Keywords: Mosquito husbandry
Researchers develop temperature-controlled gene-editing method to potentially improve efforts to control disease-carrying insects
35356Caliann Ferguson, UT Health Houston School of Public Health, 2026-01-06 09:54:29.
New research presents promising results from an innovative technique that utilizes temperature control to genetically engineer sterile populations of insects, such as mosquitoes responsible for diseases like malaria, dengue, and other vector-borne illnesses. Led by researchers at UTHealth Houston School of Public Health, the Nature Communications publication leverages historical and traditional sterilization insect techniques (SIT) and applies an innovative method that can be scaled for larger population protection. Used for decades, traditional SITs include releasing large numbers of sterile males into mosquito populations so that when they mate with wild females, no viable offspring are produced. CRISPR/Cas9-based methods have proven challenging as they require breeding two separate lines of engineered insects and carefully sorting those insects to produce and release only sterile males. Led by principal investigator Victor Lopez Del Amo, PhD, assistant professor of epidemiology, and Christina Nguyen, a research technician, who carried out most of the experiments, this promising method aims to simplify the traditional SIT by harnessing a gene-editing tool called CRISPR-Cas12a that can generate male sterility and female lethality in a temperature-controlled manner. Cas12a can be engineered to be inactive at lower temperatures and active at higher temperatures. This property enabled the team to develop a single genetically modified insect strain that possesses the genetic composition necessary to disrupt key fertility and reproductive genes.
This scientist is breeding billions of mosquitoes to fight disease in Brazil
35301Mariana Lenharo, Nature, 2025-12-08 17:09:18.
nside a massive factory in the industrial district of Curitiba, Brazil, millions of Aedes aegypti mosquitoes are breeding in a climate-controlled room filled with mesh cages. Every week, the facility produces more than 80 million mosquito eggs. At the heart of this effort is Luciano Moreira, a soft-spoken agricultural engineer and entomologist, who opened the factory in July as part of an effort to fight mosquito-borne illnesses in the country. At the Curitiba facility, mosquitoes are infected with a bacterium called Wolbachia, which curbs the transmission of harmful human pathogens. Their offspring are being released in Brazilian cities to help to control dengue, a deadly viral disease transmitted mainly by A. aegypti. Until recently, Wolbachia-carrying mosquitoes were released only as part of small-scale research projects. The new factory marks a shift towards nationwide adoption of the method after Brazil’s federal government recognized it as an official public-health measure to combat dengue and other mosquito-borne diseases. People credit Moreira for making the case. “He has succeeded not only in carrying out the academic work, running experiments to demonstrate the model’s effectiveness, but also in convincing political decision-makers to implement the technology,” says Pedro Lagerblad de Oliveira, a molecular entomologist at Brazil’s Federal University of Rio de Janeiro. “This is a skill that not all scientists have.”
Female mosquito targeted with venom to curb disease
34263Dann Okoth, Scidev.net, 2025-01-14 10:10:57.
A genetic biocontrol method which reduces the lifespan of female insects could work as fast as pesticides to reduce populations of disease-spreading mosquitoes and destructive crop pests, according to researchers. Insect pests pose a huge threat to global health and agriculture, causing hundreds of thousands of deaths, millions of infections, and costing billions in healthcare and crop damage each year. Biocontrol is increasingly seen as a viable alternative to pesticides, which can harm non-target species and ecosystems and are losing efficacy as resistance to them grows. A new approach called Toxic Male Technique (TMT), developed by researchers at the ARC Centre of Excellence in Synthetic Biology at Macquarie University, Australia, involves genetically engineering male insects to reduce the lifespan of the females they mate with. Researchers say it can be used to respond rapidly to outbreaks of agricultural pests as well as to fight mosquito-borne diseases such as malaria, dengue and Zika. “We hold that our technology has the potential of working as fast as pesticides without the attendant risks of harming other species and the environment,” says Samuel Beach a researcher in applied biosciences at Macquarie University and lead author of the study, published today in Nature Communications. According to Beach, the approach is more efficient than existing methods such as the Sterile Insect Technique or the release of insects carrying lethal genes, which work by releasing masses of sterilized or genetically modified males to mate with wild females.
Genetically modified mosquitoes could combat deadly diseases, scientists say
34255The Sydney Morning Herald, 2025-01-13 15:25:27.
Genetically modified mosquitoes could be the solution to stopping deadly diseases spread by mosquito bites, scientists say.
Genetically Engineered Male Insects Shorten Their Mates’ Lifespans
34253Sneha Khedkar, The Scientist, 2025-01-13 15:13:56.
On a still night, as the air is thick with silence, the sharp, whining buzz of a mosquito shatters the calm. These blood-sucking insects that disturb people’s deep slumber are also responsible for spreading diseases such as dengue, chikungunya, malaria and Zika fever, which affect millions of people each year worldwide. Given the harmful effects of pesticides on the environment, combined with the emergence of mosquitoes resistant to pesticides, scientists are looking for alternative environment-friendly approaches for pest management. Now, researchers have developed a new population control method where male insects carrying toxic proteins can poison disease-spreading females during mating. The results, published in Nature Communications, describe a genetic biocontrol method that offers a fast and effective solution to managing pests. Such approaches are not entirely new. In the 1950s, when researchers mated female insects with radiologically sterilized males, they did not produce offsprings, reducing the next generation’s population. More recently, scientists propagated transgenes in insects that lower the fitness of future generations, resulting in decreased insect population. Although such methods are promising, they require at least one generation to take effect: Female insects may not produce offsprings, but they can continue transmitting infections. “As we’ve learned from COVID-19, reducing the spread of these diseases as quickly as possible is important to prevent epidemics,” said study author Samuel Beach, a graduate student in biologist Maciej Maselko’s lab at Macquarie University, in a press release.
Optimizing and synchronizing Aedes aegypti colony for Sterile Insect Technique application: Egg hatching, larval development, and adult emergence rate
31193Jennifer Thieme Castellon, Solomon Kibret Birhanie, Ale Macias, Rubi Casas, Jacob Hans, Michelle Q. Brown, Acta Tropica, 259. 2024-08-27 08:24:10.
Mosquito Sterile Insect Technique (SIT) programs can be developed in smaller agencies through synchronization of the colony development to take advantage of the natural male early emergence. This paper examined key aspects of Ae. aegypti colony synchronization work, including egg hatching, larval development, and adult emergence to produce sufficient numbers of adult male mosquitoes within a specific timeframe for irradiation and release. Our data indicated that a relatively low percentage of males are required for colony propagation. Additional results highlighted that fresher Ae. aegypti eggs could yield as high as a 93 % hatching success than older eggs when placed under vacuum pressure in yeast infused water for 1.5 h. Eggs that were one-month old hatched (93 %) better than older eggs (0–32 %). A higher egg density in the hatching flask was correlated to a lower hatch rate, and higher larval density was related to unsynchronized pupae and delayed adult emergence. By keeping Ae. aegypti larvae at reasonable density, over 95 % of adults emerged on the first two days of emergence – indicating a high synchronicity. A standardized colony maintenance protocol therefore renders a synchronized larval development and adult male emergence which are critical in SIT programs.
Effects of a blood-free mosquito diet on fitness and gonotrophic cycle parameters of laboratory reared Anopheles gambiae sensu stricto
30929Mosi, F.A., Rutha, I., Velez, R. et al., Parasites and Vectors, 17. 2024-07-24 15:43:32.
The current rise of new innovative tools for mosquito control, such as the release of transgenic mosquitoes carrying a dominant lethal gene and Wolbachia-based strategies, necessitates a massive production of mosquitoes in the insectary. However, currently laboratory rearing depends on vertebrate blood for egg production and maintenance. This practice raises ethical concerns, incurs logistical and cost limitations, and entails potential risk associated with pathogen transmission and blood storage. Consequently, an artificial blood-free diet emerges as a desirable alternative to address these challenges. This study aims to evaluate the effects of a previously formulated artificial blood-free diet (herein referred to as BLOODless) on Anopheles gambiae (An. gambiae s.s.; IFAKARA) gonotrophic parameters and fitness compared with bovine blood. The study was a laboratory-based comparative evaluation of the fitness, fecundity and fertility of An. gambiae s.s. (IFAKARA) reared on BLOODless versus vertebrate blood from founder generation (F0) to eighth generation (F8). A total of 1000 female mosquitoes were randomly selected from F0, of which 500 mosquitoes were fed with bovine blood (control group) and the other 500 mosquitoes were fed with BLOODless diet (experimental group). The feeding success, number of eggs per female, hatching rate and pupation rate were examined post-feeding. Longevity and wing length were determined as fitness parameters for adult male and female mosquitoes for both populations. While blood-fed and BLOODless-fed mosquitoes showed similar feeding success, 92.3% [95% confidence interval (CI) 89.7–94.9] versus 93.6% (95% CI 90.6–96.6), respectively, significant differences emerged in their reproductive parameters. The mean number of eggs laid per female was significantly higher for blood-fed mosquitoes (P < 0.001) whereas BLOODless-fed mosquitoes had significantly lower hatching rates [odds ratio (OR) 0.17, 95% CI 0.14–0.22, P < 0.001]. Wing length and longevity were similar between both groups. This study demonstrates the potential of the BLOODless diet as a viable and ethical alternative to vertebrate blood feeding for rearing An. gambiae s.s. This breakthrough paves the way for more efficient and ethical studies aimed at combating malaria and other mosquito-borne diseases.
Quantifying Fitness Costs in Transgenic Aedes aegypti Mosquitoes
27981I. Sanchez-Vargas, A. E. Williams, L. E. Martin, I. Martin-Martin, S. Bennett, K. E. Olson and E. Calvo, Journal of Visualized Experiments, 2023-09-15 07:51:42.
Transgenic mosquitoes often display fitness costs compared to their wild-type counterparts. In this regard, fitness cost studies involve collecting life parameter data from genetically modified mosquitoes and comparing them to mosquitoes lacking transgenes from the same genetic background. This manuscript illustrates how to measure common life history traits in the mosquito Aedes aegypti, including fecundity, wing size and shape, fertility, sex ratio, viability, development times, male contribution, and adult longevity. These parameters were chosen because they reflect reproductive success, are simple to measure, and are commonly reported in the literature. The representative results quantify fitness costs associated with either a gene knock-out or a single insertion of a gene drive element. Standardizing how life parameter data are collected is important because such data may be used to compare the health of transgenic mosquitoes generated across studies or to model the transgene fixation rate in a simulated wild-type mosquito population. Although this protocol is specific for transgenic Aedes aegypti, the protocol may also be used for other mosquito species or other experimental treatment conditions, with the caveat that certain biological contexts may require special adaptations.
How aggressive interactions with biomimetic agents optimize reproductive performances in mass-reared males of the Mediterranean fruit fly
25553D. Romano, G. Benelli and C. Stefanini, Biological Cybernetics, 2023-06-09 08:59:21.
Mass-rearing procedures of insect species, often used in biological control and Sterile Insect Technique, can reduce the insects competitiveness in foraging, dispersal, and mating. The evocation of certain behaviours responsible to induce specific neuroendocrine products may restore or improve the competitiveness of mass-reared individuals. Herein, we used a mass-reared strain of Ceratitis capitata as model organism. C. capitata is a polyphagous pest exhibiting territorial displays that are closely related to its reproductive performance. We tested if the behaviour of C. capitata males could be altered by hybrid aggressive interactions with a conspecific-mimicking robotic fly, leading to more competitive individuals in subsequent mating events. Aggressive interactions with the robotic fly had a notable effect on subsequent courtship and mating sequences of males that performed longer courtship displays compared to naïve individuals. Furthermore, previous interactions with the robotic fly produced a higher mating success of males. Reproductive performances of C. capitata males may be improved by specific octopaminergic neurones activated during previous aggressive interactions with the robotic fly. This study adds fundamental knowledge on the potential role of specific neuro-behavioural processes in the ecology of tephritid species and paves the way to innovative biotechnological control methods based on robotics and bionics.
From the Lab to the Field: Long-Distance Transport of Sterile Aedes Mosquitoes
24750H. Maïga, M. T. Bakhoum, W. Mamai, G. Diouf, N. S. Bimbilé Somda, T. Wallner, C. Martina, S. S. Kotla, O. B. Masso, H. Yamada, B. B. D. Sow, A. G. Fall and J. Bouyer, Insects, 14. 2023-02-18 19:03:39.
Pilot programs of the sterile insect technique (SIT) against Aedes aegypti may rely on importing significant and consistent numbers of high-quality sterile males from a distant mass rearing factory. As such, long-distance mass transport of sterile males may contribute to meet this requirement if their survival and quality are not compromised. This study therefore aimed to develop and assess a novel method for long-distance shipments of sterile male mosquitoes from the laboratory to the field. Different types of mosquito compaction boxes in addition to a simulation of the transport of marked and unmarked sterile males were assessed in terms of survival rates/recovery rates, flight ability and morphological damage to the mosquitoes. The novel mass transport protocol allowed long-distance shipments of sterile male mosquitoes for up to four days with a nonsignificant impact on survival (>90% for 48 h of transport and between 50 and 70% for 96 h depending on the type of mosquito compaction box), flight ability, and damage. In addition, a one-day recovery period for transported mosquitoes post-transport increased the escaping ability of sterile males by more than 20%. This novel system for the long-distance mass transport of mosquitoes may therefore be used to ship sterile males worldwide for journeys of two to four days. This study demonstrated that the protocol can be used for the standard mass transport of marked or unmarked chilled Aedes mosquitoes required for the SIT or other related genetic control programs.
Evaluating the mating competency of genetically modified male mosquitoes in laboratory conditions
28160B. Contreras, Z. N. Adelman and K. Chae, Frontiers in Tropical Diseases, 4. 2023-02-13 12:01:23.
Efforts to eradicate mosquito-borne diseases have increased the demand for genetic control strategies, many of which involve the release of genetically modified (GM) mosquito males into natural populations. The first hurdle for GM males is to compete with their wild-type counterparts for access to females. Here, we introduce an eye color-based mating assay, in which both Lvp wild-type and kynurenine 3-monooxygenase (kmo)-null males compete for access to kmo-null females, and therefore the eye color phenotype (black or white) of the progeny is dependent on the parental mating pair. A series of tests addressed that male mating competitiveness between the two strains can significantly be influenced by adult density, light intensity, and mating duration. Interestingly, the mating competitiveness of males was not correlated with body size, which was negatively influenced by a high larval density. Lastly, this eye color-associated assay was applied to characterize GM mosquitoes in their mating competitiveness, establishing this method as a fast and precise way of benchmarking this fitness parameter for laboratory-raised males.
Fitness costs of Wolbachia shift in locally-adapted Aedes aegypti mosquitoes
23703P. A. Ross and A. A. Hoffmann, Environmental Microbiology, 2022-10-06 08:10:07.
Aedes aegypti mosquito eggs can remain quiescent for many months before hatching, allowing populations to persist through unfavorable conditions. Aedes aegypti infected with the Wolbachia strain wMel have been released in tropical and subtropical regions for dengue control. wMel reduces the viability of quiescent eggs, but this physiological cost might be expected to evolve in natural mosquito populations that frequently experience stressful conditions. We found that the cost of wMel infection differed consistently between mosquitoes collected from different locations and became weaker across laboratory generations, suggesting environment-specific adaptation of mosquitoes to the wMel infection. Reciprocal crossing experiments show that differences in the cost of wMel to quiescent egg viability were mainly due to mosquito genetic background and not Wolbachia origin. wMel-infected mosquitoes hatching from long-term quiescent eggs showed partial loss of cytoplasmic incompatibility and female infertility, highlighting additional costs of long-term quiescence. Our study provides the first evidence for a shift in Wolbachia phenotypic effects following deliberate field release and establishment and it highlights interactions between Wolbachia infections and mosquito genetic backgrounds. The unexpected changes in fitness costs observed here suggest potential tradeoffs with undescribed fitness benefits of the wMel infection.
Fitness effects of CRISPR endonucleases in Drosophila melanogaster populations
23673A. M. Langmüller, J. Champer, S. Lapinska, L. Xie, M. Metzloff, S. E. Champer, J. Liu, Y. Xu, J. Du, A. G. Clark and P. W. Messer, eLife, 11:e71809. 2022-09-22 08:22:04.
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 provides a highly efficient and flexible genome editing technology with numerous potential applications ranging from gene therapy to population control. Some proposed applications involve the integration of CRISPR/Cas9 endonucleases into an organism’s genome, which raises questions about potentially harmful effects to the transgenic individuals. One example for which this is particularly relevant are CRISPR-based gene drives conceived for the genetic alteration of entire populations. The performance of such drives can strongly depend on fitness costs experienced by drive carriers, yet relatively little is known about the magnitude and causes of these costs. Here, we assess the fitness effects of genomic CRISPR/Cas9 expression in Drosophila melanogaster cage populations by tracking allele frequencies of four different transgenic constructs that allow us to disentangle ‘direct’ fitness costs due to the integration, expression, and target-site activity of Cas9, from fitness costs due to potential off-target cleavage. Using a maximum likelihood framework, we find that a model with no direct fitness costs but moderate costs due to off-target effects fits our cage data best. Consistent with this, we do not observe fitness costs for a construct with Cas9HF1, a high-fidelity version of Cas9. We further demonstrate that using Cas9HF1 instead of standard Cas9 in a homing drive achieves similar drive conversion efficiency. These results suggest that gene drives should be designed with high-fidelity endonucleases and may have implications for other applications that involve genomic integration of CRISPR endonucleases.
Local adaptation of Aedes aegypti mosquitoes to Wolbachia-induced fitness costs
22208P. A. Ross and A. A. Hoffmann, bioRxiv, 2022.05.06.490959. 2022-05-06 15:19:41.
Aedes aegypti mosquito eggs can remain quiescent for many months before hatching, allowing populations to persist through unfavorable conditions. Aedes aegypti infected with the Wolbachia strain wMel have been released in tropical and subtropical regions for dengue control. wMel reduces the viability of quiescent eggs, but this physiological cost might be expected to evolve in natural mosquito populations that frequently experience stressful conditions. We therefore compared the costs of wMel infection for quiescent egg viability in field-derived and laboratory populations. Quiescent egg viability was highly variable in wMel-infected populations, with greater costs of wMel in field-derived populations. In contrast, there was little variation between matched field-derived and long-term laboratory populations lacking wMel, suggesting that laboratory adaptation does not influence this trait and that differences are due to wMel infection. Comparisons of populations collected a year apart show a decline in costs under laboratory rearing conditions involving a rapid turnover of mosquito generations; this pattern was consistent across populations despite their origin, suggesting adaptation of mosquitoes to the wMel infection under laboratory conditions. Reciprocal crossing experiments confirm that differences in quiescent egg viability were mainly due to the genetic background and not Wolbachia alone. wMel-infected mosquitoes hatching from long-term quiescent eggs showed partial loss of cytoplasmic incompatibility and female infertility, highlighting additional costs of long-term quiescence. Our study provides the first evidence for a shift in Wolbachia phenotypic effects following deliberate field release and establishment and it highlights interactions between Wolbachia infections and local adaptation. The unexpected changes in fitness costs observed here suggest potential tradeoffs with undescribed fitness benefits of the wMel infection.Competing Interest StatementThe authors have declared no competing interest.
Pilot trial using mass field-releases of sterile males produced with the incompatible and sterile insect techniques as part of integrated Aedes aegypti control in Mexico
22224A. Martín-Park, A. Che-Mendoza, Y. Contreras-Perera, S. Pérez-Carrillo, H. Puerta-Guardo, J. Villegas-Chim, G. Guillermo-May, A. Medina-Barreiro, H. Delfín-González, R. Méndez-Vales, S. Vázquez-Narvaez, J. Palacio-Vargas, F. Correa-Morales, G. Ayora-Tal, PLoS Negl Trop Dis, 16:e0010324. 2022-04-26 15:46:43.
We implemented a controlled before-and-after quasi-experimental study in two suburban localities of Yucatan (Mexico): San Pedro Chimay (SPC), which received IIT-SIT, and San Antonio Tahdzibichén used as control. Release of wAlbB Ae. aegypti males at SPC extended for 6 months (July-December 2019), covering the period of higher Ae. aegypti abundance. Entomological indicators included egg hatching rates and outdoor/indoor adult females collected at the release and control sites. Approximately 1,270,000 lab-produced wAlbB-infected Ae. aegypti males were released in the 50-ha treatment area (2,000 wAlbB Ae. aegypti males per hectare twice a week in two different release days, totaling 200,000 male mosquitoes per week). The efficacy of IIT-SIT in suppressing indoor female Ae. aegypti density (quantified from a generalized linear mixed model showing a statistically significant reduction in treatment versus control areas) was 90.9% a month after initiation of the suppression phase, 47.7% two months after (when number of released males was reduced in 50% to match local abundance), 61.4% four months after (when initial number of released males was re-established), 88.4% five months after and 89.4% at six months after the initiation of the suppression phase. A proportional, but lower, reduction in outdoor female Ae. aegypti was also quantified (range, 50.0-75.2% suppression). CONCLUSIONS/SIGNIFICANCE: Our study, the first open-field pilot implementation of Wolbachia IIT-SIT in Mexico and Latin-America, confirms that inundative male releases can significantly reduce natural populations of Ae. aegypti. More importantly, we present successful pilot results of the integration of Wolbachia IIT-SIT within a IVM plan implemented by Ministry of Health personnel.
Mark-release-recapture experiment in Burkina Faso demonstrates reduced fitness and dispersal of genetically-modified sterile malaria mosquitoes
20385F. A. Yao, A.-A. Millogo, P. S. Epopa, A. North, F. Noulin, K. Dao, M. Drabo, C. Guissou, S. Kekele, M. Namountougou, R. K. Ouedraogo, L. Pare, N. Barry, R. Sanou, H. Wandaogo, R. K. Dabire, A. McKemey, F. Tripet and A. Diabaté, Nature Communications, 13:796. 2022-02-10 09:11:33.
Every year, malaria kills approximately 405,000 people in Sub-Saharan Africa, most of them children under the age of five years. In many countries, progress in malaria control has been threatened by the rapid spread of resistance to antimalarial drugs and insecticides. Novel genetic mosquito control approaches could play an important role in future integrated malaria control strategies. In July 2019, the Target Malaria consortium proceeded with the first release of hemizygous genetically-modified (GM) sterile and non-transgenic sibling males of the malaria mosquito Anopheles coluzzii in Burkina Faso. This study aimed to determine the potential fitness cost associated to the transgene and gather important information related to the dynamic of transgene-carrying mosquitoes, crucial for next development steps. Bayesian estimations confirmed that GM males had lower survival and were less mobile than their wild type (WT) siblings. The estimated male population size in Bana village, at the time of the release was 28,000 - 37,000. These results provide unique information about the fitness and behaviour of released GM males that will inform future releases of more effective strains of the A. gambiae complex.
Quality Control Methods for Aedes albopictus Sterile Male Transportation
20392G. D. Mastronikolos, A. Kapranas, G. K. Balatsos, C. Ioannou, D. P. Papachristos, P. G. Milonas, A. Puggioli, I. Pajović, D. Petrić, R. Bellini, A. Michaelakis and N. T. Papadopoulos, Insects, 2022-02-09 09:27:23.
Genetic based mosquito control methods have been gaining ground in recent years for their potential to achieve effective suppression or replacement of vector populations without hampering environments or causing any public health risk. These methods require the mass rearing of the target species in large facilities sized to produce millions of sterile males, as already well established for a number of insects of agricultural importance. Assessing the performance of released males in Sterile Insect Technique (SIT) control programs is of the utmost importance for the success of the operation. Besides the negative effects of mass rearing and sterilization, the handling of sterilized insects and shipment to distant areas may also negatively impact the quality of sterilized males. The aim of the current study was to design and executive quality control (QC) tests for sterilized Aedes albopictus (Asian tiger mosquito) males delivered by air shipment from the mass production facility located in Italy to Greece and Montenegro field release sites. Mass reared mosquito strains were based on biological materials received from Italy, Greece and Montenegro. Tests conducted at the mass rearing facility before transportation revealed a rather high residual female contamination following mechanical sex separation (approximately 1.5% females, regardless of the mosquito strain). Irradiated males of all three mosquito strains induced high levels of sterility to females. Shipment lasting approximately 24 h resulted in approximately 15% mortality, while when shipment lasted nearly two days this increased to almost 40%. The flight ability of sterilized males following one day transportation time was satisfactory (over 60%). The response of sterile males to food and water starvation was comparable and slightly lower than that of wild non-transported males. Longevity of sterile males was shorter than that of wild counterparts and it seems it was not affected by mating to wild females. Both mating propensity and mating competitiveness for wild virgin females was higher for the wild, control males compared to the sterile, transported ones. Overall, the performance of sterile male Ae. albopictus delivered from the mass rearing facility of Italy to Greece in approximately 24 h was satisfactory. Transportation lasting two days or longer incurred detrimental effects on males, which called into question the outcome of the SIT release programs. In conclusion, our results demonstrate the need of quality control procedures, especially when sterile male production facilities are not near to the releasing point. Transportation could be a serious drawback for the implementation of Sterile Insect Releases and, consequently, it is important to establish an efficient and fast transportation of sterilized males in advance.
Lab-scale characterization and semi-field trials of Wolbachia Strain wAlbB in a Taiwan Wolbachia introgressed Ae. aegypti strain
20383W. L. Liu, H. Y. Yu, Y. X. Chen, B. Y. Chen, S. N. Leaw, C. H. Lin, M. P. Su, L. S. Tsai, Y. Chen, S. H. Shiao, Z. Y. Xi, A. C. C. Jang and C. H. Chen, PLOS Neglected Tropical Diseases, 16:24. 2022-01-11 08:53:33.
Author summaryPrior to open field release, new genetic approaches that interfere with mosquito abilities and reduce mosquito population density require progressive evaluation both in the laboratory and contained field trials. Trials in contained outdoor systems are thus an important step and allow for evaluation of the effectiveness and reliability of Wolbachia-infected Aedes aegypti (wAlbB-Tw) in settings that incorporate natural environmental variations. We describe a semi-field system designed to study the ability of wAlbB-Tw mosquitoes to suppress local mosquito populations. We provide a protocol for establishing, maintaining, and monitoring Ae. aegypti population densities inside field cages. These results pave the way for the open release of wAlbB-Tw-infected Ae. aegypti mosquitoes in the field. Dengue fever is one of the most severe viral diseases transmitted by Aedes mosquitoes, with traditional approaches of disease control proving insufficient to prevent significant disease burden. Release of Wolbachia-transinfected mosquitoes offers a promising alternative control methodologies; Wolbachia-transinfected female Aedes aegypti demonstrate reduced dengue virus transmission, whilst Wolbachia-transinfected males cause zygotic lethality when crossed with uninfected females, providing a method for suppressing mosquito populations. Although highly promising, the delicate nature of population control strategies and differences between local species populations means that controlled releases of Wolbachia-transinfected mosquitoes cannot be performed without extensive testing on specific local Ae. aegypti populations. In order to investigate the potential for using Wolbachia to suppress local Ae. aegypti populations in Taiwan, we performed lab-based and semi-field fitness trials. We first transinfected the Wolbachia strain wAlbB into a local Ae. aegypti population (wAlbB-Tw) and found no significant changes in lifespan, fecundity and fertility when compared to controls. In the laboratory, we found that as the proportion of released male mosquitoes carrying Wolbachia was increased, population suppression could reach up to 100%. Equivalent experiments in semi-field experiments found suppression rates of up to 70%. The release of different ratios of wAlbB-Tw males in the semi-field system provided an estimate of the optimal size of male releases. Our results indicate that wAlbB-Tw has significant potential for use in vector control strategies aimed at Ae. aegypti population suppression in Taiwan. Open field release trials are now necessary to confirm that wAlbB-Tw mediated suppression is feasible in natural environments.
Beyond the eye: Kynurenine pathway impairment causes midgut homeostasis dysfunction and survival and reproductive costs in blood-feeding mosquitoes
19915V. Bottino-Rojas, I. Ferreira, R. D. Nunes, X. Feng, T. B. Pham, A. Kelsey, R. Carballar-Lejarazú, V. Gantz, P. L. Oliveira and A. A. James, Insect Biochemistry and Molecular Biology, 103720. 2022-01-06 08:57:05.
Insect ommochrome biosynthesis pathways metabolize tryptophan to generate eye-color pigments and naturally occurring alleles of pathway genes are useful phenotypic markers in transgenesis studies. Pleiotropic effects of mutations in some genes exert a load on both survival and reproductive success in blood-feeding species. Here, we investigated the challenges imposed on mosquitoes by the increase of tryptophan resulting from blood meal digestion and the impact of disruptions of the ommochrome biosynthesis pathway. Female mosquitoes with spontaneous and induced mutations in the orthologs of the genes encoding kynurenine hydroxylase in Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus exhibited impaired survival and reproductive phenotypes that varied in type and severity among the species. A compromised midgut permeability barrier function was also observed in An. stephensi. Surprisingly, mutant mosquitoes displayed an increase in microbiota compared to controls that was not accompanied by a general induction of immune genes. Antibiotic treatment rescued some deleterious traits implicating a role for the kynurenine pathway (KP) in midgut homeostasis. Supplemental xanthurenic acid, a KP end-product, rescued lethality and limited microbiota proliferation in Ae. aegypti. These data implicate the KP in the regulation of the host/microbiota interface. These pleiotropic effects on mosquito physiology are important in the development of genetic strategies targeting vector mosquitoes.
Temporal Viability of Aedes aegypti and Aedes albopictus Eggs Using Two Hygroscopic Substances as Preservatives under a Sterile Insect Technique (SIT) Program in Southern Mexico
19721E. N. Martínez-García, E. E. Díaz-González, C. F. Marina, J. G. Bond, J. J. Rodríguez-Rojas, G. Ponce-García, R. M. Sánchez-Casas and I. Fernández-Salas, Insects, 13. 2021-12-21 12:31:48.
Dengue and other Aedes-borne diseases have dramatically increased over the last decades. The Sterile Insect Technique (SIT) has been successfully used as part of integrated pest strategies to control populations of insect-plant and livestock pests and is currently being tested as a potential method to reduce mosquito populations in an environmentally friendly approach. However, during the mass rearing steps needed to produce millions of mosquitoes, egg storage and preservation are essential for a certain amount of time. Eggs of Aedes aegypti have a chorionic pad that functions as a sticky substance to glue them onto the inner walls of larval breeding sites. The chorionic pad is chemically made of hyaluronic acid, a hygroscopic compound, responsible to protect them from desiccation over time. Two commercial products with hygroscopic properties, hydrolyzed collagen, and Hyalurosmooth®, both were tested to assess their ability to prolong egg life storage for A. aegypti and A. albopictus. Results showed that 85–95% of Ae. aegypti eggs were able to hatch up to week 8 after being treated with both hydrophilic compounds, compared with the control 66.3%. These two substances showed promising effects for keeping Ae. aegypti eggs viable during prolonged storage in mass rearing insect production focused on vector control SIT programs.
The Possible Role of Microorganisms in Mosquito Mass Rearing
17741L. Chersoni, A. Checcucci, M. Malfacini, A. Puggioli, F. Balestrino, M. Carrieri, I. Piunti, M. L. Dindo, P. Mattarelli and R. Bellini, Insects, 12. 2021-07-15 13:10:37.
In Europe, one of the most significant mosquitoes of public health importance is Aedes albopictus (Skuse), an allochthonous species of Asian origin. One of the most promising control methods against Aedes albopictus is the sterile insect technique (SIT), which consists of mass rearing the target species, separation of males from females, and male exposure to sterilizing ionizing radiation. Once released in the environment, the sterile males are expected to search for wild females to mate with. If mating occurs, no offspring is produced. The quality of sterile males is a crucial aspect in SIT programs in order to optimize effectiveness and limit production costs. The integration of probiotic microorganisms in larval and adult mosquito diets could enhance the quality parameters of the released sterile males. In this review, we attempt to give the most representative picture of the present knowledge on the relationships between gut microbiota of mosquitoes and the natural or artificial larval diet. Furthermore, the possible use of probiotic microorganisms for mosquito larvae rearing is explored. Based on the limited amount of data found in the literature, we hypothesize that a better understanding of the interaction between mosquitoes and their microbiota may bring significant improvements in mosquito mass rearing for SIT purposes.
Effect of the timing of pupal irradiation on the quality and sterility of oriental fruit flies (Diptera: Tephritidae) for use in Sterile Insect Technique
17781T. J. Fezza, P. A. Follett and T. E. Shelly, Applied Entomology and Zoology, 8. 2021-07-05 17:37:25.
The Sterile Insect Technique (SIT) is a target-specific, biologically based method used to control pestiferous tephritids entailing the release of mass-reared, sterilized males of the target species to achieve sterile male x wild female matings. As documented for several tephritid species, sterilizing irradiation may have adverse effects on various biological parameters, including life span, flight ability, and mating competitiveness. To minimize these impacts and ensure sterility, released flies must be irradiated at a precise dose at a specific and uniform age. The objective of the present study was to determine the impact of pupal age at the time of irradiation on flight ability, male survival, fertility, and mating competitiveness for the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) irradiated at 100 Gy. We concluded that flies irradiated as pupae 4 days before emergence were significantly compromised with respect to flight ability and male survival when compared to flies irradiated 1 or 2 days before emergence. Additionally, we determined that crosses between males irradiated 1 day before emergence with non-irradiated females yielded significantly more pupae than the same cross with males irradiated at 2, 3, and 4 days before emergence. Although rare, the occurrence of pupae and emerged adults suggested low levels of fertility for flies irradiated as pupae 1 day before emergence, but complete sterility at 2, 3, and 4 days before emergence. These findings suggest that SIT programs can potentially irradiate pupae at 2 or 3 days before emergence without compromising the quality of the released flies, which would permit the distribution of flies over greater distances.
Evaluating Flight Performance of Mass-Reared and Irradiated Navel Orangeworm (Lepidoptera: Pyralidae) for Sterile Insect Technique
17545J. Reger, J. A. Wenger, G. Brar, C. Burks and H. Wilson, Journal of Economic Entomology, 2021-06-21 13:28:59.
Navel orangeworm (Pyralidae: Amyelois transitella) is a key pest of almonds and pistachios in California. Moths directly infest nuts which leads to reduced crop yield and quality, and infestation can predispose nuts to fungal pathogens that produce aflatoxins. While several integrated pest management strategies have been developed for A. transitella, studies have recently been initiated to explore the use of sterile insect technique (SIT) as an additional control tool. Mass-rearing, sterilization, and transportation methods originally developed for Pectinophora gossypiella (Lepidoptera: Gelechiidae) are currently being used for production of A. transitella in a mass-rearing facility, but the impacts of these processes on performance of A. transitella remain unclear. In this study, computerized flight mills were used to evaluate multiple flight parameters of mass-reared and irradiated A. transitella males and females relative to non-irradiated mass-reared moths and two strains of locally reared moths which were neither mass-reared nor irradiated. Mass-reared non-irradiated females performed similarly to both strains of locally reared females, flying a mean 9.4–11.8 km per night, whereas mass-reared and irradiated males and mass-reared non-irradiated males all flew shorter distances, in the range of 3.0–6.7 km per night. All of the mass-reared moths compared to locally reared moths had significantly more non-fliers that did not engage in more than two minutes of continuous flight. Findings from this study suggest that mass-rearing conditions reduce A. transitella flight capacity, while irradiation interacts with moths in a sex-specific manner.
Development of Sterile Insect Technique for Control of the European Grapevine Moth, Lobesia botrana, in Urban Areas of Chile
16849G. S. Simmons, M. C. Salazar Sepulveda, E. A. Fuentes Barrios, M. Idalsoaga Villegas, R. E. Medina Jimenez, A. R. Garrido Jerez, R. Henderson and H. Donoso Riffo, Insects, 12. 2021-04-22 13:20:22.
The European grapevine moth, a Palearctic pest, was first detected in the Americas in 2008. Its establishment in Chile presented production and export issues for grapes and other fruits, and a national control campaign was launched. Urban areas next to agricultural production areas were recognized as a challenge for effective control. In 2015, a SIT laboratory was established in Arica, Chile to evaluate its potential for urban control. Progress included the development and evaluation of artificial diets, a mass-rearing of 75,000 moths/week, confirmation of 150 Gy as an operational dose for inherited sterility, and releases of sterile moths in a 25 ha urban area next to fruit production areas. Season-long releases demonstrated that high overflooding ratios were achieved early in the season but decreased with a large increase in the wild moth population. Sterile moth quality was consistently high, and moths were observed living in the field up to 10 days and dispersing up to 800 m. Recommendations for further development of the SIT include conducting cage and field studies to evaluate overflooding ratios and mating competitiveness, measuring of infestation densities in release and no-release areas, and conducting trials to evaluate combining SIT with compatible integrated pest management (IPM) tactics such as fruit stripping and use of mating disruption.
The Insect Pest Control Laboratory of the Joint FAO/IAEA Programme: Ten Years (2010–2020) of Research and Development, Achievements and Challenges in Support of the Sterile Insect Technique
16702M. J. B. Vreysen, A. M. M. Abd-Alla, K. Bourtzis, J. Bouyer, C. Caceres, C. de Beer, D. Oliveira Carvalho, H. Maiga, W. Mamai, K. Nikolouli, H. Yamada and R. Pereira, Insects, 12. 2021-04-13 15:29:17.
The Joint FAO/IAEA Centre (formerly called Division) of Nuclear Techniques in Food and Agriculture was established in 1964 and its accompanying laboratories in 1961. One of its subprograms deals with insect pest control, and has the mandate to develop and implement the sterile insect technique (SIT) for selected key insect pests, with the goal of reducing the use of insecticides, reducing animal and crop losses, protecting the environment, facilitating international trade in agricultural commodities and improving human health.
Sterile Insect Technique in an Integrated Vector Management Program against Tiger Mosquito Aedes albopictus in the Valencia Region (Spain): Operating Procedures and Quality Control Parameters
16663C. Tur, D. Almenar, S. Benlloch-Navarro, R. Argilés-Herrero, M. Zacarés, V. Dalmau and I. Pla, Insects, 12. 2021-03-23 14:20:04.
The Agriculture Department of the Valencian Region is promoting an ongoing pilot project to evaluate the efficacy of an integrated vector management program (IVM) based on the use of the SIT as the main method of control. The laboratory studies for evaluating the entomological efficacy of SIT through the phased conditional testing process recommended by World Health Organization and the International Atomic Energy Agency (WHO-IAEA) are addressed. This study describes the routine operating procedures and quality control parameters for the medium-scale rearing of sterile male A. albopictus. More than 15 million sterile males have been produced and released in an area of 80 ha between 2018 and 2020. Of the initial L1 larvae, we recovered 17.2% of male pupae after sex sorting to be sterilized and released on the field, while the rest of the pupae remained available to maintain the rearing colony. The residual percentage of females after sex sorting was on average 0.17%. The obtained values in terms of production and quality control as well as the proposed rearing methodology can be useful for designing a medium-scale mosquito-rearing pipeline.
Improving the Phenotypic Properties of the Ceratitis capitata (Diptera: Tephritidae) Temperature-Sensitive Lethal Genetic Sexing Strain in Support of Sterile Insect Technique Applications
16226M. F. Porras, J. S. Meza, E. G. Rajotte, K. Bourtzis and C. Caceres, Journal of Economic Entomology, 113:2688-2694. 2021-01-31 16:33:01.
We present the first evidence that this slower development is due to a different gene, here namely slow development (sd), which is closely linked to the tsl gene. Taking advantage of recombination phenomena between the two loci, we report the isolation of a novel temperature sensitivity lethal strain using the wp mutation as a morphological marker, which showed faster development (wp tsl FD) during the larval stage and increased in its temperature sensitivity compared with the normal tsl strain. Moreover, the introgression of this novel wp tsl FD combined trait into the Vienna 8(D53-) GSS, resulted in a novel Vienna 8(D53-) FD GSS, where females showed differences in the thermal sensibility, larval development speed, and productivity profiles. The modification of these traits and their impact on the mass rearing of the GSS for sterile insect technique applications are discussed.
Twenty-five Years of Research Experience with the Sterile Insect Technique and Area-Wide Management of Codling Moth, Cydia pomonella (L.), in Canada
13154H. M. A. Thistlewood and G. J. R. Judd, Insects, 10:292. 2019-09-10 14:21:45.
The advent of novel genetic methods has led to renewed interest in the sterile insect technique (SIT) for management of insect pests, owing to applications in mass rearing and in the production of sterile offspring without use of irradiation. An area-wide management programme for codling moth, Cydia pomonella, has employed the SIT and other management practices over a large area (3395 to 7331 ha) of orchards and neighbouring urban, public, or First Nations lands in British Columbia, Canada, for 25 years. This project is the first to employ the SIT for C. pomonella, and the longest-running application of area-wide techniques for its control, anywhere. It was derived from basic research and applied trials from the 1960s onwards. Many biological challenges were overcome, and lessons learnt, in transferring from small- to large-scale applications of mass rearing and the SIT, with particular regard to Lepidoptera. Research has proven essential to identifying, if not resolving, issues that threaten the implementation and success of any such programme. The major challenges encountered, and the resulting research, are reviewed, as well as future directions. Recommendations are given for application of the SIT as part of any area-wide management programme for C. pomonella.
Gene-drive-mediated extinction is thwarted by population structure and evolution of sib mating
11202J. J. Bull, C. H. Remien and S. M. Krone, Evolution Medicine and Public Health, 2019:66-81. 2019-05-11 14:03:57.
Genetic engineering combined with CRISPR technology has developed to the point that gene drives can, in theory, be engineered to cause extinction in countless species. Success of extinction programs now rests on the possibility of resistance evolution, which is largely unknown. Depending on the gene-drive technology, resistance may take many forms, from mutations in the nuclease target sequence (e.g. for CRISPR) to specific types of non-random population structures that limit the drive (that may block potentially any gene-drive technology).
Controlling invasive rodents via synthetic gene drive and the role of polyandry
3927Manser, AC, S. J.; Sutter, A.; Blondel, D. V.; Serr, M.; Godwin, J.; Price, T. A. R., Proceedings of the Royal Society B-Biological Sciences, 286:9. 2019-01-05 00:00:00.
House mice are a major ecosystem pest, particularly threatening island ecosystems as a non-native invasive species. Rapid advances in synthetic biology offer new avenues to control pest species for biodiversity conservation. Recently, a synthetic sperm-killing gene drive construct called t-Sry has been proposed as a means to eradicate target mouse populations owing to a lack of females. A factor that has received little attention in the discussion surrounding such drive applications is polyandry. Previous research has demonstrated that sperm-killing drivers are extremely damaging to a male's sperm competitive ability. Here, we examine the importance of this effect on the t-Sry system using a theoretical model. We find that polyandry substantially hampers the spread of t-Sry such that release efforts have to be increased three-to sixfold for successful eradication. We discuss the implications of our finding for potential pest control programmes, the risk of drive spread beyond the target population, and the emergence of drive resistance. Our work highlights that a solid understanding of the forces that determine drive dynamics in a natural setting is key for successful drive application, and that exploring the natural diversity of gene drives may inform effective gene drive design.
The Trojan Female Technique for pest control: a candidate mitochondrial mutation confers low male fertility across diverse nuclear backgrounds in Drosophila melanogaster
13733D. K. Dowling, D. M. Tompkins and N. J. Gemmell, Evolutionary Applications, 8:8710880. 2015-07-15 19:28:56.
The Trojan Female Technique (TFT) was recently proposed as a prospective approach to biological pest control. However, applicability of the TFT relies on mitochondrial mutations whose male-sterilizing effects are general across nuclear genomic contexts. We test this assumption, expressing the candidate TFT-mutation bearing haplotype alongside a range of nuclear backgrounds and comparing its fertility in males, relative to that of control haplotypes
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