Initial Post-Release Performance of Cultured Cyprinus chilia Juveniles in a Shallow Lake in Southwestern China.

The post-release performance of cultured fish is crucial for understanding the viability of cultured fish and assessing the effects of stock enhancement programs. This study aimed to investigate the initial post-release performance of cultured Cyprinus chilia juveniles by examining their movement, spatial distribution, gut fullness, and gut microbiota in nature. In July 2022, a total of 20,000 C. chilia juveniles, tagged with visible implant fluorescence (VIE), were released into Qilu Lake, a shallow lake in southwestern China. Subsequently, continuous recapture was conducted at fixed recapture sites using trap nets during the first 7 days, one month and three months after release. Out of the released fish, 512 were recaptured, resulting in a recapture rate of 2.56%. The recaptured fish had a 100% tag retention rate. The majority (98.05%) of the recaptured fish were found in the recapture sites located on the eastern or western lakeshore, while only 10 fish were recaptured from the recapture sites in the northern lake area. The water depth range where the recaptured fish were found ranged from 190 to 350 cm, with most fish preferring depths less than 300 cm. The majority of the released fish migrated towards the eastern and western lakeshore, with long-distance movement (greater than 100 m) primarily occurring within the first four days after release. The level of gut fullness in the released fish initially decreased and then increased over time following release. Regarding gut microbiota, the dominant phyla observed in most samples were Firmicutes, Proteobacteria, Cyanobacteria, and Fusobacteria. Furthermore, significant variations in the dominant genera were observed across different samples. Principal coordinates analysis (PCoA) revealed clear separation between the microbial communities of pre-release and post-release C. chilia juveniles. This study demonstrated that VIE tagging was a suitable method for short-term marking of C. chilia juveniles. Lakeshores with water depths less than 300 cm were identified as preferred habitats for C. chilia juveniles. The primary adaptation period for cultured C. chilia juveniles released into the natural environment was found to be approximately 4-5 days. These findings contribute to our understanding of the post-release performance of cultured fish and may provide guidance for the management and evaluation of relevant stock enhancement programs.

Population structure of Phanaeus vindex (Coleoptera: Scarabaeidae) in SE Michigan.

Until now, little is known about the population structure and mobility of temperate dung beetles including the rainbow scarab, Phanaeus vindex (MacLeay 1819), although this knowledge is essential for their conservation as pastures become increasingly rare and the landscape fragmented by monocultures and urbanization. Here, we estimated population size, longevity, and dispersal within and between pastures. For 3 yr, we life-trapped beetles every week on 2 adjacent farms in SE Michigan, determined their sex, male morph, and size, and marked their elytra with individual tattoo patterns before releasing them. We marked a total of 470 rainbow scarabs of which 14 were recaptured once and 2 were recaptured twice. The sex ratio was not significantly sex-biased but fluctuated between months with no apparent uniformity between years. While the minor to major male ratios were unbiased in 2019 and 2020, they were marginally minor-biased in 2021. The gross population estimates for the 2 farms were 458-491 and 217 rainbow scarabs, respectively. Beetles traveled distances of up to 178 m within farms. No beetles dispersed between farms. One large female was recaptured after 338 days documenting the first cold hardiness and long lifespan of a cold-temperate dung beetle species in the wild. The low population estimates on both farms indicate 2 vulnerable populations with no or extremely limited connectivity. Supplementary funding for the land stewardship of small-scale cattle farmers could stabilize populations of native dung beetles and maintain their ecosystem services.

Moderate mobility and high density in a small area: the population ecology of the marsh fritillary Euphydryas aurinia in Transylvania (Romania).

The Natura 2000 network is one of the most important tools for nature conservation in the EU. Therefore, knowledge of the ecology of the species listed on the Habitats Directive annexes is of particular relevance. One of these species is the butterfly Euphydryas aurinia. Much is already known about the ecology of this species at the European level, but information on Romania is scarce. Therefore, in a study area northwest of Cluj, the species was intensively studied in a mark-release-recapture study over its entire flight period. The investigated population harboured many individuals and had a high density. The dispersal behaviour of the individuals was somewhat more pronounced than in Mediterranean and partly also high alpine populations, but roughly comparable to other populations of the nominotypic subspecies. Overall, the ecological differences to Central and Western European populations were not very pronounced, and many of the conservation-relevant statements on this species in Central Europe should also be largely valid in Romania.

A sterile insect technique pilot trial on Captiva Island: defining mosquito population parameters for sterile male releases using mark-release-recapture.

BACKGROUND: The sterile insect technique (SIT), which involves area-wide inundative releases of sterile insects to suppress the reproduction of a target species, has proven to be an effective pest control method. The technique demands the continuous release of sterilized insects in quantities that ensure a high sterile male:wild male ratio for the suppression of the wild population over succeeding generations. METHODS: For these releases, it is important to determine several ecological and biological population parameters, including the longevity of the released males in the field, the dispersal of the released males and the wild pest population size. The Lee County Mosquito Control District initiated a study in a 47-ha portion of Captiva Island (Florida, USA), an island with a total area of 230 ha, to define biological SIT parameters for Aedes aegypti (L.), an invasive disease-vectoring mosquito known to be difficult to control due to a combination of daytime biting activity, use of cryptic breeding habitats that are difficult to target with conventional night-time ultra-low volume methods, and emerging resistance to commonly used insecticides. Another goal was to assess patterns of dispersal and survival for laboratory-reared sterile Ae. aegypti males released over time in the pilot site. These parameters will be used to evaluate the efficacy of a SIT suppression program for Ae. aegypti on Captiva Island. RESULTS: Over the course of seven mark-release-recapture studies using single- and multiple-point releases, 190,504 sterile marked males were released, for which the recapture rate was 1.5% over a mean period of 12 days. The mean distance traveled by sterile males of the local strain of Ae. aegypti that has colonized Captiva Island was 201.7 m from the release point, with an observed maximum traveled distance of 404.5 m. The released sterile mosquitoes had a probability of daily survival of 0.67 and an average life expectancy of ~ 2.46 days. CONCLUSIONS: These data together with the population size estimate and sterile:wild ratio provide a solid basis for planning the SIT operational phase which is aimed at mosquito population suppression.

Simultaneous external and internal marking of Triatoma sordida nymphs: trace element efficacy and microgeographic dispersal in a peridomestic Brazilian Cerrado rural household.

BACKGROUND: Chagas disease (American trypanosomiasis) is an important neglected tropical illness, which has the flagellate protozoan Trypanosoma cruzi as etiological agent and blood-feeding insects of the Triatominae subfamily as vectors. Despite its importance for disease epidemiology, field studies targeting microgeographic dispersal of triatomines in endemic areas are rare. The ability wingless nymphs have to move (crawl) within peridomestic settings is a key component regarding the design and development of rational control strategies. METHODS: We double-marked Triatoma sordida fourth-instar nymphs (N4) with a reliable fluorescent dye and a trace element. This new methodology allowed us to simultaneously evaluate (i) nymph dispersal and (ii) the effectiveness of copper (Cu), chromium (Cr), and cadmium (Cd) trace elements as potential new markers. In the mark-release-recapture (MRR) experiment, 390 T. sordida N4 were released in the peridomicile of a single rural household, 130 individuals at each of three release points, at distances of 2, 5, and 10 m from the chicken coop (CC) and 27, 32, and 35 m away from the horse corral (HC). All specimens were double marked (Cu/blue, Cr/orange, Cd/green). Recaptures occurred in two intervals: 1-3 days and 15-17 days after release. RESULTS: Specimens were successfully recaptured at all distances up to 10 m. A total of 19, 23, and 10 specimens were able to disperse 2, 5, and 10 m, respectively, to reach the CC. No insects were recaptured at the HC. Of the three analyte/paint combinations tested, Cr/orange gave the most promising results; Cu/blue marker and Cd/green marker performed very poorly with only 4/19 and 0/10 analyte/paint ratios, respectively. CONCLUSIONS: Triatoma sordida N4 could cover a distance of 10 m in 17 days. This indicates that nymphs seem to have a reduced dispersal capability compared to adults. Ninety-one percent of the 22 recaptured orange-marked nymphs were still Cr positive after the 17-day period evaluated. This makes this analyte a good candidate for future investigations that will apply this marking method in MRR studies.

Effects of SmartWater, a fluorescent mark, on the dispersal, behavior, and biocontrol efficacy of Phytoseiulus persimilis.

New marking methods for studying small biocontrol agents (especially predatory mites) are needed because many current techniques are expensive, ineffective or not applicable to small organisms. The objective of this study was to determine whether SmartWater, a liquid and permanent fluorescent dye, can be used to mark Phytoseiulus persimilis for experimentation without any deleterious effects on its dispersal, behavior, reproduction, and biocontrol efficacy. Our results show that there were no significant differences in movement, inter-plant dispersal, feeding behavior, survivability, and reproduction between marked P. persimilis and control individuals sprayed with water. We also found that the SmartWater mark lasted for the duration of the mites' life, indicating strong durability over time. Marking efficacy may be reduced, due to a trade-off between batch marking efficacy and the possibility of drowning study organisms. However, we feel future research could improve liquid marking techniques that would reduce this risk. Overall, this study concludes that SmartWater could be a useful marking tool for predatory mites in both laboratory and field studies.

Field Suppression of Spotted Wing Drosophila (SWD) (Drosophila suzukii Matsumura) Using the Sterile Insect Technique (SIT).

Drosophila suzukii (spotted wing drosophila­SWD) is an economically important pest of soft and stone fruit worldwide. Control relies on broad-spectrum insecticides, which are neither fully effective nor environmentally sustainable. The sterile insect technique (SIT) is a proven, effective and environmentally friendly pest-management tool. Here, we investigated, for the first time, the potential of using SIT to control D. suzukii in field conditions without physical barriers that limit insect invasion. A proprietary method of rearing and irradiation with X-rays was used to obtain males that were > 99% sterile. Sterile males were released twice per week from April to October 2021 on a site in Kent, UK, where everbearing strawberries were grown in open polytunnels. The infestation of wild female D. suzukii was monitored weekly using red sticky traps with dry lure at the treated site and at two similar control sites that did not receive sterile male releases. Releases of sterile males suppressed the wild female D. suzukii population by up to 91% in comparison with the control sites. We thus demonstrated the feasibility of SIT to achieve season-long control of D. suzukii using early, sustained and dynamically targeted releases of sterile males. This provides a promising environmentally friendly method to control this important pest.

Source or Sink? The Role of Residential Host Plants in Asian Citrus Psyllid Infestation of Commercial Citrus Groves.

Citriculture landscapes in the U.S. are typically habitat mosaics of commercial groves interspersed with residential areas supporting a variety of unmanaged citrus. Diaphorina citri the vector of Candidatus Liberibacter asiaticus, the causal agent of Huanglongbing feeds on citrus in both habitats. We postulated that residential citrus function as a 'source' of D. citri that infest groves, functioning as sinks. Here we report on an experimental mark-release-recapture study conducted at the interface of a residential neighborhood and groves. Adult D. citri marked with colored fluorescent powders were released in both habitats (n = 15,300) and their movement within and between milieus monitored. Although the recapture rate of marked psyllids was very low (0.23%), the results were instructive. Most of the recaptured psyllids in residential trees (84.6%) were released within that habitat. In contrast, approximately half of the marked psyllids recovered in groves were released in residential areas. Of all the recaptured psyllids, about 40% changed habitats, but the change was skewed toward movement from residential to grove habitat. These data strongly suggest that there is a constant exchange of D. citri adults between the two habitats, with residential citrus trees functioning as a source habitat of psyllids. The further the residential trees are located from groves, the less likely they will serve as sources of D. citri. Hence, to reduce the risks of citrus grove colonization by D. citri, new groves should be established away from residential habitats where possible, and psyllid management practices must also be implemented in residential habitats.

Biological Aspects and Movements of Neotropical Fruit-Feeding Butterflies.

The patterns of insect movement are the cumulate product of many individual decisions and can be shaped by the way morphology and behaviour interacts with landscape structure and composition. Hence, the ongoing process of habitat fragmentation increases the distance among suitable habitats and changes those in such a way that it may favour some movement behaviour. Our study described some biological traits (sex ratio, age structure and individual permanence in a population), as well as the movements of fruit-feeding butterflies along the horizontal dimension (among habitats: forest interior, natural forest transition - ecotone and anthropogenic forest transition - edge) and the vertical dimension (between canopy and understory). We sampled butterflies monthly over 1 year in the Atlantic rainforest, South-eastern Brazil, following a standardized design using bait traps. We found that sex ratio was male biased for five out of the six more abundant species and the age structure showed an increase in recruitment of new individuals in the dry season followed by a noticeable aging of the populations in the wet season. In general, our results revealed an aggregated spatial distribution, in which few individuals travelled long distances while most individuals were recaptured in the same trap, suggesting that all studied habitats currently provide the necessary conditions for the maintenance of butterfly populations, favouring fewer movements and narrow home ranges for both sexes and species. Conservation of a set of heterogeneous habitats it is especially important for the maintenance of sedentary butterflies and of those that move large distances.

Mosquito tagging using DNA-barcoded nanoporous protein microcrystals.

Conventional mosquito marking technology for mark-release-recapture (MRR) is quite limited in terms of information capacity and efficacy. To overcome both challenges, we have engineered, lab-tested, and field-evaluated a new class of marker particles, in which synthetic, short DNA oligonucleotides (DNA barcodes) are adsorbed and protected within tough, crosslinked porous protein microcrystals. Mosquitoes self-mark through ingestion of microcrystals in their larval habitat. Barcoded microcrystals persist trans-stadially through mosquito development if ingested by larvae, do not significantly affect adult mosquito survivorship, and individual barcoded mosquitoes are detectable in pools of up to at least 20 mosquitoes. We have also demonstrated crystal persistence following adult mosquito ingestion. Barcode sequences can be recovered by qPCR and next-generation sequencing (NGS) without detectable amplification of native mosquito DNA. These DNA-laden protein microcrystals have the potential to radically increase the amount of information obtained from future MRR studies compared to previous studies employing conventional mosquito marking materials.

Coping with Environmental Extremes: Population Ecology and Behavioural Adaptation of Erebia pronoe, an Alpine Butterfly Species.

A mark-recapture study of the nominotypical Erebia pronoe in the Alps was conducted to survey its ecological demands and characteristics. Population structure analysis revealed a combination of protandry (one-week earlier eclosion of males) and serial eclosion. Significant differences between both sexes were found in population density (males: 580/ha ± 37 SE; females: 241/ha ± 66 SE), sex-ratio (2.4) and behaviour (57.7 vs. 11.9% flying). Both sexes used a wide range of nectar plants (Asteraceae, 77.3%; Dipsacaceae, 12.3%; Gentianaceae, 9.7%). The use of nectar plants shows a non-specific spectrum, which, however, completely avoids overlap with the locally co-occurring species Erebia nivalis. Movement patterns show the establishment of homeranges, which significantly limits the migration potential. Due to its broad ecological niche, E. pronoe will probably be able to react plastically to the consequences of climate change. The formation of high population densities, the unconcerned endangerment status, the unspecific resource spectrum and the sedentary character of the species make E. pronoe a potential indicator of the quality and general resource occurrence of alpine rupicolous grasslands.

A novel fluorescence and DNA combination for versatile, long-term marking of mosquitoes.

Current mark-release-recapture methodologies are limited in their ability to address complex problems in vector biology, such as studying multiple groups overlapping in space and time. Additionally, limited mark retention, reduced post-marking survival and the large effort in marking, collection and recapture all complicate effective insect tracking.We have developed and evaluated a marking method using a fluorescent dye (SmartWater®) combined with synthetic DNA tags to informatively and efficiently mark adult mosquitoes using an airbrush pump and nebulizer. Using a handheld UV flashlight, the fluorescent marking enabled quick and simple initial detection of recaptures in a field-ready and non-destructive approach that when combined with an extraction-free PCR on individual mosquito legs provides potentially unlimited marking information.This marking, first tested in the laboratory with Anopheles gambiae s.l. mosquitoes, did not affect survival (median ages 24-28 days, p-adj > 0.25), oviposition (median eggs/female of 28.8, 32.5, 33.3 for water, green, red dyes, respectively, p-adj > 0.44) or Plasmodium competence (mean oocysts 5.56-10.6, p-adj > 0.95). DNA and fluorescence had 100% retention up to 3 weeks (longest time point tested) with high intensity, indicating marks would persist longer.We describe a novel, simple, no/low-impact and long-lasting marking method that allows separation of multiple insect subpopulations by combining unlimited length and sequence variation in the synthetic DNA tags. This method can be readily deployed in the field for marking multiple groups of mosquitoes or other insects.

Bulk-up synchronization of successive larval cohorts of Anopheles gambiae and Anopheles coluzzii through temperature reduction at early larval stages: effect on emergence rate, body size and mating success.

BACKGROUND: Malaria persists as a huge medical and economic burden. Although the number of cases and death rates have reduced in recent years, novel interventions are a necessity if such gains are to be maintained. Alternative methods to target mosquito vector populations that involve the release of large numbers genetically modified mosquitoes are in development. However, their successful introduction will require innovative strategies to bulk-up mosquito numbers and improve mass rearing protocols for Anopheles mosquitoes. METHODS: The relationship between mosquito aquatic stage development and temperature was exploited so that multiple cohorts of mosquitoes, from separate egg batches, could be synchronized to 'bulk-up' the number of mosquitoes released. First instar larvae were separated into two cohorts: the first, maintained under standard insectary conditions at 27oC, the second subjected to an initial 5-day cooling period at 19oC. RESULTS: Cooling of 1st instars slowed the mean emergence times of Anopheles coluzzii and Anopheles gambiae by 2.4 and 3.5 days, respectively, compared to their 27oC counterparts. Pupation and emergence rates were good (> 85 %) in all conditions. Temperature adjustment had no effect on mosquito sex ratio and adult fitness parameters such as body size and mating success. CONCLUSIONS: Bulk-up larval synchronization is a simple method allowing more operational flexibility in mosquito production towards mark-release-recapture studies and mass release interventions.

Dispersal of Philaenus spumarius (Hemiptera: Aphrophoridae), a Vector of Xylella fastidiosa, in Olive Grove and Meadow Agroecosystems.

The introduction of the Xylella fastidiosa Wells bacterium into Apulia (South Italy) has caused the massive dieback of olive trees, and is threatening olive production throughout the Mediterranean Region. The key vector of X. fastidiosa in Europe is the spittlebug Philaenus spumarius L. The dispersal capabilities of P. spumarius are poorly known, despite being a key parameter for the prediction of the spread of the bacterium. In this study, we have examined the dispersal of P. spumarius adults in two different agroecosystems in Italy: an olive grove in Apulia (Southern Italy) and a meadow in Piedmont (Northern Italy). Insects were marked with albumin and released during seven independent trials over 2 yr. The recapture data were pooled separately for each agroecosystem and used to estimate the dispersal kernels of P. spumarius in the olive grove and in the meadow. The diffusion coefficient estimate for P. spumarius was higher in the meadow than in the olive grove. The median distance from the release point for 1 d of dispersal was 26 m in the olive grove and 35 m in the meadow. On the basis of our model, we estimated that 50% of the spittlebug population remained within 200 m (98% within 400 m) during the 2 mo period of high abundance of the vector on olives in Apulia. The dispersal of P. spumarius is thus limited to some hundreds of meters throughout the whole year, although it can be influenced to a great extent by the structure of the agroecosystem.

The Influence of Marking Methods on Mobility, Survivorship, and Field Recovery of Halyomorpha halys (Hemiptera: Pentatomidae) Adults and Nymphs.

Halyomorpha halys (Stål), the brown marmorated stink bug, is an invasive and highly polyphagous insect that has caused serious economic injury to specialty and row crops in the United States and Europe. Here, we evaluated the effects of marking adult and nymphal H. halys with four different colors of fluorescent powder (Blaze Orange, Corona Pink, Horizon Blue, and Signal Green) on mobility and survivorship in laboratory bioassays. Adults and nymphs were marked using liquified fluorescent powder solutions and allowed to dry prior to bioassay. The presence of the marking solution had no significant effects on adult or nymphal mobility, adult survivorship, nymphal development, or adult flight capacity. We also evaluated the persistence of neon marker applied to the pronotum of H. halys adults and found this technique remained detectable for 2 wk under field conditions. Although both marking techniques are inexpensive, persist for ≥1 wk, and do not affect mortality, the neon marker method is more time-consuming, taking ~12 times longer to mark 50 adult H. halys compared with the liquified fluorescent powders. Thus, we would recommend using fluorescent powders for large-scale mark-release-recapture studies.

Interpreting Temporal and Spatial Variation in Spotted-Wing Drosophila (Diptera: Drosophilidae) Trap Captures in Highbush Blueberries.

Integrated pest management (IPM) programs for the spotted-wing drosophila Drosophila suzukii (Diptera: Drosophilidae) rely on insecticide applications to reduce adult populations and prevent fruit infestation. Although monitoring traps are used for early D. suzukii adult detection to time the start of insecticide applications, it remains unclear whether trap counts can be used to determine the efficacy of these programs and predict the risk of fruit infestation. To address this, a 2-yr study (2016-2017) was conducted in highbush blueberries in New Jersey (USA) to interpret D. suzukii trap count variation in relation to the frequency of insecticide applications and proximity to forest habitats. We also correlated trap counts with fruit infestation and used traps to determine the maximum dispersive distance traveled by D. suzukii adults within blueberry fields by using mark-release-capture studies. Using a trapping network across nine farms, we demonstrated that insecticide applications reduce D. suzukii trap counts, but this varied according to seasonality, and that traps placed closer to forest habitats within farms had higher fly counts than those placed in farm interiors. Moreover, blueberry fields that had zero fruit infestation also had predictably lower trap counts than fields with infested fruit, and the maximum dispersive distance for D. suzukii within blueberry fields was 90 m. In summary, while D. suzukii trap counts in blueberry farms could predict the frequency of insecticide applications and fruit infestation, the predictive power of our trap data was too variable across the blueberry harvest period to make it a reliable tool.

A standardised method of marking male mosquitoes with fluorescent dust.

BACKGROUND: Prior to a major release campaign of sterile insects, including the sterile insect technique, male mosquitoes must be marked and released (small scale) to determine key parameters including wild population abundance, dispersal and survival. Marking insects has been routinely carried out for over 100 years; however, there is no gold standard regarding the marking of specific disease-transmitting mosquitoes including Anopheles arabiensis, Aedes aegypti and Aedes albopictus. The research presented offers a novel dusting technique and optimal dust colour and quantities, suitable for small-scale releases, such as mark-release-recapture studies. METHODS: We sought to establish a suitable dust colour and quantity for batches of 100 male An. arabiensis, that was visible both by eye and under UV light, long-lasting and did not negatively impact longevity. A set of lower dust weights were selected to conduct longevity experiments with both Ae. aegypti and Ae. albopictus to underpin the optimal dust weight. A further study assessed the potential of marked male An. arabiensis to transfer their mark to undusted males and females. RESULTS: The longevity of male An. arabiensis marked with various dust colours was not significantly reduced when compared to unmarked controls. Furthermore, the chosen dust quantity (5 mg) did not negatively impact longevity (P = 0.717) and provided a long-lasting mark. Dust transfer was found to occur from marked An. arabiensis males to unmarked males and females when left in close proximity. However, this was only noticeable when examining individuals under a stereomicroscope and thus deemed negligible. Overall, male Ae. aegypti and Ae. albopictus displayed a greater sensitivity to dusting. Only the lowest dust weight (0.5 mg) did not significantly reduce longevity (P = 0.888) in Ae. aegypti, whilst the lowest two dust weights (0.5 and 0.75 mg) had no significant impact on longevity (P = 0.951 and 0.166, respectively) in Ae. albopictus. CONCLUSION: We have devised a fast, inexpensive and simple marking method and provided recommended dust quantities for several major species of disease-causing mosquitoes. The novel technique provides an evenly distributed, long-lasting mark which is non-detrimental. Our results will be useful for future MRR studies, prior to a major release campaign.

Marking Drosophila suzukii (Diptera: Drosophilidae) with Fluorescent Dusts.

The marking of Drosophila suzukii can be an important instrument for studying the ecology and behaviour of this economically important fruit pest, aiding the development of new Integrated Pest Management (IPM) tools or strategies. There is, however, a need for a cost-effective methodology that provides an easily detectable and stable mark. Whereas fluorescent pigment powders are often used in entomological research, the pigments (series, dyes), application techniques, or doses need to be evaluated for each studied species in terms of their efficacy and possible adverse effects on the performance of the insect. The effectiveness of different application techniques and dyes (RadGlo® TP-series) and their effect on the survival of adult D. suzukii were investigated in the laboratory. Furthermore, the influence of the marking on the behaviour of the flies was examined in laboratory trap assays (olfaction) and a field recapture study (general orientation). The persistence and detectability of the marks was evaluated both on living flies (for different application techniques) and dead flies under trapping/storage conditions. The use of fluorescent powders to mark D. suzukii flies yielded a clearly detectable and highly persistent mark, without any adverse effects on the survival and behaviour of the flies.

Recaptures of feral Bactrocera dorsalis and B. umbrosa (Diptera: Tephritidae) males after feeding on methyl eugenol.

Two major fruit fly pest species, Bactrocera dorsalis and B. umbrosa, are strongly attracted to methyl eugenol (ME) found in >450 plant species. They are, however, exclusive pollinators of certain daciniphilous (attracting Dacini fruit flies) Bulbophyllum orchids. A comparison between the recaptures of feral males after feeding ad libitum on 0.6 mg ME (simulating an average floral quantity of an orchid flower - Trial 1) and 480 mg in Trial 2 was investigated using the non-invasive capture-mark-release-recapture (CMRR) technique. Based on daily CMRR over a 16-day period, using a different colour enamel paint each day, percentages of B. dorsalis males recaptured in Trial 1 were significantly higher than those in Trial 2. However, for B. umbrosa, percentages of recaptures for different day-specific colours were highly variable due to low fly numbers captured/day. In Trial 1, of 756 B. dorsalis males released, 36.4% were recaptured once, 7.7 twice, 2.4 three times and 0.4 four times. While in Trial 2 of 1157 released males, 6% were recaptured once and 0.3% twice. Of 67 B. umbrosa males released, 28.4% were recaptured once and none more than once in Trial 1. Nevertheless, of 119 flies released in Trial 2, 25.2% were recaptured once and 3.3% twice. Overall, many marked males did return to a single ME-source to 'refuel' ME (a sex pheromone precursor). The results also show that a relatively high number of flies paid multi-visitations to a single 0.6 mg ME-source and indicate that the presence of natural ME-sources may impact area-wide IPM programmes.

The development and evaluation of a self-marking unit to estimate malaria vector survival and dispersal distance.

BACKGROUND: A clear understanding of mosquito biology is fundamental to the control efforts of mosquito-borne diseases such as malaria. Mosquito mark-release-recapture (MMRR) experiments are a popular method of measuring the survival and dispersal of disease vectors; however, examples with African malaria vectors are limited. Ethical and technical difficulties involved in carrying out MMRR studies may have held back research in this area and, therefore, a device that marks mosquitoes as they emerge from breeding sites was developed and evaluated to overcome the problems of MMRR. METHODS: A modified self-marking unit that marks mosquitoes with fluorescent pigment as they emerge from their breeding site was developed based on a previous design for Culex mosquitoes. The self-marking unit was first evaluated under semi-field conditions with laboratory-reared Anopheles arabiensis to determine the marking success and impact on mosquito survival. Subsequently, a field evaluation of MMRR was conducted in Yombo village, Tanzania, to examine the feasibility of the system. RESULTS: During the semi-field evaluation the self-marking units successfully marked 86% of emerging mosquitoes and there was no effect of fluorescent marker on mosquito survival. The unit successfully marked wild male and female Anopheles gambiae sensu lato (s.l.) in sufficiently large numbers to justify its use in MMRR studies. The estimated daily survival probability of An. gambiae s.l. was 0.87 (95% CI 0.69-1.10) and mean dispersal distance was 579 m (95% CI 521-636 m). CONCLUSIONS: This study demonstrates the successful use of a self-marking device in an MMRR study with African malaria vectors. This method may be useful in investigating population structure and dispersal of mosquitoes for deployment and evaluation of future vector control tools, such as gene drive, and to better parameterize mathematical models.