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Understanding how land-use change affects biodiversity is a fundamental step to develop effective conservation strategies in human-modified tropical landscapes. Here, we analyzed how land-use change through tropical small-scale agriculture affects endemic, exotic, and non-endemic native ant communities, focusing on vanilla landscapes in north-eastern Madagascar, a global biodiversity hotspot. First, we compared ant species richness and species composition across seven land-use types: old-growth forest, forest fragment, forest-derived vanilla agroforest, fallow-derived vanilla agroforest, woody fallow, herbaceous fallow, and rice paddy. Second, we assessed how environmental factors drive ant species richness in the agricultural matrix to identify management options that promote endemic and non-endemic native while controlling exotic ant species. We found that old-growth forest, forest fragment, and forest-derived vanilla agroforest supported the highest endemic ant species richness. Exotic ant species richness, by contrast, was lowest in old-growth forest but highest in herbaceous fallows, woody fallows, and rice paddy. Rice paddy had the lowest non-endemic native ant species richness. Ant species composition differed among land-use types, highlighting the uniqueness of old-growth forest in harboring endemic ant species which are more sensitive to disturbance. In the agricultural matrix, higher canopy closure and landscape forest cover were associated with an increase of endemic ant species richness but a decrease of exotic ant species richness. We conclude that preserving remnant forest fragments and promoting vanilla agroforests with a greater canopy closure in the agricultural matrix are important management strategies to complement the role of old-growth forests for endemic ant conservation in north-eastern Madagascar.

Aim: Elevational gradients are a useful approach to evaluate how environmental factors affect animal diversity. Decades of studies on the elevation-diversity gradient have revealed that this gradient varies greatly with taxa and geographic regions. One potential explanation for this may be the dependence of the relationship on landscape features. We explore the impact of fragmentation, habitat diversity and habitat amount on insect diversity (alpha and beta) and abundance along elevational gradients. We hypothesize that insect diversity and abundance will relate negatively with elevation, but positively with these landscape features. We also hypothesize that landscape features will interact in a way that the positive effect of a given variable on insect diversity may be offset by the others. Location: Reunion Island (Indian Ocean). Taxon: The insect order thrips (Thysanoptera). Methods: Insects were sampled along replicated elevational gradients, and at each sampling plot landscape features and abiotic variables were estimated within buffers surrounding the site. Insect alpha diversity was estimated using abundance-based rarefaction methods, whereas beta diversity was estimated calculating the "Local contributions to beta diversity" metric. The effect of elevation, rainfall, landscape features and their interactions was assessed on insect alpha and beta diversity and abundance during two consecutive seasons using linear mixed effects models. Results: We found that thrips alpha and beta diversity was negatively related with elevation, but the relationship varied between seasons and rainfall regimes. Among the different landscape features considered, we found that habitat amount had the strongest effect on diversity. The effect of habitat amount on diversity, however, was offset in areas of low habitat (or land cover) diversity. Main conclusions: Generalizing the factors that underlie the elevation diversity gradient has become a cornerstone in ecological theory because it can help to understand the impact of human activities on diversity. Here we show that taking landscape information into account may help to fulfil this objective because landscape effects co-vary with elevation with often intricate consequences for diversity.

In recent years, a significant increase in mosquito-borne diseases has been recorded worldwide. Faced with the limitations of existing methods for controlling the vector mosquito population, the development of attractants to bait traps and repellents to limit host-vector contacts could be promising and environmentally-friendly control strategies. The purpose of this study was to evaluate the effect of hydroxycoumarins and their alkyls derivatives against Aedes albopictus, the main vector of several arboviruses. Synthesis, bioassays and field trials were carried out in Madagascar. The results showed that 3, 4 and 6-hydroxycoumarins are attractive to this mosquito, 4-hydroxycoumarin being the most effective both in the laboratory and under field conditions. In addition, a good synergistic effect was found with octenol to attract mosquitoes and especially Ae. albopictus in comparison to other mosquito species living in sympatry. On the contrary, the 4-s-butoxycoumarin and 4-s-pentoxycoumarin derivatives had a repellent effect with the former showing the most significant effect. Further optimization of the dose and structure of these products will be carried out in order to maximize their utility for the control of Ae. albopictus and other mosquitoes.

On upland soils in tropical Africa, common production constraints of rice and maize on smallholder farms are poor soil fertility—resulting from soil erosion and nutrient depletion—and infestation by witchweeds (Striga spp.). In Madagascar where these crops are often grown in rotation, combining legume cover crops with no-till and crop residue mulching—labelled conservation agriculture (CA)—may address these problems. Previously, it was shown that CA practices contribute to steep reductions in Striga asiatica infection. In the current study, a 4-year field experiment was conducted to test, for the first time, the hypothesis that CA practices also contribute to crop yield and soil improvements under Striga-infested conditions. The conventional mono-crop rice–maize rotation practice, involving seasonal tillage and crop residue removal, was compared to three rice–maize rotation systems following CA practices, each with a different legume cover crop option: (1) two short-cycle annual species, cowpea (Vigna unguiculata) and mucuna (Mucuna pruriens); (2) a long-cycle annual, ricebean (Vigna umbellata); and (3) a perennial, stylosanthes (S. guianensis). Rice yields, as well as yield variability, generally increased by changing from the conventional to a CA practice, and maize yields were variable and low in particular under the CA practices. CA practices significantly reduced soil displacement by rainwater runoff and increased soil nitrogen and pH levels (0–20 cm depth), in particular with stylosanthes as cover crop, but did not result in a significant change in soil organic carbon concentration. Rice yields correlated negatively with Striga asiatica plant numbers in years with moderate infection levels. This is the first study that shows mixed outcomes from CA practices in tropical cereal rotation systems on degraded, Striga-infested soils, and subsequent entry points for system improvements. Suggested improvements include judicious cover crop management, complementary fertilizer applications and selection of competitive, resistant and adapted crop varieties.

International audience; Abstract Apis mellifera unicolor is a tropical honeybee endemic of Madagascar. Comprehensive knowledge about its mitochondrial and nuclear genetic diversity and structuration was our main purpose. Samples of worker bees were collected from 867 colonies in 76 sites in Madagascar and 1 reference population in South Africa. PCR-restriction fragment length polymorphism (RFLP) and sequencing were used to reveal variability in the COI–COII mtDNA region. Seventeen microsatellite loci were used for studying the nuclear diversity. Three PCR-RFLP profiles were observed, among which 99.4 % belonged to A1 haplotype, 0.2 % to a new A haplotype, and 0.4 % to A4 haplotype. In microsatellite analysis, moderate genetic diversity values were found for Madagascar, together with low mean number of alleles ranging from 2.47 to 3.88 compared to South Africa. Bayesian clustering assignment methods and principal component analysis (PCA) separated populations into two genetic clusters which matched with geographic areas. Several hypotheses are discussed regarding to the low genetic diversity of A. m. unicolor in its native range.

Objective: To evaluate the potential efficacy of seed extracts of Annona squamosa and Annona muricata used as natural insecticides to control adult and larvae of the vectors Aedes albopictus and Culex quinquefasciatus under laboratory conditions. Methods: Aqueous and oil extracts of the two plants were prepared from dried seeds. Preliminary identifications of the chemical components of each seed extracts were performed using microreactional and GCP techniques. Larvae and adults of Aedes albopictus and Culex quinquefasciatus were collected from the breeding sites in coastal and highlands regions of Madagascar. WHO standardized tests of susceptibility for larvae and imaginal stage of mosquitoes were realized to determine mortality and LC50 of mosquitoes. Results: Chemical identifications showed that these extracts contain alkaloids and flavonoids compounds that probably confer their biological insecticidal proprieties. CPG analysis showed also the presence of various fatty acids. On adult mosquitoes, significant insecticidal effects were observed with both aqueous and oil extracts of the two plant seeds compared to mortality induced by deltamethrin, an insecticide used as reference. Extracts of Annona muricata induced high mortality rate to both species of mosquito compared to extracts of Annona squamosa at all concentrations tested. The LC50 of seed extracts ranged from 1% to 5% for adults and 0.5% to 1% for larvae. Conclusions: The seed extracts of these two plants may be used as mosquito controlling agents and offer a new approach to a less costly, practical and environmentally friendly control of vector borne diseases. Peer reviewer

Symbiotic bacteria are known to play important roles in the biology of insects, but the current knowledge of bacterial communities associated with mosquitoes is very limited and consequently their contribution to host behaviors is mostly unknown. In this study, we explored the composition and diversity of mosquito-associated bacteria in relation with mosquitoes' habitats. Wild Aedes albopictus and Aedes aegypti were collected in three different geographic regions of Madagascar. Culturing methods and denaturing gradient gel electrophoresis (DGGE) and sequencing of the rrs amplicons revealed that Proteobacteria and Firmicutes were the major phyla. Isolated bacterial genera were dominated by Bacillus, followed by Acinetobacter, Agrobacterium and Enterobacter. Common DGGE bands belonged to Acinetobacter, Asaia, Delftia, Pseudomonas, Enterobacteriaceae and an uncultured Gammaproteobacterium. Double infection by maternally inherited Wolbachia pipientis prevailed in 98% of males (n=272) and 99% of females (n=413); few individuals were found to be monoinfected with Wolbachia wAlbB strain. Bacterial diversity (Shannon-Weaver and Simpson indices) differed significantly per habitat whereas evenness (Pielou index) was similar. Overall, the bacterial composition and diversity were influenced both by the sex of individuals and by the environment inhabited by the mosquitoes; the latter might be related to both the vegetation and the animal host populations that Aedes used as food sources.

International audience; Varroa destructor is a major pest in world beekeeping. It was first detected in Madagascar in 2010 on the endemic honeybee Apis mellifera unicolor. To evaluate V. destructor spread dynamics in Madagascar a global survey was conducted in 2011–2012. A total of 695 colonies from 30 districts were inspected for the presence of mites. 2 years after its introduction, nine districts were found infested. Varroa destructor spread was relatively slow compared to other countries with a maximum progression of 40 km per year, the five newly infested districts being located next to the first infested ones. The incidence of mite infestation was also investigated by monitoring 73 colonies from five apiaries during 1 year (2011–2012). Sixty percent of local colony mortality was recorded after 1 year of survey. Varroa destructor strain determination was done by partial sequencing of the cytochrome oxidase I gene of 13 phoretic mites sampled in five districts. A single V. destructor mitochondrial haplotype was detected, the Korean type, also present in the closest African countries. A global pathogen survey was also conducted on the colonies inspected for mite presence. The greater wax moth, Galleria mellonella has been found in all colonies all over the country. Two other pathogens and morphological abnormalities in workers, such as deformed wings, were found associated with only V. destructor presence. A prevention management plan must be implemented to delimit mite spread across the island.

Background In the past ten years, the Indian Ocean region has been the theatre of severe epidemics of chikungunya and dengue. These outbreaks coincided with a high increase in populations of Aedes albopictus that outcompete its sister taxon Aedes aegypti in most islands sampled. The objective of this work was to update the entomological survey of the two Aedes species in the island of Madagascar which has to face these arboviroses. Methods The sampling of Aedes mosquitoes was conducted during two years, from October 2007 to October 2009, in fifteen localities from eight regions of contrasting climates. Captured adults were identified immediately whereas immature stages were bred until adult stage for determination. Phylogenetic analysis was performed using two mtDNA genes, COI and ND5 and trees were constructed by the maximum likelihood (ML) method with the gene time reversible (GTR) model. Experimental infections with the chikungunya virus strain 06.21 at a titer of 107.5 pfu/mL were performed to evaluate the vector competence of field-collected mosquitoes. Disseminated infection rates were measured fourteen days after infection by immunofluorescence assay performed on head squashes. Results The species Aedes aegypti was detected in only six sites in native forests and natural reserves. In contrast, the species Aedes albopictus was found in 13 out of the 15 sites sampled. Breeding sites were mostly found in man-made environments such as discarded containers, used tires, abandoned buckets, coconuts, and bamboo cuts. Linear regression models showed that the abundance of Ae. albopictus was significantly influenced by the sampling region (F = 62.00, p < 2.2 × 10-16) and period (F = 36.22, p = 2.548 × 10-13), that are associated with ecological and climate variations. Phylogenetic analysis of the invasive Ae. albopictus distinguished haplotypes from South Asia and South America from those of Madagascar, but the markers used were not discriminant enough to discern Malagasy populations. The experimental oral infection method showed that six Ae. albopictus populations exhibited high dissemination infection rates for chikungunya virus ranging from 98 to 100%. Conclusion In Madagascar, Ae. albopictus has extended its geographical distribution whereas, Ae. aegypti has become rare, contrasting with what was previously observed. Changes are predominantly driven by human activities and the rainfall regime that provide suitable breeding sites for the highly anthropophilic mosquito Ae. albopictus. Moreover, these populations were found to be highly susceptible to chikungunya virus. In the light of this study, Ae. albopictus may have been involved in the recent outbreaks of chikungunya and dengue epidemics in Madagascar, and consequently, control measures should be promoted to limit its current expansion.

In Madagascar, growing demand for rice and resulting increased pressure on inundated lands has favoured the cultivation of upland rain-fed rice on hill slopes. Due to the instability of the ecosystem, this type of agriculture cannot meet the objectives of both sustainability and high yields if conventional tillage is used, particularly due to high erosion risk. Direct seeding, mulch-based cropping (DMC) systems have opened new prospects for upland rice.

Symbiotic bacteria are known to play important roles in the biology of insects, but the current knowledge of bacterial communities associated with mosquitoes is very limited and consequently their contribution to host behaviors is mostly unknown. In this study, we explored the composition and diversity of mosquito-associated bacteria in relation with mosquitoes' habitats. Wild Aedes albopictus and Aedes aegypti were collected in three different geographic regions of Madagascar. Culturing methods and denaturing gradient gel electrophoresis (DGGE) and sequencing of the rrs amplicons revealed that Proteobacteria and Firmicutes were the major phyla. Isolated bacterial genera were dominated by Bacillus, followed by Acinetobacter, Agrobacterium and Enterobacter. Common DGGE bands belonged to Acinetobacter, Asaia, Delftia, Pseudomonas, Enterobacteriaceae and an uncultured Gammaproteobacterium. Double infection by maternally inherited Wolbachia pipientis prevailed in 98% of males (n=272) and 99% of females (n=413); few individuals were found to be monoinfected with Wolbachia wAlbB strain. Bacterial diversity (Shannon-Weaver and Simpson indices) differed significantly per habitat whereas evenness (Pielou index) was similar. Overall, the bacterial composition and diversity were influenced both by the sex of individuals and by the environment inhabited by the mosquitoes; the latter might be related to both the vegetation and the animal host populations that Aedes used as food sources.