Host plant ranges of fruit flies (Diptera: Tephritidae) in Madagascar.

Agriculture is one of the major farming activities, representing 32% of the gross domestic product of Madagascar and 74.3% of the population is involved in this activity. Fruit flies of the Tephritidae family are considered as the most destructive pests for agriculture in the country, nevertheless, few data exist on host plants and distribution of those pests. In the present study, we address those questions by conducting a large survey between November 2016 and July 2018 across the six agroecological regions of Madagascar. Fruit and vegetable were sampled from 198 plant species (wild and cultivated) and represented 37,965 fruits from all regions of Madagascar. The infestation index ranged from 0.06 to 538.46 pupae/kg, the infestation percentage was up to 54.84% in some samples, 63 plant species were considered as host of Tephritidae. Twelve fruit fly species were identified, seven of which were previously described as endemic, five species could be considered as widespread (altitudinal gradients between 1 and 1634 m asl) and major pests in Madagascar: Ceratitis malgassa (23 plant species from 12 families), Neoceratitis cyanescens (16 plant species from one family), Bactrocera dorsalis (18 plant species from 12 families), Dacus demmerezi (ten plant species from one family), Dacus vertebratus (six species from one family). Those results are of importance for implementation of control measures.

Genetic structure and range expansion of Zeugodacus Cucurbitae (Diptera: Tephritidae) in Africa.

Hypotheses about the worldwide colonization routes of the melon fly, Zeugodacus cucurbitae (Diptera: Tephritidae), are mainly based on sparse historical records. Here we aim at reconstructing the colonization history of the African continent based on an improved description of the population structure of Z. cucurbitae and approximate Bayesian analyses. Individuals of Z. cucurbitae were sampled in 17 localities from East, West and Central Africa and genotyped at 19 microsatellite markers. Bayesian analyses showed intracontinental population structuring with populations from Uganda diverging from those of Tanzania and populations from Burundi and Kenya showing traces of admixture with West African samples. Approximate Bayesian Computation provided support to the hypothesis of a single introduction Z. cucurbitae into East Africa and subsequent expansion to West Africa, each colonization event was followed by a bottleneck that promoted population divergence within Africa. Parameter estimates suggested that these events are roughly compatible with the historical records of Z. cucurbitae presence in sub-Saharan Africa (viz. 1936 in East Africa and 1999 in West Africa) and allow excluding alternative hypotheses on older or multiple introductions of Z. cucurbitae.

An integrated diagnostic setup for the morphological and molecular identification of the Ceratitis FAR complex (C. anonae, C. fasciventris, C. rosa, C. quilicii, Diptera, Tephritidae).

The Ceratitis FAR complex (Diptera, Tephritidae) includes four economically important frugivorous flies (Ceratitis anonae, Ceratitis fasciventris, Ceratitis quilicii, Ceratitis rosa) whose immature stages and adult females cannot be properly resolved through morphological identification. In order to develop a simplified molecular tool for the identification of two of these species (C. rosa, C. quilicii), we selected a subset of six microsatellite markers out of a panel of 16 loci that were previously developed for the molecular differentiation of the taxa within the complex. These six markers were first tested in silico and then used for the actual genotyping of C. quilicii and C. rosa, resulting in the correct identification of all male reference specimens. Here, we propose an integrated morphological and molecular setup for the identification of the four species of the FAR complex. The decision map relies on preliminary DNA barcoding or morphological identification (when possible) to exclude species not belonging to the complex followed by (a) morphological identification of all adult male specimens and female C. anonae, (b) molecular identification via a panel of 16 microsatellite markers for immature stages, damaged vouchers and samples potentially including adult female C. fasciventris/C. quilicii/C. rosa and (c) molecular identification via a reduced panel of six microsatellite markers for samples including only C. quilicii and C. rosa. This simplified diagnostic setup was profitably implemented in the framework of the ERAfrica fruit fly project and will help correctly identify species within the FAR complex for their early detection and monitoring.

Cassava whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in East African farming landscapes: a review of the factors determining abundance.

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a pest species complex that causes widespread damage to cassava, a staple food crop for millions of households in East Africa. Species in the complex cause direct feeding damage to cassava and are the vectors of multiple plant viruses. Whilst significant work has gone into developing virus-resistant cassava cultivars, there has been little research effort aimed at understanding the ecology of these insect vectors. Here we assess critically the knowledge base relating to factors that may lead to high population densities of sub-Saharan African (SSA) B. tabaci species in cassava production landscapes of East Africa. We focus first on empirical studies that have examined biotic or abiotic factors that may lead to high populations. We then identify knowledge gaps that need to be filled to deliver sustainable management solutions. We found that whilst many hypotheses have been put forward to explain the increases in abundance witnessed since the early 1990s, there are little published data and these tend to have been collected in a piecemeal manner. The most critical knowledge gaps identified were: (i) understanding how cassava cultivars and alternative host plants impact population dynamics and natural enemies; (ii) the impact of natural enemies in terms of reducing the frequency of outbreaks and (iii) the use and management of insecticides to delay the development of resistance. In addition, there are several fundamental methodologies that need to be developed and deployed in East Africa to address some of the more challenging knowledge gaps.

Invasion by Bactrocera dorsalis and niche partitioning among tephritid species in Comoros.

Ten economically important species belonging to the Tephritidae have been recorded in Union of the Comoros (an island nation off the coast of East Africa). Little is known about the distribution of these species and how they are affected by climatic factors in the Comoros archipelago. The main objectives of this study were to characterize: (i) the population dynamics of tephritid fruit flies in relation to season and host fruit availability and (ii) the geographic distribution of tephritids in relation to temperature and rainfall. The study was conducted during 2 years at 11 sites on three islands (Grande Comore, Anjouan, and Mohéli) in the archipelago. The site elevations ranged from 55 to 885 m a.s.l. At each site, flies were collected weekly in eight traps (four different lures, each replicated twice). Fruit phenology was also recorded weekly. The dominant tephritid species detected was the invasive Bactrocera dorsalis Hendel followed by Ceratitis capitata Wiedemann. Tephritid species were generally more abundant during the hot and rainy seasons than during the cold and dry seasons. Bactrocera dorsalis numbers were higher on Grande Comore than on the two other islands. On Anjouan and Mohéli, B. dorsalis numbers were very low in 2014 but sharply increased in 2015, suggesting a recent invasion of these islands. Abundances were significantly related to the fruiting of mango, strawberry guava, and guava for B. dorsalis and to the fruiting of mango, guava, and mandarin for C. capitata. Bactrocera dorsalis was more abundant in hot and humid low-altitude areas, while C. capitata was more abundant in dry medium-altitude areas, suggesting the occurrence of climatic niche partitioning between the two species.

Isolation and Characterization of 11 Polymorphic Microsatellite Markers Developed for Orthops palus (Heteroptera: Miridae).

Miridae (Hemiptera: Heteroptera: Cimicomorpha), or plant bugs, are one of the most diverse and species-rich families of insects. Most of them are phytophagous, but some are insect predators and used for biocontrol. Among this family, the mango bug, Orthops palus (Taylor 1947), is one of the most important pest of mango in Reunion Island. We developed 11 polymorphic microsatellite loci to study the population genetics of this pest species. The microsatellite markers were characterized by genotyping 78 field-collected insects sampled at different localities in Reunion Island. The number of alleles per locus ranged from 1 to 13 and heterozygosity levels ranged between 0.40 and 0.94. Several loci were not at Hardy-Weinberg equilibrium for the tested populations. These markers are the first to be developed for a species of the genus Orthops.

Assessing hygienic behavior of Apis mellifera unicolor (Hymenoptera: Apidae), the endemic honey bee from Madagascar.

Hygienic behavior (HB) is one of the natural mechanisms of honey bee for limiting the spread of brood diseases and Varroa destructor parasitic mite. Objective of our study was to measure HB of Apis mellifera unicolor colonies (N = 403) from three geographic regions (one infested and two free of V. destructor) in Madagascar. The pin-killing method was used for evaluation of the HB. Responses were measured from 3 h 30 min to 7 h after perforation of the cells. Colonies were very effective in detecting perforated cells. In the first 4 h, on average, they detected at least 50% of the pin-killed brood. Six hours after cell perforation, colonies tested (N = 91) showed a wide range of uncapped (0 to 100%) and cleaned cells (0 to 82%). Global distribution of the rate of cleaned cells at 6 h was multimodal and hygienic responses could be split in three classes. Colonies from the three regions showed a significant difference in HB responses. Three hypotheses (geographic, genetic traits, presence of V. destructor) are further discussed to explain variability of HB responses among the regions. Levels of HB efficiency of A. mellifera unicolor colonies are among the greatest levels reported for A. mellifera subspecies. Presence of highly hygienic colonies is a great opportunity for future breeding program in selection for HB.

Diapause incidence and duration in the pest mango blossom gall midge, Procontarinia mangiferae (Felt), on Reunion Island.

The mango blossom gall midge, Procontarinia mangiferae, is a multivoltine species that induces galls in inflorescences and leaves of the mango tree, Mangifera indica. In subtropical Reunion Island, populations of P. mangiferae are observed all-year round, but the pattern and the role of dormancy in their life cycle have never been documented. We performed field and laboratory experiments using more than 15,000 larvae. We demonstrated that a larval diapause may affect a part of the midge population, regardless of the season. The total duration of the diapause varied from 6 weeks to more than 1 year. One year of field monitoring showed that the highest incidence of diapause was observed in larvae collected during the summer from mango leaves, where it affected approximately one-third of the individuals. This facultative diapause allows the permanent presence of P. mangiferae in the orchards. By recording diapause duration during 22 weeks under controlled conditions, we showed that high temperatures (26 °C) increased diapause duration and extended the range of the dates of diapause emergence, whereas cool temperatures (20 °C) shortened diapause duration and shortened the range of the dates of emergence from diapause. A temperature decrease from 26 to 20 °C triggered the emergence of diapausing individuals. These mechanisms ensure the synchronization of the emergence of diapausing individuals with the appearance of mango inflorescences, which is also induced by cool winter temperatures.

Cryptic diversity and gene flow among three African agricultural pests: Ceratitis rosa, Ceratitis fasciventris and Ceratitis anonae (Diptera, Tephritidae).

The 'Ceratitis FAR complex' is a species complex of African fruit flies (Diptera, Tephritidae) including the major agricultural pest Ceratitis rosa and the morphologically similar Ceratitis fasciventris and Ceratitis anonae. To resolve their intra- and interspecific genetic relationships and to estimate gene flow within this complex, we surveyed allelic variation at 16 microsatellite loci in 27 African populations of the three morphospecies. Interpopulation genetic distances and individual Bayesian assignments distinguished five genotypic clusters: two involving C. rosa (R1, R2; that may occur in sympatry), two involving C. fasciventris (F1, F2; with parapatric distributions) and one involving C. anonae (A). Intra- and interspecific patterns of genetic differentiation were not hierarchically structured and genetic differentiation between conspecific clusters (F1-F2 and R1-R2) was higher or comparable with differentiation between heterospecific clusters (e.g. F1-A or R2-A). In some cases, gene flow estimates among morphospecies or among heterospecific genotypic clusters were significantly different from zero, showing the lack of reproductive isolation. Genetic differentiation between genotypic clusters was partly supported by morphological differences observed a posteriori in male secondary sexual characters. These results suggest important revisions to current models of ecological niche requirements and invasion risk of the major agricultural pest C. rosa and provide a basis for a taxonomic re-interpretation of the FAR complex.

Ecological interactions in Aedes species on Reunion Island.

Two invasive, container-breeding mosquito species, Aedes aegypti (Stegomyia aegypti) and Aedes albopictus (Stegomyia albopicta) (Diptera: Culicidae), have different distribution patterns on Reunion Island. Aedes albopictus occurs in all areas and Ae. aegypti colonizes only some restricted areas already occupied by Ae. albopictus. This study investigates the abiotic and biotic ecological mechanisms that determine the distribution of Aedes species on Reunion Island. Life history traits (duration of immature stages, survivorship, fecundity, estimated finite rate of increase) in Ae. aegypti and Ae. albopictus were compared at different temperatures. These fitness measures were characterized in both species in response to competitive interactions among larvae. Aedes aegypti was drastically affected by temperature, performing well only at around 25 °C, at which it achieved its highest survivorship and greatest estimated rate of increase. The narrow distribution of this species in the field on Reunion Island may thus relate to its poor ability to cope with unfavourable temperatures. Aedes aegypti was also more negatively affected by high population densities and to some extent by interactions with Ae. albopictus, particularly in the context of limited food supplies. Aedes albopictus exhibited better population performance across a range of environmental conditions. Its ecological plasticity and its superior competitive ability relative to its congener may further enhance its invasion success on Reunion Island.

Differential disease phenotype of begomoviruses associated with tobacco leaf curl disease in Comoros.

In the 2000s, tobacco plantations on the Comoros Islands were afflicted with a previously unobserved tobacco leaf curl disease characterised by symptoms of severe leaf curling and deformation. Previous molecular characterization of potential viral pathogens revealed a complex of African monopartite tobacco leaf curl begomovirus (TbLCVs). Our molecular investigation allowed the characterization of a new monopartite virus involved in the disease: tomato leaf curl Namakely virus (ToLCNamV). Agroinoculation experiments indicated that TbLCVs and tomato leaf curl viruses (ToLCVs) can infect both tomato and tobacco but that infectivity and symptom expression fluctuate depending on the virus and the plant cultivar combination.

Symbiont diversity and non-random hybridization among indigenous (Ms) and invasive (B) biotypes of Bemisia tabaci.

The whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is a worldwide pest and a vector of numerous plant viruses. B. tabaci is composed of dozens of morphologically indistinguishable biotypes and its taxonomic status is still controversial. This phloem-feeder harbours the primary symbiont Portiera aleyrodidarum and potentially six secondary symbionts: Cardinium, Arsenophonus, Hamiltonella, Rickettsia, Wolbachia and Fritschea. In the southwest Indian Ocean, La Réunion hosts two biotypes of this species: B (invasive) and Ms (indigenous). A multiplex PCR was developed to study the symbiont community of B. tabaci on La Réunion. Symbiont community prevalence and composition, host mitochondrial and nuclear genetic diversity, as well as host plant and localization, were described on field populations of La Réunion for B and Ms B. tabaci biotypes and their hybrids. A clear association between symbiotypes and biotypes was shown. Cardinium, Arsenophonus and Rickettsia were found in the Ms biotype (73.6%, 64.2% and 3.3%, respectively). Hamiltonella (exclusively) and Rickettsia were found in the B biotype (78% and 91.2%, respectively). Hybrids harboured all symbiotypes found in Ms and B populations, but with a higher prevalence of Ms symbiotypes than expected under random hybridization. An unexpected majority was Cardinium mono-infected (65.6%), and a striking minority (9%) harboured Cardinium/Arsenophonus. In the hybrids only, genetic diversity was linked to symbiotype. Among the hybrids, significant links were found between symbiotypes and: (i) mitochondrial COI sequences, i.e. maternal origin; and (ii) alleles of nuclear microsatellite loci, specific to either Ms or B parental biotype. Taken together, our results suggest that Cardinium and/or Arsenophonus may manipulate the reproduction of indigenous (Ms) with invasive (B) biotypes of Bemisia tabaci.

Genetic diversity, geographical range and origin of Bemisia tabaci (Hemiptera: Aleyrodidae) Indian Ocean Ms.

The whitefly Bemisia tabaci is a pest vector of begomoviruses on crops worldwide. Bemisia tabaci is composed of a complex of cryptic species which barely interbreed. An exception is the Ms from the South West Indian Ocean (SWIO), which crosses in low proportions with the exotic B. The Ms, together with B and Q is part of the same phylogenetic clad. To infer the genetic structure, the geographical range and putative origin of this putative species, microsatellite data and mitochondrial DNA (cytochrome oxydase I) sequences were analysed on an extensive sample set, including all the islands of the region and samples from mainland Africa. Only B and Ms populations were detected across these islands. The exotic B was found only on the islands of Réunion and Mauritius, whereas the Ms is found on all the SWIO islands. Very high isolation by distance was found for the Ms populations between islands of the SWIO, suggesting a long period of presence in this region. Ms populations from mainland Africa had a higher COI diversity than the Ms of the SWIO islands. This diversity is correlated with size and geological ages of the SWIO islands. The population genetic data obtained are in accordance with an origin of Ms in Africa, followed by its expansion and evolution across the SWIO islands prior to human arrival, confirming the status of Ms as indigenous in the SWIO islands.

Macrogeographic population structuring in the cosmopolitan agricultural pest Bactrocera cucurbitae (Diptera: Tephritidae).

The macrogeographic population structure of the agricultural pest Bactrocera cucurbitae (Diptera: Tephritidae) was investigated in order to identify the geographic origin of the species and reconstruct its range expansion. Individuals of B. cucurbitae were collected from 25 worldwide-distributed localities (n = 570) and genotyped at 13 microsatellite loci. The Bayesian clustering reveals that B. cucurbitae can be subdivided into five main groups corresponding to populations from (i) the African continent, (ii) La Réunion, (iii) Central Asia, (iv) East Asia and (v) Hawaii. The proportions of inter-regional assignments and the higher values of genetic diversity in populations from Pakistan, India and Bangladesh suggest that B. cucurbitae originated in Central Asia and expanded its range to East Asia and Hawaii on one hand and to Africa and the islands of the Indian Ocean on the other. A number of outliers (10-19 specimens according to different clustering algorithms) show high levels of admixture (Q > 0.70) with populations from different regions and reveal complex patterns of inter-regional gene flow. Anthropogenic transport is the most plausible promoter of this large-scale dispersal. The introduction of individuals from geographically distant sources did not have a relevant role in the most recent African invasions, which originated from the expansion of local populations. These results could provide a useful background to better evaluate invasion risks and establish priorities for the management of this cosmopolitan agricultural pest.

Influence of temperature on immature development, survival, longevity, fecundity, and gonotrophic cycles of Aedes albopictus, vector of chikungunya and dengue in the Indian Ocean.

Aedes albopictus is a mosquito originating from Asia, which has extended its range worldwide the last decades. It is a competent vector for several arboviruses. It was first described in La Réunion (an island of the South West part of the Indian Ocean) in 1913. Since then, it has become the dominant Aedes species and a serious threat to public health, especially during the two last arboviruses outbreaks of dengue (1977) and chikungunya (2005-2006). Despite its pest status, data on the biology of this vector are scarce, especially the population present in the Indian Ocean (IO), which has never been studied in detail. Therefore, the immature development, survival, longevity, fecundity, and gonotrophic cycles of Ae. albopictus were studied for an F2 population of the IO. These biological parameters were studied in controlled conditions at eight constant temperatures (5, 10, 15, 20, 25, 30, 35, and 40 degrees C). The minimal threshold of immature stages development was found at 10.4 degrees C and its optimum at 29.7 degrees C. The shortest periods for immature development were found at 30 degrees C, with in average of 8.8 d. The optimum intrinsic rate of growth (r) was observed between 25 and 30 degrees C. The gonotrophic cycles were also evaluated, and the shortest cycles were found at 30 degrees C (mean, 3.5 d). Those results are according to the field repartition of this species in La Réunion, allowing Ae. albopictus survival at a large range of temperatures.

Dispersal and survival of male and female Aedes albopictus (Diptera: Culicidae) on Réunion Island.

Mouse-baited traps were used to assess the longevity and dispersal of male and female Aedes albopictus by mark-release-recapture at two sites on La Réunion Island. Recapture rate was high, and mosquitoes of both sexes appeared up to 23 d after release. A daily survival probability of approximately 0.95 for males and females, far higher than expected, was estimated from these results. There was evidence that both sexes prefer to follow corridors of vegetation rather than crossing open spaces. Populations of wild mosquitoes had parous and insemination rates indicative of a young population. These results are relevant to future attempts to control this species by sterile insect technology.

Adaptation of the BG-Sentinel trap to capture male and female Aedes albopictus mosquitoes.

In recent years, the remarkable spread of Aedes albopictus (Skuse) (Diptera: Culicidae) throughout the world has drawn attention to this hitherto poorly studied species, particularly after its role in outbreaks of chikungunya fever in the western Indian Ocean and in Italy. Variants of sterile insect technique (SIT), including the release of transgenic males with a dominant lethal gene (RIDL), have been proposed in the search for new and innovative methods of control. Knowledge of male dispersal, mating behaviour and longevity will be critical to the success of this approach. We present an effective and practical method for trapping both male and female Ae. albopictus using a mouse-baited BG-Sentinel trap.

[First record of Aedes albopictus, vector of arboviruses in the Eparse Islands of the Mozambique Channel and updating of the inventory of Culicidae]. / Première mention d'Aedes albopictus, vecteur d'arbovirus, dans les îles Eparses du canal du Mozambique et actualisation de l'inventaire de la faune culicidienne.

This is the first description of Ae. albopictus in Grande Glorieuse and Juan de Nova, which are two isolated French islands of the Mozambique Channel. The recent introduction of this species is underlined by its absence in the inventories realised between 2000 and 2003. The last severe arbovirus epidemic of chikungunya (2005-2006), vectored by Ae. albopictus, which occurred in the area of the Indian Ocean emphasized the epidemiological role of this vector. So that, notifying its presence might help to target potential exotic arboviruses that could be introduced into the island. Furthermore this study allowed us to update the culicidae inventory, not only with this species, but also with the first observation of Culex sitiens in Europa and the confirmation of Ae. dufouri on this island. In order to determine the different species at the adult and/or at the larval stage, the authors propose keys of identification for mosquito species of the Eparse islands. All the described species on these islands are allochthonous species and some are being a nuisance. Sanitary recommendations together with biological and mechanical controls are proposed in order to reduce the nuisance and to prevent from any new introduction on these islands.

Genetic structure of the invasive pest Bemisia tabaci: evidence of limited but persistent genetic differentiation in glasshouse populations.

The geographic range of plant pests can be modified by the use of glasshouses. Bemisia tabaci, originating from warm to hot climates, has been shown to be a complex of distinct genetic groups with very limited gene flow. The genetic structure of this pest was studied in glasshouses in southern France, a region beyond the northern limit of its open-field development area in Europe. Seven microsatellite loci were scored in 22 populations sampled from various regions over 3 years. Two genetic groups were distinguished using a Bayesian clustering method and were assigned to the so-called biotypes B and Q using the gene sequence of cytochrome oxidase 1 (CO1). All but one population corresponded to biotype Q, even though only biotype B was previously reported. Despite the enclosed environment of glasshouses and their expected isolation due to low outdoor survival during the winter, only limited differentiation among biotype Q glasshouses was observed. A single sample site was notable for a decrease in expected heterozygosity and the mean number of alleles over the years. The lack of spatial genetic structure among biotype Q populations was indicative of a recent colonization event combined with large dispersal at all spatial scales. This migration pattern of biotype Q populations was further supported by additional CO1 sequences, since individuals from France, Asia and America exhibited 100% nucleotide identity. The evolution of genetic diversity observed in glasshouses in France is part of the worldwide invasion of biotype Q, which is discussed in light of human activities.

Geographic distribution and developmental sites of Aedes albopictus (Diptera: Culicidae) during a Chikungunya epidemic event.

Aedes albopictus is generally considered to have a low vectorial capacity because of its lack of host specificity. Nevertheless, it has been the sole vector of the Chikungunya virus in recent explosive epidemics on the islands of La Réunion and Mauritius. We report on investigations of the seasonal prevalence, container preferences, and geographic distribution of the species on La Réunion. Ae. albopictus showed strong ecological plasticity. In the warm wet season, small disposable containers were the principal urban breeding site, with 1939 positive containers in 750 houses. In the dry winter season, the species remained abundant throughout the island up to 800 m and was present to a maximum altitude of 1200 m. Natural containers were clearly important in this season, although productive sources were hard to find. The preferred natural developmental sites were bamboo stumps and rock holes, over 357 developmental sites observed in peri-urban and gully areas. Generalized logistic models indicated that the optimum sites contained clear water with high organic content and were situated in sites with moderate shade. Our data will provide input into the models of the epidemiology of the disease and design of vector control programs.