Adaptation of genetically monomorphic bacteria: evolution of copper resistance through multiple horizontal gene transfers of complex and versatile mobile genetic elements.

Copper-based antimicrobial compounds are widely used to control plant bacterial pathogens. Pathogens have adapted in response to this selective pressure. Xanthomonas citri pv. citri, a major citrus pathogen causing Asiatic citrus canker, was first reported to carry plasmid-encoded copper resistance in Argentina. This phenotype was conferred by the copLAB gene system. The emergence of resistant strains has since been reported in Réunion and Martinique. Using microsatellite-based genotyping and copLAB PCR, we demonstrated that the genetic structure of the copper-resistant strains from these three regions was made up of two distant clusters and varied for the detection of copLAB amplicons. In order to investigate this pattern more closely, we sequenced six copper-resistant X. citri pv. citri strains from Argentina, Martinique and Réunion, together with reference copper-resistant Xanthomonas and Stenotrophomonas strains using long-read sequencing technology. Genes involved in copper resistance were found to be strain dependent with the novel identification in X. citri pv. citri of copABCD and a cus heavy metal efflux resistance-nodulation-division system. The genes providing the adaptive trait were part of a mobile genetic element similar to Tn3-like transposons and included in a conjugative plasmid. This indicates the system's great versatility. The mining of all available bacterial genomes suggested that, within the bacterial community, the spread of copper resistance associated with mobile elements and their plasmid environments was primarily restricted to the Xanthomonadaceae family.

Extension of the sorting instructions for household plastic packaging and changes in exposure to bioaerosols at materials recovery facilities.

The aim of this study was to assess how extending the sorting instructions for plastic packaging would affect the exposure of workers working at materials recovery facility (MRF) to dust, endotoxins, fungi and bacteria, taking into consideration other factors that could have an influence on this exposure. Personal sampling was carried out at four MRFs during six sampling campaigns at each facility, both in sorting rooms and when the workers were involved in "mobile tasks" away from the rooms. The data was analysed by describing the extension of sorting instructions both using a qualitative variable (after vs before) and using data for the pots and trays recycling stream, including or excluding plastic film. Overall, before the extension of the sorting guidelines, the geometric mean of personal exposure levels in sorting rooms was 0.3mg/m(3) for dust, 27.7 EU/m(3) for endotoxins, 13,000 CFU/m(3) for fungi and 1800 CFU/m(3) for bacteria. When workers were involved in mobile tasks away from the rooms, these averages were 0.5mg/m(3), 25.7 EU/m(3), 28,000 CFU/m(3) and 5100 CFU/m(3) respectively.The application by households of instructions to include pots, trays and film with other recyclable plastic packaging led to an increase in exposure to endotoxins, fungi and bacteria at MRFs. For an increase of 0.5 kg per inhabitant per year in the pots, trays and film recycling stream, exposure in sorting rooms rose by a factor of 1.4-2.2, depending on the biological agent. Exposure during mobile tasks increased by a factor of 3.0-3.6. The age of the waste amplified the effect of the extension of sorting instructions on exposure to fungi, bacteria and endotoxins. Factors that had a significant influence on the exposure of workers to dust and/or bioaerosols included the presence of paper, newspapers and magazines in the sorted waste, the order in which incoming waste was treated and the quality of the ventilation system in the sorting rooms. The levels of exposure observed in this study highlight the need to implement appropriate preventive measures against bioaerosols at MRFs for dry waste. There are grounds to justify these preventive measures, both inside sorting rooms and for the MRF as a whole, regardless of whether the decision to extend sorting instructions for household plastic waste is adopted.

Rapid increase in dispersal during range expansion in the invasive ladybird Harmonia axyridis.

The evolutionary trajectories associated with demographic, genetic and spatial disequilibrium have become an issue of growing interest in population biology. Invasive species provide unique opportunities to explore the impact of recent range expansion on life-history traits, making it possible to test for a spatial arrangement of dispersal abilities along the expanding range, in particular. We carried out controlled experiments in laboratory conditions to test the hypothesis of an increase in dispersal capacity with range expansion in Harmonia axyridis, a ladybird that has been invading Europe since 2001. We found a marked increase in the flight speed of the insects from the core to the front of the invasion range in two independent sampling transects. By contrast, we found that two other traits associated with dispersal (endurance and motivation to fly off) did not follow the same spatial gradient. Our results provide a striking illustration of the way in which predictable directional genetic changes may occur rapidly for some traits associated with dispersal during biological invasions. We discuss the consequences of our results for invasion dynamics and the evolutionary outcomes of spatially expanding populations.

Role of propagule pressure in colonization success: disentangling the relative importance of demographic, genetic and habitat effects.

High propagule pressure is arguably the only consistent predictor of colonization success. More individuals enhance colonization success because they aid in overcoming demographic consequences of small population size (e.g. stochasticity and Allee effects). The number of founders can also have direct genetic effects: with fewer individuals, more inbreeding and thus inbreeding depression will occur, whereas more individuals typically harbour greater genetic variation. Thus, the demographic and genetic components of propagule pressure are interrelated, making it difficult to understand which mechanisms are most important in determining colonization success. We experimentally disentangled the demographic and genetic components of propagule pressure by manipulating the number of founders (fewer or more), and genetic background (inbred or outbred) of individuals released in a series of three complementary experiments. We used Bemisia whiteflies and released them onto either their natal host (benign) or a novel host (challenging). Our experiments revealed that having more founding individuals and those individuals being outbred both increased the number of adults produced, but that only genetic background consistently shaped net reproductive rate of experimental populations. Environment was also important and interacted with propagule size to determine the number of adults produced. Quality of the environment interacted also with genetic background to determine establishment success, with a more pronounced effect of inbreeding depression in harsh environments. This interaction did not hold for the net reproductive rate. These data show that the positive effect of propagule pressure on founding success can be driven as much by underlying genetic processes as by demographics. Genetic effects can be immediate and have sizable effects on fitness.

Investigating the genetic load of an emblematic invasive species: the case of the invasive harlequin ladybird Harmonia axyridis.

Introduction events can lead to admixture between genetically differentiated populations and bottlenecks in population size. These processes can alter the adaptive potential of invasive species by shaping genetic variation, but more importantly, they can also directly affect mean population fitness either increasing it or decreasing it. Which outcome is observed depends on the structure of the genetic load of the species. The ladybird Harmonia axyridis is a good example of invasive species where introduced populations have gone through admixture and bottleneck events. We used laboratory experiments to manipulate the relatedness among H. axyridis parental individuals to assess the possibility for heterosis or outbreeding depression in F1 generation offspring for two traits related to fitness (lifetime performance and generation time). We found that inter-populations crosses had no major impact on the lifetime performance of the offspring produced by individuals from either native or invasive populations. Significant outbreeding depression was observed only for crosses between native populations for generation time. The absence of observed heterosis is indicative of a low occurrence of fixed deleterious mutations within both the native and invasive populations of H. axyridis. The observed deterioration of fitness in native inter-population crosses most likely results from genetic incompatibilities between native genomic backgrounds. We discuss the implications of these results for the structure of genetic load in H. axyridis in the light of the available information regarding the introduction history of this species.

Experimental evidence for the phenotypic impact of admixture between wild and biocontrol Asian ladybird (Harmonia axyridis) involved in the European invasion.

Hybridization can fuel evolutionary processes during biological invasions. The harlequin ladybird Harmonia axyridis has long been used as a biocontrol agent before the species became invasive worldwide. Previous analysis based on microsatellite data has shown that European invasive populations bear traces of admixture between an eastern North American source, which is at the origin of the worldwide invasion, and biocontrol strains used in Europe. In this study, we tested the hypothesis that this early admixture event may have fostered the European invasion by impacting on the phenotypes of wild European populations. Mean life history traits of experimental F(1) hybrids are compared with pure parental sources and wild European crosses. Our results reveal a biased impact whereby North American beetles benefitted from being admixed with European biocontrol strains. Resemblance between experimental hybrids and wild European invasive crosses further suggests a long-lasting effect of admixture that may still be at work and fostering invasiveness.

Hybridization and invasiveness in the freshwater snail Melanoides tuberculata: hybrid vigour is more important than increase in genetic variance.

Many invasive taxa are hybrids, but how hybridization boosts the invasive process remains poorly known. We address this question in the clonal freshwater snail Melanoides tuberculata from Martinique, using three parental and two hybrid lines. We combine an extensive field survey (1990-2003) and a quantitative genetic experiment to show that hybrid lines have outcompeted their parents in natural habitats, and that this increased invasiveness co-occurred with pronounced shifts in life-history traits, such as growth, fecundity and juvenile size. Given the little time between hybrid creation and sampling, and the moderate standing genetic variance for life-history traits in hybrids, we show that some of the observed trait changes between parents and hybrids were unlikely to arise only by continuous selection. We therefore suggest that a large part of hybrid advantage stems from immediate heterosis upon hybridization.

A molecular phylogeography approach to biological invasions of the New World by parthenogenetic Thiarid snails.

The parthenogenetic snail Melanoides tuberculata, present in tropical fresh waters of most of the Old World before 1950, has now invaded the Neotropical area. The phylogeography of this snail was studied to evaluate the pathways and number of such invasions. Because of parthenogenetic reproduction, individuals are structured into genetical clones. Within populations from both the original and invaded areas, several morphologically distinct clones (referred to as morphs) often coexist but the amount of genetic divergence among morphs is unknown. Individuals from 27 morphs and 40 populations world-wide were sequenced at two mitochondrial genes (12S and 16S). Our phylogenetic reconstruction suggests that (i) most of the morphological variation observed in the New World predates invasion, (ii) at least six independent introductions have occurred, and (iii) invasive clones are found throughout most of the phylogenetic tree and do not come from a particular region of the area of origin. Two ideas are discussed in the light of these results. The first lies with the specificities of parthenogenesis in an invasion context. While in sexual species, independently introduced populations eventually merge into a single invasive population, in a parthenogenetic species independently introduced clones have distinct invasion dynamics and possibly exclude each other. Second, although repeated invasions in Melanoides may have an impact on indigenous molluscan faunas, their most likely effect is the world-wide homogenization of the invasive taxon itself.

Chronological age and crystallized intelligence of people with intellectual disability.

The influence of chronological age (CA) and fluid intelligence on the crystallized intelligence level of people with intellectual disability was studied in a group of 102 participants aged between 6 and 20 years. The results, which were based on their performance in 12 fluid and crystallized intelligence markers, indicate that the fluid intelligence factor and CA explain an important fraction of crystallized intelligence factor variance (43% and 21%, respectively). This finding provides support for the hypothesis that CA-related experience exerts a significant effect on the crystallized component of intelligence in people with intellectual disability.

Chronological age and performance of persons with mental retardation on verbal subtests of the Wechsler Intelligence Scale for Children--Revised, French Version.

The Influence of chronological age on verbal intelligence of persons with metal retardation was studied using the French version of the Wechsler Intelligence Scale for Children--Revised. Participants were 1,101 children and adolescents divided into two groups according to the severity of metal retardation. The first group included 551 participants with moderate mental retardation, the second 550 participants with mild mental retardation. Analyses indicated a significant association between chronological age and raw scores on the four Verbal Comprehension subtests, even with performance on Perceptual Organization and Freedom from Distractibility subtests held constant. This finding suggests the relationship between chronological age and verbal intelligence is valid for persons with moderate mental retardation as well as for those with mild metal retardation.

Chronological age and receptive vocabulary of persons with Down syndrome.

The Test de Vocabulaire en Images (the French version of the Peabody Picture Vocabulary Test) and the Columbia Mental Maturity Scale were administered to 29 participants with Down syndrome aged 6.1 yr. to 13.0 yr. Analysis indicated a significant association between chronological age and the Test de Vocabulaire en Images mental age. This finding seems to suggest that the relationship between chronological age and the receptive vocabulary of persons with mental retardation is not specific to etiology.

Chronological age and Peabody Picture Vocabulary Test performance of persons with mental retardation: new data.

The Test de Vocabulaire en Images (the French version of the Peabody Picture Vocabulary Test) and the Columbia Mental Maturity Scale were administered to 44 persons with mental retardation who were aged 6.8 yr. to 18.2 yr. Analysis indicated a significant influence of chronological age on the Test de Vocabulaire en Images Mental Age. This influence of chronological age probably explains why receptive vocabulary tests consistently overestimate the IQ of persons with mental retardation.

Evaluation of toy variation on engagement in a leisure activity of two children with profound multiple handicaps.

We conducted two studies to examine whether satiation occurs during leisure activities for children with profound multiple handicaps and whether it can be prevented by the alternate use of a sufficient number of different toys. The two studies were done in a training program in which the use of a switch connected to battery-operated toys was taught to two children with profound multiple handicaps. In Study I (Child 1), two experimental conditions were compared: variation (use of 5 toys in the same learning session) and identity (use of only one of these toys throughout the session). The variation condition produced higher and more stable performance than the identity condition, which yielded a progressive decrease in the appearance of the target behavior. In Study 2 (Child 2), satiation appeared from one session to another and the rate of responding increased when a new toy was presented. Some practical and theoretical implications are discussed.

Overestimation of mentally retarded persons' IQs using the PPVT: a re-analysis and some implications for future research.

Numerous validity studies have shown that the PPVT consistently overestimates mentally retarded persons' IQs. One possible interpretation is that this phenomenon is an outcome of the dissociation between their cognitive level and their experience. Indeed, compared to intellectually average subjects of the same mental age, they have had more learning opportunities, simply because they have lived longer. In order to validate this hypothesis, the French version of the PPVT, Raven's Coloured Progressive Matrices (RCPM), and the 1966 version of the Binet-Simon were administered to 90 subjects of various chronological ages matched on mental age (30 nonretarded 5 year olds and two groups of 30 retarded subjects aged 10 and 16 years, respectively). The results indicate that CA exerts a strong effect on vocabulary, but not on RCPM performance. The research implications of this finding are discussed.

Diagnostic value of a synthetic peptide derived from Echinococcus granulosus recombinant protein.

A specific monoclonal antibody (MAb; EG 02 154/12) directed against a protein epitope of Echinococcus granulosus antigen 5 was used to screen a cDNA library constructed from E. granulosus protoscoleces RNA. One clone designated Eg14 was selected and shown to code for an amino acid sequence partially homologous to that of the clone Eg6 previously identified with the same MAb. Hydrophobic cluster analysis showed that both recombinant antigens may adopt a similar alpha-helical organization and share a common conformational epitope. A synthetic peptide (89-122) mimicking the conformational site of Eg6 and Eg14 was constructed and demonstrated to be able to inhibit binding of the MAb and human hydatid sera to the Eg6 fusion protein (FP6) or to native hydatid antigens. To assess the diagnostic value of the peptide 89-122, we tested sera from patients infected with different parasites for their antibody reactivity with this peptide in ELISA. A high binding sensitivity and specificity of IgG-A-M antibodies were obtained with E. granulosus-infected patient sera. Moreover, the peptide 89-122 was found to be specifically recognized by IgE antibodies from patients with hydatid disease. These results indicate the particular interest of this synthetic peptide as a standardized antigen in diagnosis and treatment surveillance of hydatidosis.

Molecular cloning of an Echinococcus granulosus protein expressing an immunogenic epitope of antigen 5.

cDNA was synthesized from RNA extracted from Echinococcus granulosus protoscoleces and cloned in the lambda gt11 expression vector. A pool of 5 E. granulosus patient sera was used to screen the library and allowed the selection of 13 clones. Ten of these were shown to be identical, among which clone 6 (Eg6) was chosen for further analysis. The nucleotide sequence (456-bp) presented an entire open reading frame coding for 152 amino acids. The fusion protein (FP6) was recognized by a mouse monoclonal antibody (EG 02 154/12) specific for E. granulosus antigen 5. Moreover, the presence of antibodies to FP6 seemed to be correlated to the ability of sera from hydatidosis patients to immunoprecipitate antigen 5. These results indicate that the cloned protein could be used as a standardized antigen for the diagnosis of hydatidosis.

Use of a monoclonal antibody specific for a protein epitope of Echinococcus granulosus antigen 5 in a competitive antibody radioimmunoassay for diagnosis of hydatid disease.

A monoclonal antibody (mAb) designated as EG 02 154/12, specific for the major antigen (antigen 5) of Echinococcus granulosus was produced, and used to study the binding sites recognized by anti-antigen 5 antibodies from patients with hydatid disease. The nature of the target epitope was partially characterized. The antibody reactivity was analyzed towards sheep hydatid fluid antigens (SHF Ag) using ELISA, immunoelectrophoresis (IEP), Western blotting (WB), and immunoprecipitation (IP). In IEP, EG 02 154/12 mAb gave a single precipitin of Ag 5. The mAb and human hydatid patient sera recognized a major antigen of 64 kDa, in SHF Ag analyzed in non-reducing conditions. Both types of antibodies revealed two components of 37 and 22 kDa in reducing conditions. Deglycosylation and delipidation of SHF Ag did not affect the mAb binding. These results, together with the observation of mAb binding to in vitro translation products from protoscoleces messenger RNA, suggest the protein nature of the epitope recognized on the antigen 5. Using competitive antibody radioimmunoassay (CRIA), a competition between this mAb and hydatid patient sera, for the same epitope or closely related sites on antigen 5, was observed. No such competition was detected with the sera from other helminthiasis. The sensitivity and specificity of CRIA was compared to that of ELISA and CRIA found to be an improved diagnostic test for hydatid disease.