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 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 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 ◽  
Anopheles Gambiae S.S ◽  
Unique Identifier ◽  
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 Fluorescent markers rhodamine B and uranine for Anopheles gambiae adults and matings

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Erica I. Aviles ◽  
Rachel D. Rotenberry ◽  
C. Mathilda Collins ◽  
Ellen. M. Dotson ◽  
Mark Q. Benedict
Keyword(s):  
Anopheles Gambiae ◽  
Rhodamine B ◽  
Fluorescent Markers

scholarly journals Male Mating Rate Is Constrained by Seminal Fluid Availability in Bedbugs, Cimex lectularius

PLoS ONE ◽  
2011 ◽  
Vol 6 (7) ◽  
pp. e22082 ◽  
Author(s):  
Klaus Reinhardt ◽  
Richard Naylor ◽  
Michael T. Siva-Jothy
Keyword(s):  
Seminal Fluid ◽  
Cimex Lectularius ◽  
Male Mating ◽  
Mating Rate

scholarly journals Male Mating Competitiveness of a Wolbachia-Introgressed Aedes polynesiensis Strain under Semi-Field Conditions

2011 ◽  
Vol 5 (8) ◽  
pp. e1271 ◽  
Author(s):  
Eric W. Chambers ◽  
Limb Hapairai ◽  
Bethany A. Peel ◽  
Hervé Bossin ◽  
Stephen L. Dobson
Keyword(s):  
Field Conditions ◽  
Male Mating ◽  
Mating Competitiveness ◽  
Aedes Polynesiensis

The influence of late-stage pupal irradiation and increased irradiated: un-irradiated male ratio on mating competitiveness of the malaria mosquito Anopheles arabiensis Patton

2008 ◽  
Vol 99 (3) ◽  
pp. 317-322 ◽  
Author(s):  
M.E.H. Helinski ◽  
B.G.J. Knols
Keyword(s):  
Genetic Control ◽  
Anopheles Arabiensis ◽  
Pupal Stage ◽  
Male Mating ◽  
Malaria Mosquito ◽  
Large Cage ◽  
Mating Competitiveness ◽  
Threefold Increase ◽  
Large Numbers ◽  
Control Programmes

AbstractCompetitiveness of released males in genetic control programmes is of critical importance. In this paper, we explored two scenarios to compensate for the loss of mating competitiveness after pupal stage irradiation in males of the malaria mosquito Anopheles arabiensis. First, competition experiments with a higher ratio of irradiated versus un-irradiated males were performed. Second, pupae were irradiated just prior to emergence and male mating competitiveness was determined.Males were irradiated in the pupal stage with a partially or fully-sterilizing dose of 70 or 120 Gy, respectively. Pupae were irradiated aged 20–26 h (young) as routinely performed, or the pupal stage was artificially prolonged by cooling and pupae were irradiated aged 42–48 h (old). Irradiated males competed at a ratio of 3:1:1 to un-irradiated males for mates in a large cage design.At the 3:1 ratio, the number of females inseminated by males irradiated with 70 Gy as young pupae was similar to the number inseminated by un-irradiated males for the majority of the replicates. At 120 Gy, significantly fewer females were inseminated by irradiated than by un-irradiated males. The irradiation of older pupae did not result in a significantly improved male mating competitiveness compared to the irradiation of young pupae.Our findings indicate that the loss of competitiveness after pupal stage irradiation can be compensated for by a threefold increase of irradiated males, but only for the partially-sterilizing dose. In addition, cooling might be a useful tool to facilitate handling processes of large numbers of mosquitoes in genetic control programmes.

scholarly journals Cross-species Y chromosome function between malaria vectors of the Anopheles gambiae species complex

2017 ◽  
Author(s):  
Federica Bernardini ◽  
Roberto Galizi ◽  
Mariana Wunderlich ◽  
Chrysanthi Taxiarchi ◽  
Nace Kranjc ◽  
...  
Keyword(s):  
Anopheles Gambiae ◽  
Y Chromosome ◽  
Hybrid Sterility ◽  
Substantial Reduction ◽  
Direct Consequence ◽  
Malaria Vectors ◽  
Evolutionary Divergence ◽  
Male Mating ◽  
Gambiae Complex ◽  
Male Hybrid

AbstractY chromosome function, structure and evolution is poorly understood in many species including the Anopheles genus of mosquitoes, an emerging model system for studying speciation that also represents the major vectors of malaria. While the Anopheline Y had previously been implicated in male mating behavior, recent data from the Anopheles gambiae complex suggests that, apart from the putative primary sex-determiner, no other genes are conserved on the Y. Studying the functional basis of the evolutionary divergence of the Y chromosome in the gambiae complex is complicated by complete F1 male hybrid sterility. Here we used an F1xF0 crossing scheme to overcome a severe bottleneck of male hybrid incompatibilities and enabled us to experimentally purify a genetically labelled A. gambiae Y chromosome in an A. arabiensis background. Whole genome sequencing confirmed that the A. gambiae Y retained its original sequence content in the A. arabiensis genomic background. In contrast to comparable experiments in Drosophila, we find that the presence of a heterospecific Y chromosome has no significant effect on the expression of A. arabiensis genes and transcriptional differences can be explained almost exclusively as a direct consequence of transcripts arising from sequence elements present on the A. gambiae Y chromosome itself. We find that Y hybrids show no obvious fertility defects and no substantial reduction in male competitiveness. Our results demonstrate that, despite their radically different structure, Y chromosomes of these two species of the gambiae complex that diverged an estimated 1.85Myr ago function interchangeably, thus indicating that the Y chromosome does not harbor loci contributing to hybrid incompatibility. Therefore, Y chromosome gene flow between members of the gambiae complex is possible even at their current level of divergence. Importantly, this also suggests that malaria control interventions based on sex-distorting Y drive would be transferable, whether intentionally or contingent, between the major malaria vector species.

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