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.

Genetic mapping of a major dominant gene for resistance to Ralstonia solanacearum in eggplant.

Resistance of eggplant against Ralstonia solanacearum phylotype I strains was assessed in a F(6) population of recombinant inbred lines (RILs) derived from a intra-specific cross between S. melongena MM738 (susceptible) and AG91-25 (resistant). Resistance traits were determined as disease score, percentage of wilted plants, and stem-based bacterial colonization index, as assessed in greenhouse experiments conducted in Réunion Island, France. The AG91-25 resistance was highly efficient toward strains CMR134, PSS366 and GMI1000, but only partial toward the highly virulent strain PSS4. The partial resistance found against PSS4 was overcome under high inoculation pressure, with heritability estimates from 0.28 to 0.53, depending on the traits and season. A genetic map was built with 119 AFLP, SSR and SRAP markers positioned on 18 linkage groups (LG), for a total length of 884 cM, and used for quantitative trait loci (QTL) analysis. A major dominant gene, named ERs1, controlled the resistance to strains CMR134, PSS366, and GMI1000. Against strain PSS4, this gene was not detected, but a significant QTL involved in delay of disease progress was detected on another LG. The possible use of the major resistance gene ERs1 in marker-assisted selection and the prospects offered for academic studies of a possible gene for gene system controlling resistance to bacterial wilt in solanaceous plants are discussed.

Influence of risk factors on renal leptospiral load in naturally infected wild black rats.

In 2009, a survey was conducted in Reunion Island to determine the renal leptospiral load in black rats trapped in the field. The concentration of leptospires in kidney tissue was calculated using qPCR. Our results showed high inter-individual variations of renal bacterial load in naturally infected black rats (mean=8.27 ± 4.72 log-genome copies per mg kidney tissue). The objective of this study was to model the renal leptospiral load in 50 naturally infected black rats as a function of sex, age, and weight. Statistical analysis by sex showed that, in naturally infected males, the renal leptospiral load was correlated with weight (p-value=0.032). Moreover, our model showed that weight and sex were significant explanatory variables for the renal leptospiral load in naturally infected young black rats (R(2)=0.953). Laboratory experimentation could not replicate naturally acquired infection, but field studies also present many limitations. Our study is the first attempt to explain individual variations in the renal leptospiral load in naturally infected reservoir animals but further research is needed.

Species diversity within a community of the curcurbit fruit flies Bactrocera cucurbitae, Dacus ciliatus, and Dacus demmerezi roosting in corn borders near cucurbit production areas of Reunion Island.

In order to better control fruit flies (Diptera: Tephritidae) attacking Cucurbitaceae on Reunion Island (21°6 S/ 55°36 E), biological characteristics (seasonal fluctuation, relative abundance, sex ratio) of communities roosting in corn borders were investigated. The study was conducted in austral summer across a range of altitudes (750-1150 m) corresponding to the main areas of cucurbit cropping. Living adults were recorded roosting on corn planted within or around cucurbit fields. Results showed a high variability in seasonal fluctuation of populations according to local conditions. Bactrocera Cucurbitae (Coquillett) (Diptera: Tephritidae) was the least abundant species (27%) compared to Dacus ciliatus Loew (36%) and Dacus demmerezi Bezzi (37%). Relative abundance of B. Cucurbitae was lowest (< 18%) in high altitude sites (above 1000 m), where D. demmerezi was the most prevalent species (> 56%). Dacus ciliatus showed variable relative abundance (from 18 to 51%) depending on the experimental design (varying in location and in year). Sex ratio was also very variable from one species to another and from one experimental design to another.

So near and yet so far: the specific case of Ralstonia Solanacearum populations from Côte d'Ivoire in Africa.

The genetic and phenotypic diversity of Côte d'Ivoire Ralstonia solanacearum strains was assessed on a 168-strain collection sampled on Solanaceae both in the southern lowlands and western highlands. Phylotypes I, II, and III were prevalent, though at unexpected frequencies. Phylotype I strains (87.5%) were genetically diverse and overrepresented in all agroecological areas, including highlands (AEZ III). Phylotype II strains (10.7%) only belonged to one tropical lowland-adapted broad host range lineage (IIA-35), whereas no highland-adapted potato brown rot (IIB-1) or Moko strains were detected. African phylotype III strains were rare (1.8%). They originated from a single Burkina Faso lineage (III-23) and were only found in lowlands. Three phylotype I strains were found harboring pRSC35, a plasmid identified in phylotype III strains in Cameroon. From pathogenicity tests performed on commercial varieties and tomato/eggplant/pepper references, the virulence diversity observed was high, with five pathoprofiles described. Eggplant accessions MM152 and EG203 and tomato HW7996 displayed the largest resistance spectrum and highest level. Two highly virulent phylotype I strains were able to bypass resistance of HW7996 and the eggplant reference AG91-25. Collectively, these points lead to the conclusion that the situation in Côte d'Ivoire is specific towards other African countries, and specifically from the Cameroon reference, and that within phylotype I can exist a high virulence diversity. This calls for similar studies in neighboring West African countries, linking R. solanacearum pathogen genetic diversity to strain virulence at the regional level, for the rationalization of regional resistance deployment strategies and future resistance durability studies.

MultiLocus Sequence Analysis- and Amplified Fragment Length Polymorphism-based characterization of xanthomonads associated with bacterial spot of tomato and pepper and their relatedness to Xanthomonas species.

MultiLocus Sequence Analysis (MLSA) and Amplified Fragment Length Polymorphism (AFLP) were used to measure the genetic relatedness of a comprehensive collection of xanthomonads pathogenic to solaneous hosts to Xanthomonas species. The MLSA scheme was based on partial sequences of four housekeeping genes (atpD, dnaK, efp and gyrB). Globally, MLSA data unambiguously identified strains causing bacterial spot of tomato and pepper at the species level and was consistent with AFLP data. Genetic distances derived from both techniques showed a close relatedness of (i) X. euvesicatoria, X. perforans and X. alfalfae and (ii) X. gardneri and X. cynarae. Maximum likelihood tree topologies derived from each gene portion and the concatenated data set for species in the X. campestris 16S rRNA core (i.e. the species cluster comprising all strains causing bacterial spot of tomato and pepper) were not congruent, consistent with the detection of several putative recombination events in our data sets by several recombination search algorithms. One recombinant region in atpD was identified in most strains of X. euvesicatoria including the type strain.

Phylogeny and population structure of brown rot- and Moko disease-causing strains of Ralstonia solanacearum phylotype II.

The ancient soilborne plant vascular pathogen Ralstonia solanacearum has evolved and adapted to cause severe damage in an unusually wide range of plants. In order to better describe and understand these adaptations, strains with very similar lifestyles and host specializations are grouped into ecotypes. We used comparative genomic hybridization (CGH) to investigate three particular ecotypes in the American phylotype II group: (i) brown rot strains from phylotypes IIB-1 and IIB-2, historically known as race 3 biovar 2 and clonal; (ii) new pathogenic variants from phylotype IIB-4NPB that lack pathogenicity for banana but can infect many other plant species; and (iii) Moko disease-causing strains from phylotypes IIB-3, IIB-4, and IIA-6, historically known as race 2, that cause wilt on banana, plantain, and Heliconia spp. We compared the genomes of 72 R. solanacearum strains, mainly from the three major ecotypes of phylotype II, using a newly developed pangenomic microarray to decipher their population structure and gain clues about the epidemiology of these ecotypes. Strain phylogeny and population structure were reconstructed. The results revealed a phylogeographic structure within brown rot strains, allowing us to distinguish European outbreak strains of Andean and African origins. The pangenomic CGH data also demonstrated that Moko ecotype IIB-4 is phylogenetically distinct from the emerging IIB-4NPB strains. These findings improved our understanding of the epidemiology of important ecotypes in phylotype II and will be useful for evolutionary analyses and the development of new DNA-based diagnostic tools.

Insertion sequence- and tandem repeat-based genotyping techniques for Xanthomonas citri pv. mangiferaeindicae.

Molecular fingerprinting techniques that have the potential to identify or subtype bacteria at the strain level are needed for improving diagnosis and understanding of the epidemiology of pathogens such as Xanthomonas citri pv. mangiferaeindicae, which causes mango bacterial canker disease. We developed a ligation-mediated polymerase chain reaction targeting the IS1595 insertion sequence as a means to differentiate pv. mangiferaeindicae from the closely related pv. anacardii (responsible for cashew bacterial spot), which has the potential to infect mango but not to cause significant disease. This technique produced weakly polymorphic fingerprints composed of ≈70 amplified fragments per strain for a worldwide collection of X. citri pv. mangiferaeindicae but produced no or very weak amplification for pv. anacardii strains. Together, 12 tandem repeat markers were able to subtype X. citri pv. mangiferaeindicae at the strain level, distinguishing 231 haplotypes from a worldwide collection of 299 strains. Multilocus variable number of tandem repeats analysis (MLVA), IS1595-ligation-mediated polymerase chain reaction, and amplified fragment length polymorphism showed differences in discriminatory power and were congruent in describing the diversity of this strain collection, suggesting low levels of recombination. The potential of the MLVA scheme for molecular epidemiology studies of X. citri pv. mangiferaeindicae is discussed.

Bacterial wilt resistance in tomato, pepper, and eggplant: genetic resources respond to diverse strains in the Ralstonia solanacearum species complex.

Bacterial wilt, caused by strains belonging to the Ralstonia solanacearum species complex, inflicts severe economic losses in many crops worldwide. Host resistance remains the most effective control strategy against this disease. However, wilt resistance is often overcome due to the considerable variation among pathogen strains. To help breeders circumvent this problem, we assembled a worldwide collection of 30 accessions of tomato, eggplant and pepper (Core-TEP), most of which are commonly used as sources of resistance to R. solanacearum or for mapping quantitative trait loci. The Core-TEP lines were challenged with a core collection of 12 pathogen strains (Core-Rs2) representing the phylogenetic diversity of R. solanacearum. We observed six interaction phenotypes, from highly susceptible to highly resistant. Intermediate phenotypes resulted from the plants' ability to tolerate latent infections (i.e., bacterial colonization of vascular elements with limited or no wilting). The Core-Rs2 strains partitioned into three pathotypes on pepper accessions, five on tomato, and six on eggplant. A "pathoprofile" concept was developed to characterize the strain clusters, which displayed six virulence patterns on the whole set of Core-TEP host accessions. Neither pathotypes nor pathoprofiles were phylotype specific. Pathoprofiles with high aggressiveness were mainly found in strains from phylotypes I, IIB, and III. One pathoprofile included a strain that overcame almost all resistance sources.

Pseudomonas aeruginosa outbreak linked to mineral water bottles in a neonatal intensive care unit: fast typing by use of high-resolution melting analysis of a variable-number tandem-repeat locus.

Pseudomonas aeruginosa is an opportunistic pathogen that causes nosocomial infections in intensive care units. Determining a system of typing that is discriminatory is essential for epidemiological surveillance of P. aeruginosa. We developed a method for the typing of Pseudomonas aeruginosa, namely, multiple-locus variable-number tandem-repeat (VNTR) typing with high-resolution melting analysis (HRMA). The technology was used to genotype a collection of 43 environmental and clinical strains isolated during an outbreak in a neonatal intensive care unit (NICU) that we report. Nineteen strains isolated in other departments or outside the hospital were also tested. The genetic diversity of this collection was determined using VNTR-HRMA, with amplified fragment length polymorphism (AFLP) analysis as a reference. Twenty-five and 28 genotypes were identified, respectively, and both techniques produced congruent data. VNTR-HRMA established clonal relationships between the strains of P. aeruginosa isolated during the outbreak in the NICU and proved, for the first time, the role of mineral water as the inoculum source. VNTR typing with one primer pair in association with HRMA is highly reproducible and discriminative, easily portable among laboratories, fast, and inexpensive, and it demonstrated excellent typeability in this study. VNTR-HRMA represents a promising tool for the molecular surveillance of P. aeruginosa and perhaps for molecular epidemiologic analysis of other hospital infections.