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.

The intensity and correlates of the feelings of loneliness in people with psychosis.

OBJECTIVE: Loneliness is an established risk factor for numerous negative health outcomes. The aims of the present study were to compare the levels of loneliness between patients with psychotic disorders and members of the general population and to identify factors independently associated with loneliness in psychosis. METHODS: A total of 207 patients with psychotic disorders recruited between February 2013 and February 2015 from inpatient and day wards and an outpatient clinic of the Institute of Psychiatry and Neurology (IPN) in Warsaw (Poland) were included in this cross-sectional study. They were administered the De Jong Gierveld Loneliness Scale (DJGLS) and a set of instruments assessing three types of explanatory variables: socio-demographic, psychosocial and psychiatric (clinical). The comparison group was a random sample of 20,000 inhabitants of Poland who took part in the Generations and Gender Survey (GGS-PL) conducted in 2011. The two groups were matched for socio-demographic characteristics. RESULTS: The patient sample proved to be significantly lonelier than the general population sample. The higher level of loneliness in people with psychotic disorders was most strongly related to psychosocial factors, particularly more severe internalized stigma and lesser social support, followed by worse interpersonal competence and smaller social network. After adjusting for socio-demographic and psychosocial variables, the only clinical variable significantly associated with more intense feelings of loneliness turned out to be the greater number of psychiatric inpatient admissions. The findings did not lend support to the view that socio-demographics play a major role in explaining variation in loneliness in psychosis. CONCLUSIONS: People with psychotic disorders are predisposed to experiencing elevated levels of loneliness. To counteract the pernicious effects of this on their health and well-being, there is a need for comprehensive therapeutic programs targeting self-stigma, enhancing social support networks and improving social skills.

Bridgehead invasion of a monomorphic plant pathogenic bacterium: Xanthomonas citri pv. citri, an emerging citrus pathogen in Mali and Burkina Faso.

Molecular epidemiology studies further our understanding of migrations of phytopathogenic bacteria, the major determining factor in their emergence. Asiatic citrus canker, caused by Xanthomonas citri pv. citri, was recently reported in Mali and Burkina Faso, a region remote from other contaminated areas. To identify the origin and pathways of these emergences, we used two sets of markers, minisatellites and microsatellites, for investigating different evolutionary scales. Minisatellite typing suggested the introduction of two groups of strains in Mali (DAPC 1 and DAPC 2), consistent with microsatellite typing. DAPC 2 was restricted to Bamako district, whereas DAPC 1 strains were found much more invasive. The latter strains formed a major clonal complex based on microsatellite data with the primary and secondary founders detected in commercial citrus nurseries and orchards. This suggests that human activities played a major role in the spread of DAPC 1 strains via the movement of contaminated propagative material, further supported by the frequent lack of differentiation between populations from geographically distant nurseries and orchards. Approximate Bayesian Computation analyses supported the hypothesis that strains from Burkina Faso resulted from a bridgehead invasion from Mali. Multi-locus variable number of tandem repeat analysis and Approximate Bayesian Computation are useful for understanding invasion routes and pathways of monomorphic bacterial pathogens.

Experiences of discrimination and the feelings of loneliness in people with psychotic disorders: the mediating effects of self-esteem and support seeking.

OBJECTIVE: People with psychotic disorders frequently become targets of discrimination, which may have devastating effects on their social relations and lead to the feelings of loneliness. This study has explored whether self-esteem and support seeking serve as mediators in the relationship between experiences of discrimination and loneliness. METHODS: A total of 110 persons with psychotic disorders (International Classification of Diseases, 10th Revision (ICD-10): F20-F29) were evaluated with self-report measures of discrimination experiences, self-esteem, support seeking and loneliness. The relationships between variables were examined with path modeling. Bootstrap mediation analyses were used for testing the statistical significance of indirect effects. RESULTS: Experiences of discrimination have been demonstrated to increase the level of loneliness both directly and indirectly. The indirect effect of discrimination on loneliness via self-esteem decrement has been proven to be significant. Support has been also found for a mediation model in which discrimination experiences negatively affect self-esteem, an undermined self-esteem diminishes the tendency to seek social support, and reduced support seeking worsens the sense of loneliness. However, discrimination experiences and support seeking have turned out to be unrelated and thus the hypothesized indirect effect of discrimination on loneliness through the weakening of the willingness to seek social support has not been confirmed by the data. CONCLUSIONS: The findings contribute to the understanding of the mechanisms through which experiences of discrimination aggravate perceived social isolation. Self-esteem and the tendency to seek social support have emerged as possible targets for interventions aiming to counteract the negative influence of rejection experiences on social relationships of people with psychotic disorders.

New multilocus variable-number tandem-repeat analysis tool for surveillance and local epidemiology of bacterial leaf blight and bacterial leaf streak of rice caused by Xanthomonas oryzae.

Multilocus variable-number tandem-repeat analysis (MLVA) is efficient for routine typing and for investigating the genetic structures of natural microbial populations. Two distinct pathovars of Xanthomonas oryzae can cause significant crop losses in tropical and temperate rice-growing countries. Bacterial leaf streak is caused by X. oryzae pv. oryzicola, and bacterial leaf blight is caused by X. oryzae pv. oryzae. For the latter, two genetic lineages have been described in the literature. We developed a universal MLVA typing tool both for the identification of the three X. oryzae genetic lineages and for epidemiological analyses. Sixteen candidate variable-number tandem-repeat (VNTR) loci were selected according to their presence and polymorphism in 10 draft or complete genome sequences of the three X. oryzae lineages and by VNTR sequencing of a subset of loci of interest in 20 strains per lineage. The MLVA-16 scheme was then applied to 338 strains of X. oryzae representing different pathovars and geographical locations. Linkage disequilibrium between MLVA loci was calculated by index association on different scales, and the 16 loci showed linear Mantel correlation with MLSA data on 56 X. oryzae strains, suggesting that they provide a good phylogenetic signal. Furthermore, analyses of sets of strains for different lineages indicated the possibility of using the scheme for deeper epidemiological investigation on small spatial scales.

First Report of Xanthomonas citri pv. mangiferaeindicae Causing Mango Bacterial Canker on Mangifera indica in Ivory Coast.

Xanthomonas citri pv. mangiferaeindicae causing bacterial canker (or black spot) is a major mango (Mangifera indica L.) pathogen in tropical and subtropical areas (3). The bacterium infects a wide range of mango cultivars, and induces raised, angular, black leaf lesions, sometimes with a yellow chlorotic halo. Fruit symptoms first appear as small water-soaked spots on the lenticels turning into star-shaped, erumpent lesions, which exude an infectious gum, yielding tear-stain patterns. Severe infections cause severe defoliation and/or premature fruit drop. Twig cankers are potential sources of inoculum and weaken branch resistance to winds. Drastic yield losses have been reported at grove scale for susceptible cultivars (3). Mango leaves showing typical angular, black, raised leaf lesions were first observed and collected in April 2014 from trees cv. Kent in five localities of the Korhogo province of Ivory Coast (i.e., the major commercial mango-growing area in this country). Non-pigmented Xanthomonas-like colonies were isolated on KC semi-selective medium (4). Five strains (LL60-1, LL61-1, LL62-1, LL63-1, and LL64-1), one from each locality, were compared by multilocus sequence analysis (MLSA) to the type strain of X. citri and the pathotype strain of several X. citri pathovars, including pvs. anacardii and mangiferaeindicae. This assay targeted the atpD, dnaK, efp, and gyrB genes, as described previously (2). Nucleotide sequences were 100% identical to those of the pathotype strain of X. citri pv. mangiferaeindicae whatever the gene assayed, but differed from any other assayed X. citri pathovar. Leaves of mango cv. Maison Rouge from the youngest vegetative flush were infiltrated (10 inoculation sites/leaf for three replicate leaves on different plants/bacterial strain) as detailed previously (1) with the same five strains. Bacterial suspensions (~1 × 105 cfu/ml) were prepared in 10 mM Tris buffer (pH 7.2) from 16-h-old cultures on YPGA (7 g yeast, 7 g peptone, 7 g glucose, and 18 g agar/liter, pH 7.2). The negative control treatment consisted of three leaves infiltrated with sterile Tris buffer (10 sites/leaf). Plants were incubated in a growth chamber at 30 ± 1°C by day and 26 ± 1°C by night (12-h day/night cycle) at 80 ± 5% RH. All leaves inoculated with the strains from Ivory Coast showed typical symptoms of bacterial canker a week after inoculation. No lesions were recorded from the negative controls. The pathogen was recovered at high population densities (>1 × 106 cfu/lesion) from leaf lesions, typical of a compatible interaction (1) and isolated colonies were identified as the target by atpD sequencing (2). Koch's postulates have therefore been fully verified. This is the first report of the disease in Ivory Coast, a country which has been an internationally significant mango exporter (up to 15,000 tons per year) over the last two decades. A high disease incidence and severity were observed, outlining the need for implementing integrated pest management in mango groves and the production of disease-free nursery stock. This report further expands the distribution of the pathogen in West Africa after its first description from Ghana in 2011 (5) and subsequently in other neighboring countries. References: (1) N. Ah-You et al. Phytopathology 97:1568, 2007. (2) L. Bui Thi Ngoc et al. Int. J. Syst. Evol. Microbiol. 60:515, 2010. (3) L. Gagnevin and O. Pruvost. Plant Dis. 85:928, 2001. (4) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005. (5) O. Pruvost et al. Plant Dis. 95:774, 2011.

First Report of Xanthomonas citri pv. citri Pathotype A Causing Asiatic Citrus Canker in Grande Comore and Anjouan.

The causal agent of Asiatic citrus canker, Xanthomonas citri pv. citri, is a bacterium of major economic importance in tropical and subtropical citrus-producing areas. X. citri pv. citri pathotype A can cause severe infection in a wide range of citrus species and induces erumpent, callus-like lesions with water-soaked margins evolving to corky cankers and leading to premature fruit, leaf drop, and twig dieback on susceptible cultivars. This quarantine organism can strongly impact citrus markets so it has consequently been subjected to eradication efforts and international quarantine regulations. Asiatic citrus canker occurs on most islands in the Southwest Indian Ocean region including the Mascarene and Seychelles archipelagos. In the Comoros archipelago, the disease was observed for the first time in Mohéli island in 1966 (2), but had not yet been reported in neighboring islands, Grande Comore and Anjouan. In September 2013, leaves of key lime (Citrus aurantifolia) and sweet orange (C. sinensis) showing symptoms of citrus canker were collected from Anjouan, Grande Comore, and Mohéli. Nine Xanthomonas-like strains (three from each of the three islands) were isolated using KC semi-selective medium (5) from diseased samples (LK126-3, LK127-7, LK128-2, LK131-10, LK137-1, LK141-3, LK144-5, LK145-5, LK146-2). Based on a specific PCR assay with 4/7 primers (4), all Xanthomonas-like strains were tentatively identified as X. citri pv. citri. All strains produced a 468-bp amplicon similar to X. citri pv. citri strain IAPAR 306 used as a positive control. Negative control reactions with sterile tris buffer did not produce amplicons. Multilocus sequence analysis (MLSA) targeting six housekeeping genes (atpD, dnaK, efp, gltA, gyrB, and lepA) (1,3) fully identified all strains from the Comoros as X. citri pv. citri. More specifically, eight strains were identified as sequence type ST2 composed of pathotype A strains of X. citri pv. citri (3) (including all strains from the Southwest Indian Ocean region) while one of them (LK141-3 from Mohéli) was identified as a new sequence type based on a non-synonymous single nucleotide polymorphism in gyrB (accession KJ941208). All strains were inoculated by a detached leaf assay (3) onto Mexican lime SRA 140 (C. aurantifolia), Tahiti lime SRA 58 (C. latifolia), sweet orange New Hall Navel SRA 343 (C. sinensis), grapefruit Henderson SRA 336 (C. paradisi), and Ortanique tangor SRA 110 (C. reticulata × C. sinensis). All citrus species inoculated produced typical erumpent, callus-like tissue at wound sites. Xanthomonas-like yellow colonies were re-isolated from lesions produced on Mexican lime. Boiled bacterial suspensions were assayed by PCR with 4/7 primers (4) and produced the expected amplicon, fulfilling Koch's postulates. No lesions developed on the negative control consisting of inoculations with sterile tris buffer. This is the first report of X. citri pv. citri-A causing Asiatic citrus canker in Grande Comore and Anjouan islands confirming the wide distribution of the pathogen in Southwest Indian Ocean islands. Canker-free nurseries and grove sanitation should be implemented to decrease the prevalence of Asiatic canker in the Comoros. References: (1) N. F. Almeida et al. Phytopathology 100:208, 2010. (2) J. Brun. Fruits 26:533, 1971. (3) L. Bui Thi Ngoc et al. Int. J. Syst. Evol. Microbiol. 60:515, 2010. (4) J. S. Hartung et al. Phytopathology 86:95, 1996. (5) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005.

Extent and predictors of stigma experienced by patients with schizophrenia.

BACKGROUND: Schizophrenia is regarded as one of the most stigmatized mental illnesses. Relatively few studies have investigated actual stigma experiences among people with schizophrenia and the factors which may contribute to it. AIMS: This cross-sectional study assesses the extent of stigma experienced by patients with schizophrenia and attempts to establish its clinical and sociodemographic predictors. METHODS: A total of 153 subjects with schizophrenia (62 outpatients and 91 inpatients) were evaluated with the use of the Stigma section of the Consumer Experiences of Stigma Questionnaire (CESQ) and several instruments measuring their subjective quality of life, social functioning and severity of psychiatric symptoms. RESULTS: Stigmatization experiences were common among respondents who most frequently reported having concealed their illness (86%), witnessed others saying offensive things about the mentally ill (69%), worried about being viewed unfavorably (63%) and been treated as less competent (59%). Higher levels of stigma were related to lower subjective quality of life and younger age of illness onset. No significant associations were found between stigma and symptoms or level of social functioning. CONCLUSION: Our findings point at the reduced life satisfaction as a key aspect of the subjective experience of the stigma of schizophrenia.