringiensis subsp. israelensis (Bti) appeared more toxic. The present article provides the first experimental evidence of the capacity of low-energy UV-A to increase mosquito tolerance to major chemical insecticides. This is also the first time that a metabolic resistance to chemical insecticides is linked to a higher susceptibility to a bioinsecticide. These results support the use of Bti as an efficient alternative to chemical insecticides when a metabolic resistance to chemicals has been developed by mosquitoes.

Genetic correlations and the evolution of photoperiodic time measurement within a local population of the pitcher-plant mosquito, Wyeomyia smithii.

The genetic relationship between the daily circadian clock and the seasonal photoperiodic timer remains a subject of intense controversy. In Wyeomyia smithii, the critical photoperiod (an overt expression of the photoperiodic timer) evolves independently of the rhythmic response to the Nanda-Hamner protocol (an overt expression of the daily circadian clock) over a wide geographical range in North America. Herein, we focus on these two processes within a single local population in which there is a negative genetic correlation between them. We show that antagonistic selection against this genetic correlation rapidly breaks it down and, in fact, reverses its sign, showing that the genetic correlation is due primarily to linkage and not to pleiotropy. This rapid reversal of the genetic correlation within a small, single population means that it is difficult to argue that circadian rhythmicity forms the necessary, causal basis for the adaptive divergence of photoperiodic time measurement within populations or for the evolution of photoperiodic time measurement among populations over a broad geographical gradient of seasonal selection.

Laboratory selection for an accelerated mosquito sexual development rate.

BACKGROUND: Separating males and females at the early adult stage did not ensure the virginity of females of Anopheles arabiensis (Dongola laboratory strain), whereas two years earlier this method had been successful. In most mosquito species, newly emerged males and females are not able to mate successfully. For anopheline species, a period of 24 h post-emergence is generally required for the completion of sexual maturation, which in males includes a 180° rotation of the genitalia. In this study, the possibility of an unusually shortened sexual maturity period in the laboratory-reared colony was investigated. METHODS: The effect of two different sex-separation methods on the virginity of females was tested: females separated as pupae or less than 16 h post-emergence were mated with males subjected to various doses of radiation. T-tests were performed to compare the two sex-separation methods. The rate of genitalia rotation was compared for laboratory-reared and wild males collected as pupae in Dongola, Sudan, and analysed by Z-tests. Spermatheca dissections were performed on females mated with laboratory-reared males to determine their insemination status. RESULTS: When the sex-separation was performed when adults were less than 16 h post-emergence, expected sterility was never reached for females mated with radio-sterilized males. Expected sterility was accomplished only when sexes were separated at the pupal stage. Observation of genitalia rotation showed that some males from the laboratory strain Dongola were able to successfully mate only 11 h after emergence and 42% of the males had already completed rotation. A small proportion of the same age females were inseminated. Wild males showed a much slower genitalia rotation rate. At 17 h post-emergence, 96% of the laboratory-reared males had completed genitalia rotation whereas none of the wild males had. CONCLUSION: This colony has been cultured in the laboratory for over one hundred generations, and now has accelerated sexual maturation when compared with the wild strain. This outcome demonstrates the kinds of selection that can be expected during insect colonization and maintenance, particularly when generations are non-overlapping and similar-age males must compete for mates.

Daily Plasmodium yoelii infective mosquito bites do not generate protection or suppress previous immunity against the liver stage.

BACKGROUND: Human populations that are naturally subjected to Plasmodium infection do not acquire complete protection against the liver stage of this parasite despite prolonged and frequent exposure. However, sterile immunity against Plasmodium liver stage can be achieved after repeated exposure to radiation attenuated sporozoites. The reasons for this different response remain largely unknown, but a suppressive effect of blood stage Plasmodium infection has been proposed as a cause for the lack of liver stage protection. METHODS: Using Plasmodium yoelii 17XNL, the response generated in mice subjected to daily infective bites from normal or irradiated mosquitoes was compared. The effect of daily-infected mosquito bites on mice that were previously immunized against P. yoelii liver stage was also studied. RESULTS: It was observed that while the bites of normal infected mosquitoes do not generate strong antibody responses and protection, the bites of irradiated mosquitoes result in high levels of anti-sporozoite antibodies and protection against liver stage Plasmodium infection. Exposure to daily infected mosquito bites did not eliminate the protection acquired previously with a experimental liver stage vaccine. CONCLUSIONS: Liver stage immunity generated by irradiated versus normal P. yoelii infected mosquitoes is essentially different, probably because of the blood stage infection that follows normal mosquito bites, but not irradiated. While infective mosquito bites do not induce a protective liver stage response, they also do not interfere with previously acquired liver stage protective responses, even if they induce a complete blood stage infection. Considering that the recently generated anti-malaria vaccines induce only partial protection against infection, it is encouraging that, at least in mouse models, immunity is not negatively affected by subsequent exposure and infection with the parasite.

SEM observation of the insect prepared by microwave irradiation.

The ptilinum of the fly and the compound eye are among the most fragile organs encountered during conventional procedures of morphological sample fixation. In order to identify a fixative suitable for preparing such samples for scanning electron microscopy, we examined various fixation conditions using microwave irradiation (MWI). The conditions examined were: (i) fixatives; (ii) temperature; (iii) concentration; (iv) duration; (v) dehydration; and (vi) substitution. The identified optimal conditions were 5% glutaraldehyde with MWI (350 W, 5 min). The MWI was continued until the maximal temperature of 75 degrees C was attained, followed by intermittent irradiation to maintain a temperature of 75 degrees C. After irradiation, the sample was left at room temperature for 24 h in the fixative and then dehydrated in increasing concentrations of ethanol. Each step in the ethanol series lasted for 24 h. The final absolute ethanol step included three solution changes, with each incubation lasting 1 h. A subsequent stepwise substitution of t-butyl alcohol for ethanol was conducted by reducing the ratio of 100% ethanol to t-butyl alcohol from 2:1 to 1:1 and then 1:2 (24 h each). The substitution was completed by three solution changes using 100% t-butyl alcohol, 30 min each. The best results were obtained by freeze-drying samples using t-butyl alcohol. The use of MWI improved fixative permeation, which occurred at a uniform rate throughout the sample. Comparison with temperature in a water bath at 75 degrees C indicated that the fixation effect of MWI was due to its heat generation in addition to some unknown mechanism.

Genetic shift in photoperiodic response correlated with global warming.

To date, all altered patterns of seasonal interactions observed in insects, birds, amphibians, and plants associated with global warming during the latter half of the 20th century are explicable as variable expressions of plastic phenotypes. Over the last 30 years, the genetically controlled photoperiodic response of the pitcher-plant mosquito, Wyeomyia smithii, has shifted toward shorter, more southern daylengths as growing seasons have become longer. This shift is detectable over a time interval as short as 5 years. Faster evolutionary response has occurred in northern populations where selection is stronger and genetic variation is greater than in southern populations. W. smithii represents an example of actual genetic differentiation of a seasonality trait that is consistent with an adaptive evolutionary response to recent global warming.