scholarly journals Using the Fluorescent Dye, Rhodamine B, to Study Mating Competitiveness in Male Aedes aegypti Mosquitoes

10.3791/62432 ◽  
2021 ◽  
Author(s):  
Irene Li ◽  
Keng Wai Mak ◽  
Jeslyn Wong ◽  
Cheong Huat Tan
Keyword(s):  
Aedes Aegypti ◽  
Rhodamine B ◽  
Fluorescent Dye ◽  
Mating Competitiveness

scholarly journals Fluorescent markers rhodamine B and uranine for Anopheles gambiae adults and matings

2020 ◽  
Author(s):  
Erica i Aviles ◽  
Rachel D Rotenberry ◽  
C Matilda Collins ◽  
Ellen M Dotson ◽  
Mark Q Benedict
Keyword(s):  
Anopheles Gambiae ◽  
Rhodamine B ◽  
Seminal Fluid ◽  
Adult Longevity ◽  
Male Mating ◽  
Potential Contribution ◽  
Laboratory Studies ◽  
Mating Competitiveness ◽  
Fluorescent Markers ◽  
Auto Fluorescence

Abstract Background Marking mosquitoes is vital for mark-release-recapture and many laboratory studies, but their small size precludes the use of methods that are available for larger animals such as unique identifier tags and radio devices. Fluorescent dust is the most commonly used method to distinguish released individuals from the wild population. Numerous colours and combinations can be used, however, dust sometimes affects longevity and behaviour so alternatives that do not have these effects would contribute substantially. Rhodamine B has previously been demonstrated to be useful for marking adult Aedes aegypti males when added to the sugar meal. Unlike dust, this also marked the seminal fluid making it possible to detect matings by marked males in the spermatheca of females. Here, marking of Anopheles gambiae sensu stricto with rhodamine B and uranine was performed to estimate their potential contribution. Methods Two fluorescent markers, rhodamine B and uranine, were dissolved in sugar water and fed to adult An. gambiae. Concentrations that are useful for marking individuals and seminal fluid were determined. The effects on adult longevity, the durability of the marking and detection of the marker in mated females was determined. Male mating competitiveness was also evaluated.Results Rhodamine B marking in adults is detectable for at least three weeks, however uranine marking declines with time and at low doses can be confused with auto-fluorescence. Both can be used for marking seminal fluid which can be detected in females mated by marked males, but, again, at low concentrations uranine-marking is more easily confused with the natural fluorescence of seminal fluid. Neither dye affected mating competitiveness.Conclusions Both markers tested could be useful for field and laboratory studies. Their use has substantial potential to contribute to a greater understanding of the bio-ecology of this important malaria vector. Rhodamine B has the advantage that it appears to be permanent and is less easily confused with auto-fluorescence. The primary limitation of both methods is that sugar feeding is necessary for marking and adults must be held for at least 2 nights to ensure all individuals are marked whereas dusts provide immediate and thorough marking.

scholarly journals Mating competitiveness and life‐table comparisons between transgenic and Indian wild‐type Aedes aegypti L.

2014 ◽  
Vol 71 (7) ◽  
pp. 957-965 ◽  
Author(s):  
Prabhakargouda B Patil ◽  
BP Niranjan Reddy ◽  
Kevin Gorman ◽  
KV Seshu Reddy ◽  
Shirish R Barwale ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Life Table ◽  
Wild Type ◽  
Mating Competitiveness

scholarly journals Mark-release-recapture of male Aedes aegypti (Diptera: Culicidae): Use of rhodamine B to estimate movement, mating and population parameters in preparation for an incompatible male program

2021 ◽  
Vol 15 (6) ◽  
pp. e0009357
Author(s):  
Brendan J. Trewin ◽  
Daniel E. Pagendam ◽  
Brian J. Johnson ◽  
Chris Paton ◽  
Nigel Snoad ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Rhodamine B ◽  
Population Control ◽  
Invasive Disease ◽  
Population Surveillance ◽  
Disease Vectors ◽  
Male Population ◽  
Species Survival ◽  
Mark Release Recapture ◽  
Wide Population

Rapid advances in biological and digital support systems are revolutionizing the population control of invasive disease vectors such as Aedes aegypti. Methods such as the sterile and incompatible insect techniques (SIT/IIT) rely on modified males to seek out and successfully mate with females, and in doing so outcompete the wild male population for mates. Currently, these interventions most frequently infer mating success through area-wide population surveillance and estimates of mating competitiveness are rare. Furthermore, little is known about male Ae. aegypti behaviour and biology in field settings. In preparation for a large, community scale IIT program, we undertook a series of mark- release-recapture experiments using rhodamine B to mark male Ae. aegypti sperm and measure mating interactions with females. We also developed a Spatial and Temporally Evolving Isotropic Kernel (STEIK) framework to assist researchers to estimate the movement of individuals through space and time. Results showed that ~40% of wild females captured daily were unmated, suggesting interventions will need to release males multiple times per week to be effective at suppressing Ae. aegypti populations. Males moved rapidly through the landscape, particularly when released during the night. Although males moved further than what is typically observed in females of the species, survival was considerably lower. These unique insights improve our understanding of mating interactions in wild Ae. aegypti populations and lay the foundation for robust suppression strategies in the future.

A new strategy for constructing artificial light-harvesting systems: supramolecular self-assembly gels with AIE properties

Soft Matter ◽  
2021 ◽  
Author(s):  
Xinxian Ma ◽  
jinlong le ◽  
Yipei Wang ◽  
Yang Gao ◽  
bo qiao ◽  
...  
Keyword(s):  
Rhodamine B ◽  
Self Assembly ◽  
Light Harvesting ◽  
Benzimidazole Derivative ◽  
Fluorescent Dye ◽  
Artificial Light ◽  
New Strategy ◽  
Harvesting Systems

An artificial light-harvesting system (ALHS) has been designed and constructed based on supramolecular organogels of a simple hydrazide-functionalized benzimidazole derivative (HB), as well as fluorescent dye Rhodamine B (RhB). RhB...

Temperature Measurement in Microfluidic Systems Using Photobleaching of a Fluorescent Slab

2008 ◽  
Author(s):  
Lin Gui ◽  
Carolyn L. Ren
Keyword(s):  
Temperature Distribution ◽  
Temperature Measurement ◽  
Rhodamine B ◽  
Fluorescent Dye ◽  
Measured Temperature ◽  
Pdms Layer ◽  
Temperature Dependent ◽  
Buffer Solution ◽  
Fluorescent Intensity ◽  
Dye Diffusion

Temperature control is key to microfluidic-based Lab-on-a-Chip devices for a variety of applications such as polymerase chain reaction for DNA amplification and isoelectric focusing for protein separation where pH gradients are thermally generated. The most widely used temperature measurement method involves the mixing of the buffer solution with a fluorescent dye, which has a temperature-dependent fluorescent intensity. The temperature distribution in the liquid can be obtained by monitoring the fluorescent intensity distribution in the channel. However, this method can not be easily applied to polymer-made microfluidic chips because of dye absorption and penetration into polymer chips, electrophoresis of dye which causes artificial temperature gradients, and inevitable photobleaching of fluorescent dye. Therefore, a novel method is developed and presented here for temperature measurement by utilizing photobleaching of fluorescent dye. This method includes two novel contributions: i) a specially developed model for converting temperature-dependent photobleaching speed distribution to temperature distribution, and ii) an introduction of a thin polydimethylsiloxane (PDMS) layer with saturated Rhodamine B for solving the above-mentioned dye diffusion and electrophoresis problems. In this new method, a thin PDMS layer saturated with Rhodamine B is bonded with another PDMS layer with microchannels instead of mixing the dye with the buffer solution. Therefore, the problems associated with dye diffusion into PDMS chips and electrophoresis when an electrical field is applied to channels are avoided. The developed theory is validated by comparing the experimentally measured temperature distribution with numerical predicted results. The theory and its validation will be presented and discussed.

scholarly journals Fluorescent markers rhodamine B and uranine for Anopheles gambiae adults and matings

2020 ◽  
Author(s):  
Erica i Aviles ◽  
Rachel D Rotenberry ◽  
C Matilda Collins ◽  
Ellen M Dotson ◽  
Mark Q Benedict
Keyword(s):  
Anopheles Gambiae ◽  
Rhodamine B ◽  
Seminal Fluid ◽  
Adult Longevity ◽  
Potential Contribution ◽  
Laboratory Studies ◽  
Unique Identifier ◽  
Mating Competitiveness ◽  
Fluorescent Markers ◽  
Auto Fluorescence

Abstract Background Marking mosquitoes is vital for mark-release-recapture and many laboratory studies, but their small size precludes the use of methods that are available for larger animals such as unique identifier tags and radio devices. Fluorescent dust is the most commonly used method to distinguish released individuals from the wild population. Numerous colors and combinations can be used, however, dust sometimes affects longevity and behavior so alternatives that do not have these effects would contribute substantially. Rhodamine B has previously been demonstrated to be useful for marking adult Aedes aegypti males when added to the sugar meal. Unlike dust, this also marked the seminal fluid making it possible to detect matings by marked males in the spermatheca of females. Here we evaluate marking in Anopheles gambiae s. s. with rhodamine B and uranine to estimate their potential contribution. Methods Two fluorescent markers, rhodamine B and uranine, were dissolved in sugar water and fed to adult Anopheles gambiae. Concentrations that are useful for marking individuals and seminal fluid were determined. The effects on adult longevity, the durability of the marking and detection of the marker in mated females was determined. We also evaluated effects on mating competitiveness. Results Rhodamine B marking in adults is detectable for at least three weeks, however uranine marking declines with time and at low doses can be confused with auto-fluorescence. Both can be used for marking seminal fluid which can be detected in females mated by marked males, but, again, at low concentrations uranine-marking is more easily confused with the natural fluorescence of seminal fluid. Neither dye affected mating competitiveness. Conclusions Both markers tested could be useful for field and laboratory studies. Their use has substantial potential to contribute to a greater understanding of the bio-ecology of this important malaria vector. Rhodamine B has the advantage that it appears to be permanent and is less easily confused with auto-fluorescence. The primary limitation of both methods is that sugar feeding is necessary for marking and adults must be held for at least 2 nights to ensure all individuals are marked whereas dusts provide immediate and thorough marking.

scholarly journals Mark-release-recapture of male Aedes aegypti (Diptera: Culicidae): use of rhodamine B to estimate movement, mating and population parameters in preparation for an incompatible male program

2020 ◽  
Author(s):  
Brendan J. Trewin ◽  
Daniel Pagendam ◽  
Brian J. Johnson ◽  
Chris Paton ◽  
Nigel Snoad ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Adult Male ◽  
Rhodamine B ◽  
Field Experiments ◽  
Large Field ◽  
Adult Males ◽  
Sterile Male ◽  
Effective Population ◽  
Male Population ◽  
Mark Release Recapture

AbstractRapid advances in biological and digital technologies are revolutionizing the population control of invasive disease vectors such as Aedes aegypti. Methods such as the sterile and incompatible insect techniques (SIT/IIT) rely on modified males to seek out and successfully mate with females, and in doing so outcompete the wild male population for mates. Currently, these interventions infer the success of mating interactions between male and female insects through area-wide population surveillance and observations of mating competitiveness are rare. Furthermore, little is known about male Ae. aegypti behaviours and biology in field settings. In preparation for a large, community scale IIT program, we undertook a series of mark-release-recapture experiments using rhodamine B to mark male Ae. aegypti sperm and measure mating interactions with females. We also developed the Spatial and Temporally Evolving Isotropic Kernel (STEIK) framework to assist researchers to estimate the movement of individuals through space and time. Results showed that ~40% of daily females captured were unmated, suggesting interventions will need to release males regularly to be effective at suppressing Ae. aegypti populations. Males moved rapidly through the landscape, particularly when released during the night. Although males moved further than what is typically observed in females of the species, survival was considerably lower. These unique insights will lead to a greater understanding of mating interactions in wild insect populations and lay the foundation for robust suppression strategies in the future.Author SummaryModern scientific techniques for controlling populations of the dengue vector, Aedes aegypti, utilize the mating biology of adult male mosquitoes to achieve suppression through a sterilization process. As the study of Ae. aegypti control has typically focused on adult female mosquitoes, knowledge on male movement, survival and mating interactions in the field is lacking. Here we undertook several mark-release-recapture experiments on adult male Ae. aegypti in Innisfail, Australia, and measured important biological parameters. For the first time in large field experiments, we employed rhodamine B as a marker that when fed to adult males, identified both marked males and the wild females they mated with. We observed males moving further through the landscape, but surviving for a shorter period, than previous measurements undertaken on females in a field setting. A high proportion (~40%) of unmated females suggests individuals are constantly available for mating. As such, sterile male strategies may need to release at regular intervals to achieve effective population suppression. The unique insights provided by this study will assist in designing future sterile male field interventions.

Investigation of Developmental Stage/Age, Gamma Irradiation Dose, and Temperature in Sterilization of Male Aedes aegypti (Diptera: Culicidae) in a Sterile Insect Technique Program

2021 ◽  
Author(s):  
Beni Ernawan ◽  
Tjandra Anggraeni ◽  
Sri Yusmalinar ◽  
Intan Ahmad
Keyword(s):  
Aedes Aegypti ◽  
Gamma Irradiation ◽  
Sterile Insect Technique ◽  
Irradiation Dose ◽  
Developmental Stages ◽  
Pupal Stage ◽  
Standard Protocol ◽  
Mating Competitiveness ◽  
Sterile Insect Technique Program ◽  
High Level

Abstract The sterilization process using gamma irradiation is a crucial component in a program using sterile insect technique (SIT) to control Aedes aegypti. Unfortunately, there is no efficient standard protocol for sterilizing mosquitoes that can produce a high level of sterility while maintaining mating ability and longevity. Therefore, we conducted a study of the critical factors necessary to develop such a standard protocol. In this study, male Ae. aegypti pupae, as well as adults aged 1 d and 3 d, were irradiated using a Gamma-cell 220 irradiator doses of 0, 20, 40, 60, 70, 80, and 100 Gray (Gy). In addition, male Ae. aegypti in the pupal and adult stage aged 1 d were irradiated at a dose of 70 Gy at various temperatures. Changes in emergence rates, longevity, sterility, and mating competitiveness were recorded for each combination of parameters. Results showed that an increase of irradiation dose leads to a rise of induced sterility at all developmental stages, while simultaneously reducing emergence rate, survival, and mating competitiveness. Higher temperatures resulted in increased levels of sterility, reduced longevity, and did not affect the ability to mate. This study found that an irradiation dose of 70 Gy at a temperature between 20.00 and 22.30°C administered in the pupal stage induced a high level of sterility (around 98%), while maintaining mating competitiveness and longevity.

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