Evolution of complex symbiotic relationships in a morphologically derived family of lichen-forming fungi.

We studied the evolutionary history of the Parmeliaceae (Lecanoromycetes, Ascomycota), one of the largest families of lichen-forming fungi with complex and variable morphologies, also including several lichenicolous fungi. We assembled a six-locus data set including nuclear, mitochondrial and low-copy protein-coding genes from 293 operational taxonomic units (OTUs). The lichenicolous lifestyle originated independently three times in lichenized ancestors within Parmeliaceae, and a new generic name is introduced for one of these fungi. In all cases, the independent origins occurred c. 24 million yr ago. Further, we show that the Paleocene, Eocene and Oligocene were key periods when diversification of major lineages within Parmeliaceae occurred, with subsequent radiations occurring primarily during the Oligocene and Miocene. Our phylogenetic hypothesis supports the independent origin of lichenicolous fungi associated with climatic shifts at the Oligocene-Miocene boundary. Moreover, diversification bursts at different times may be crucial factors driving the diversification of Parmeliaceae. Additionally, our study provides novel insight into evolutionary relationships in this large and diverse family of lichen-forming ascomycetes.

Control of Phytophthora nicotianae with Mefenoxam, Fresh Brassica Tissues, and Brassica Pellets.

Sensitivity to mefenoxam, fresh Brassicaceae tissues, and Brassica pellets was evaluated in several isolates of Phytophthora nicotianae recovered from pepper and tomato plants. The isolates of P. nicotianae studied were classified as sensitive to mefenoxam, showing great variability among isolates in the 50 and 90% effective concentrations (EC50 and EC90, respectively). Sensitivity differentiated isolates from the two hosts of origin, being isolates from tomato plants more resistant to fungicide than those from pepper plants. This differentiation also occurred in the case of fresh Brassicaceae tissues assay. The most effective biofumigant in inhibiting mycelial growth of P. nicotianae isolates was Brassica nigra. The effectiveness of B. carinata, Sinapis alba, and B. oleracea varied depending on the dose. Isolates differed in susceptibility to compounds released by the Brassica pellets and then in the EC50 and EC90. No significant difference was found between the isolates depending on the host of origin. Greenhouse tests demonstrated the effectiveness of treatments with mefenoxam and with Brassica pellets to control P. nicotianae in pepper plants. Mefenoxam application could be a solution to the disease caused by P. nicotianae in tomato and pepper crops in this region but its use could increase resistance in populations. Biofumigation is a promising technique which can be further developed to form part of integrated pest management strategies.

Evaluation of the effectiveness of hospital discharge planning and follow-up in the primary care of patients with chronic obstructive pulmonary disease.

AIMS AND OBJECTIVES: To evaluate the effectiveness of protocolised intervention for hospital discharge and follow-up in the primary care of patients with chronic obstructive pulmonary disease (COPD). BACKGROUND: Chronic obstructive pulmonary disease is one of the main causes of morbidity and mortality internationally. DESIGN: A quasi-experimental design was adopted, with a control group and it was pseudorandomised by services. METHODS: Patients with COPD admitted to two tertiary-level public hospitals in Spain were recruited (2007-2008). The outcome variables included: readmission rate, patient satisfaction (LOPSS12), quality of life (St. George's Respiratory Questionnaire) and level of knowledge about COPD. 48 hours after admission, both groups were evaluated by specialist coordinating nurses. At the hospital, a coordinating nurse visited each patient in the experimental group every 24 hours to identify the main caregiver, provide information about the disease, explain treatment, identify care problems and needs and facilitate communication between professionals. 24 hours after discharge, the coordinating nurses informed the primary care nurses about patient discharge. The two nurses made the first home visit together. There were follow-up phone calls at 2, 6, 12 and 24 weeks after discharge. A total of 143 patients were recruited (Intervention group=56; Control group=87). RESULTS: The results showed a significant improvement in the evolution of quality of life, at 12 and 24 weeks after discharge; the level of knowledge about COPD revealed significant differences between the groups. There were no differences according to satisfaction or readmission rate. Multivariate analysis (non-conditional logistic regression) showed the intervention to be ineffective in reducing the readmission rate. CONCLUSIONS: The planning of discharge for patients with COPD is effective in terms of improving the patients' quality of life and level of knowledge about the disease. RELEVANCE TO CLINICAL PRACTICE: The characteristics of patients with COPD make it necessary to include them in hospital discharge planning programmes.

Multiple origins of high reciprocal symbiotic specificity at an intercontinental spatial scale among gelatinous lichens (Collemataceae, Lecanoromycetes).

Because the number of fungal species (mycobionts) exceeds the number of algae and cyanobacteria (photobionts) found in lichens by more than two orders of magnitude, reciprocal one-to-one specificity between one fungal species and one photobiont across their entire distribution is not expected in this symbiotic system, and has not previously been observed. The specificity of the cyanobacterium Nostoc found in lichens was evaluated at a broad geographical scale within one of the main families of lichen-forming fungi (Collemataceae) that associate exclusively with this photobiont. A phylogenetic study was conducted using rbcLXS sequences from Nostoc sampled from 79 thalli (representing 24 species within the Collemataceae), and 163 Nostoc sequences gathered from GenBank. Although most of the lichen-forming fungal species belonging to the Collemataceae exhibited the expected generalist pattern of association with multiple distinct lineages of Nostoc, five independent cases of one-to-one reciprocal specificity at the species level, including two that span intercontinental distributions, were discovered. Each of the five distinct monophyletic Nostoc groups, associated with these five highly specific mycobiont species, represent independent transitions from a generalist state during the evolution of both partners, which might be explained by transitions to asexual fungal reproduction, involving vertical photobiont transmission, and narrowing of ecological niches.

Genetic distances within and among species in monophyletic lineages of Parmeliaceae (Ascomycota) as a tool for taxon delimitation.

The species delimitation in fungi is currently in flux. A growing body of evidence shows that the morphology-based species circumscription underestimates the number of existing species. The large and ever growing number of DNA sequence data of fungi makes it possible to use these to identify potential cases of hidden species, which then need to be studied with extensive taxon samplings. We used Parmeliaceae, one of the largest families of lichenized fungi as a model. Intra- and interspecific distances derived from maximum-likelihood phylogenetic trees inferred from 491 nuclear ITS rDNA sequences were examined for five major clades of parmelioid lichens. The intra- and interspecific distances were well separated in most cases allowing the calculation of a threshold, with exceptions of highly deviating distances in a few cases. These situations are shown to be taxa in which the current delimitation needs revision. Thus the analysis of the distance distributions is shown to be a powerful tool for identifying species complexes.

Disentangling the Collema-Leptogium complex through a molecular phylogenetic study of the Collemataceae (Peltigerales, lichen-forming Ascomycota).

Family Collemataceae (Peltigerales, Ascomycota) includes species of cyanolichens with foliose to fruticose or crustose thalli, with simple or septate ascospores. The current classification divides this family into two groups on the basis of ascospore types. The objective of this study was to evaluate the phylogenetic relationships within this family. Combined DNA sequence data from the nuclear large subunit and mitochondrial small subunit ribosomal RNA genes were used to evaluate monophyly of the family and the relationships between the largest genera of this family. The results revealed that this family is not monophyletic. Genera Staurolemma and Physma, currently classified within the Collemataceae, were found nested within the Pannariaceae. The second result of this study confirms that the genera Collema and Leptogium, both part of the Collemataceae s. str., are not monophyletic and that the presence of a thallus cortex is not a synapomorphy for Leptogium. The main taxonomic conclusion is that families Collemataceae and Pannariaceae were recircumscribed in light of molecular findings with the latter family now including Staurolemma and Physma. Genera Collema and Leptogium form a single mixed monophyletic group. Inferred ancestral character states within the Collema-Leptogium complex revealed that the ancestor of this family had a thallus without cortex and that a cortex evolved at least twice relatively early in the evolution of the Collemataceae s. str. These independent gains of a thallus cortex seems to be associated with a transition from colonizing bare rocks and soils in semi-arid and exposed habitats to epiphytism in shady humid forests.

Isolation and genetic identification of PAH degrading bacteria from a microbial consortium.

Polycyclic aromatic hydrocarbons (PAH; naphthalene, anthracene and phenanthrene) degrading microbial consortium C2PL05 was obtained from a sandy soil chronically exposed to petroleum products, collected from a petrochemical complex in Puertollano (Ciudad Real, Spain). The consortium C2PL05 was highly efficient degrading completely naphthalene, phenanthrene and anthracene in around 18 days of cultivation. The toxicity (Microtox method) generated by the PAH and by the intermediate metabolites was reduced to levels close to non-toxic in almost 40 days of cultivation. The identified bacteria from the contaminated soil belonged to gamma-proteobacteria and could be include in Enterobacter and Pseudomonas genus. DGGE analysis revealed uncultured Stenotrophomonas ribotypes as a possible PAH degrader in the microbial consortium. The present work shows the potential use of these microorganisms and the total consortium for the bioremediation of PAH polluted areas since the biodegradation of these chemicals takes place along with a significant decrease in toxicity.

Panmixia and dispersal from the Mediterranean Basin to Macaronesian Islands of a macrolichen species.

The Mediterranean region, comprising the Mediterranean Basin and the Macaronesian Islands, represents a center of diversification for many organisms. The genetic structure and connectivity of mainland and island microbial populations has been poorly explored, in particular in the case of symbiotic fungi. Here we investigated genetic diversity and spatial structure of the obligate outcrossing lichen-forming fungus Parmelina carporrhizans in the Mediterranean region. Using eight microsatellite and mating-type markers we showed that fungal populations are highly diverse but lack spatial structure. This is likely due to high connectivity and long distance dispersal of fungal spores. Consistent with low levels of linkage disequilibrium and lack of clonality, we detected both mating-type idiomorphs in all populations. Furthermore we showed that the Macaronesian Islands are the result of colonization from the Mediterranean Basin. The unidirectional gene flow, though, seemed not to be sufficient to counterbalance the effects of drift, resulting in comparatively allelic poor peripheral populations. Our study is the first to shed light on the high connectivity and lack of population structure in natural populations of a strictly sexual lichen fungus. Our data further support the view of the Macaronesian Islands as the end of the colonization road for this symbiotic ascomycete.

Effect of biofumigation with brassica pellets combined with Brassicaceae cover crops and plastic cover on the survival and infectivity of inoculum of Phytophthora nicotianae Breda de Haan.

BACKGROUND: Biofumigation with defatted seed meal of Brassicaceae in the form of pellets has several advantages over the incorporation of fresh Brassicaceae crops to control soil-borne diseases. Two field experiments were established to evaluate the effect of biofumigation with brassica pellets on the survival and infectivity of Phytophthora nicotianae Breda de Haan inoculum introduced before treatments. In the spring experiment the incorporation of additional Brassicaceae cover crop (Brassica nigra L. and Sinapis alba L.) was tested, and in the summer experiment two brassica pellet doses were applied. RESULTS: Biofumigation with brassica pellets in spring (3000 kg ha(-1) with and without plastic) or in summer (3000 kg ha(-1) with or without plastic; 6000 kg ha(-1) without plastic) had no significant effect on the survival of P. nicotianae, regardless of the incorporation of additional Brassicaceae cover crop in spring. Reduction in infectivity in spring was related to the application of plastic, especially when combined with brassica pellets and Brassicaceae crop. In summer, soil temperature was the main factor in the inactivation of the inoculum, especially when plastic was applied, and no additional inactivation was achieved with brassica pellets. CONCLUSION: In spring and summer, biofumigation with brassica pellets had no effect on the survival of P. nicotianae. Application of plastic in spring may reduce infectivity. Soil temperature is the main factor in the inactivation of inoculum in summer, especially when plastic is applied. © 2015 Society of Chemical Industry.

Prevalence, awareness, treatment, and control of high low-density lipoprotein cholesterol in Brazil: Baseline of the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil).

BACKGROUND AND OBJECTIVES: Dyslipidemia is a pivotal risk factor for coronary heart disease (CHD). The purpose of this study was to identify the profile of dyslipidemia in a Brazilian population, according to high low-density lipoprotein (LDL-C) levels. We used the classification of the 2004 update of National Cholesterol Education Program Adult Treatment Panel III (ATP-III). METHODS: Of the 15,105 men and women aged 35 to 74 years enrolled in the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil), we included 14,648 subjects (97%). They had data to categorize them according to the NCEP-ATP-III criteria. We compared 4 categories: "0-1" risk factors, "2 or more risk factors", "CHD or CHD risk equivalent", and "CHD at very high risk". The sociodemographic determinants used were sex, age, ethnicity, income, education, and health insurance. Poisson regression was used to estimate the prevalence ratios for cholesterol (LDL-C), frequency, awareness, treatment, and control of high LDL-C. RESULTS: The frequencies of high LDL-C, awareness, treatment, and control were 45.5%, 58.1%, 42.3%, and 58.3%, respectively. After adjustment for sociodemographic determinants, the prevalence ratios for high LDL-C were significantly higher for men, blacks, older subjects, and subjects with lower levels of education. Low frequency of awareness, treatment, and uncontrolled values of LDL-C was observed among men, mixed race and blacks, poorer, less educated, and those who did not have private health insurance. CONCLUSIONS: The prevalence of high LDL-C was elevated in this Brazilian population, with low rates of awareness, treatment and control, and remarkable socioeconomic disparity.

Genetic variation in the widespread lichenicolous fungus Marchandiomyces corallinus.

The lichenicolous basidiomycete Marchandiomyces corallinus is widely distributed in North America and Europe, where it commonly is found on a variety of lichens. Theoretically either of these characteristics, a wide geographic range or generalized host ecology, could provide opportunities for genetic differentiation within this species. To determine how genetic variation is partitioned in M. corallinus, 12 fungal isolates were obtained from locations in North America and Europe; at two locations, in Washington County, Maine, and on the Isle of Mull in Scotland, fungi also were isolated from different lichen hosts. Vegetative mycelial compatibility tests were used to determine compatibility groupings from among the isolates; in addition, several PCR amplification products (RAPD, nuITS rDNA) were obtained for each isolate. A number of distinct compatibility groups were recognizable based on geography, not host ecology. In addition compatible isolates always were restricted to either North America or Europe. However RAPD markers indicated that compatible isolates are not always genetically identical. The presence of sequence heterozygosity at specific positions indicated that the isolates are heterokaryotic and a number of distinct haplotypes could be identified based on ITS variation at three separate locations. This type of genetic variation in these fungi suggests that sexual recombination is possible and that genetic differentiation has taken place recently as a result of geographic isolation, not host switching.

[Maternal autoimmune thyroid disease: relevance for the newborn]. / Enfermedad tiroidea autoinmunitaria materna: repercusión en el recién nacido.

BACKGROUND AND OBJECTIVE: Autoimmune thyroid disease is amongst the most frequent endocrine disorders during pregnancy. It is associated with an increase in perinatal morbidity, congenital defects, neurological damage, fetal and neonatal thyroid dysfunction. Maternal thyroid hormones play a key role in child neurodevelopment. We aimed to evaluate the thyroid function and the clinical course of neonates born from mothers with autoimmune thyroid disease during the first months of life in order to define the follow-up. PATIENTS AND METHOD: We monitored thyroid function and clinical status during the first months in 81 newborns of mothers with autoimmune thyroid disease; 16 had Graves disease and 65 autoimmune thyroiditis. RESULTS: A percentage of 4.93 newborns had congenital defects, and 8.64% neonates showed an increase in thyrotropin (TSH) (>9.5 µUI/mL 2 times) and required thyroxin within the first month of life. A 85.7% of these showed a negative newborn screening (due to a later increase of TSH). A higher TSH value in the newborn was related to an older age of the mother, higher levels of thyroid peroxidase (TPO) antibody during pregnancy and lower birth weight. A higher free thyroxine (FT4) value in the newborn was related to fewer days of life and mothers with Graves disease. CONCLUSIONS: We recommend the evaluation of TSH, T4 and TPO antibodies before 10 weeks in all pregnant women with follow-up if maternal thyroid autoimmunity or disorders is detected. It is also recommended to test children's serum TSH and FT4 at 48 h of life in newborns of mothers with autoimmune thyroid disease and repeat them between the 2nd and 4th week in children with TSH>6 µUI/mL. Careful endocrine follow-up is advised in pregnant women and children if hyperthyroidism is detected.

Non-developing ascospores in apothecia of asexually reproducing lichen-forming fungi.

The presence of apothecia in mixed species (vegetatively reproducing lichens, occasionally producing ascomata) has been interpreted as a mechanism to increase genetic variability in mostly clonal populations. However, spore viability from these apothecia has not been studied. We asked whether ascospores of the mixed species Physconia grisea are viable and thereby contribute to increasing the genetic diversity within populations of this species. An ontogenetic study of spores in cultures of P. grisea and a related sexual species (P. distorta), showed that although mature apothecia from both species produced and discharged meiospores capable of germination, spores from P. grisea were only rarely (0.43%) able to continue development whereas those from P. distorta germinated and developed successfully. The strongly reduced viability of P. grisea spores suggested that they do not have a strong reproductive function, at least in the two local populations analyzed. Additionally, we show that the segregation of Physconia grisea ssp. lilacina does not have molecular support.

Spigelia genuflexa (Loganiaceae), a new geocarpic species from the Atlantic forest of northeastern Bahia, Brazil.

A new species of Spigelia L. (Loganiaceae), Spigelia genuflexa Popovkin & Struwe, sp. n., from the Atlantic forest of northeastern Bahia, Brazil, is described, being the first reported geocarpic species in the family. During fruit maturation, the basal infructescences bend down towards the ground, depositing the fruit on the surface (and burying it in soft kinds of ground cover, e.g., moss), whereas the upper ones do so slightly but noticeably. The species is a short-lived annual apparently restricted to sandy-soil habitat of the Atlantic forest of northeastern Bahia, with variable and heterogeneous microenvironment and is known from only two restricted localities. A short review of amphi- and geocarpic species is provided. A discussion of comparative morphology within Spigelia with regards to dwarfism, indumentum, and annual habit is included. A phylogenetic parsimony and Bayesian analysis of ITS sequences from 15 Spigelia species plus 17 outgroups in Loganiaceae confirms its independent taxonomic status: on the basis of sequence similarity and phylogenetic topology it is phylogenetically distinct from all Spigelia species sequenced so far.

Phylogeography and divergence date estimates of a lichen species complex with a disjunct distribution pattern.

Disjunct species distributions may result from a combination of geologic events and long-distance dispersal. The foliose lichen species complex Leptogium furfuraceum-L. pseudofurfuraceum has an intercontinental disjunction pattern. Populations of this species complex are found in western North America, southern South America, Africa, and southern Europe. We conducted a phylogenetic study to reconstruct the biogeographic history of this species complex using two ribosomal genes (ITS and LSU) and a protein-coding gene (partial RPB2). Results indicated that the complex comprises four geographically restricted genetic lineages. A sister relationship was found between populations from the same hemispheres, incongruent with previous data derived from morphological characteristics and geographical classification schemes. Incorporating Bayesian ancestral area reconstruction and Bayesian divergence time estimation, we proposed an evolutionary hypothesis for the species complex. The results suggested that processes of biotic expansion via transoceanic dispersal were responsible for the species divergence and distribution patterns observed today. This study also expands the view that cryptic speciation is not a rare phenomenon among fungi and lichens.

Personality profiles and minor affective psychopathology in a non-clinical sample: an empirical verification of Cloninger's theoretical model.

BACKGROUND: Psychopathological vulnerability may be related to certain personality traits. The aim of this study was to explore the association of minor affective psychopathology and the regular use of psychotropic medication with temperament and character profiles from Cloninger's personality model, in a sample of active professional people. METHODS: This cross-sectional study included 498 non-clinical subjects, teachers in a local school system. Instruments used included the self-administered General Health Questionnaire (GHQ-28) to measure psychiatric morbidity; the Center for Epidemiologic Studies Depression scale (CES-D) to measure depressive symptoms; documentation of regular use of psychotropic medication; and the Temperament and Character Inventory (TCI-125) for personality traits self-assessment. RESULTS: The proportion of subjects presenting psychiatric morbidity (GHQ-28>6) or depressive symptoms (CES-D>20) was significantly higher among explosive, passive-aggressive, and obsessional temperament profiles, and among schizotypal, moody, melancholic and dependent character profiles. Similar results were observed with the scores on each of the four GHQ-28 subscales (depression, anxiety, social dysfunction, and somatic symptoms). The regular use of psychotropic medications was significantly higher among the passive-aggressive and explosive temperament types, and among the schizoptypal and moody character types. LIMITATIONS: Being a cross-sectional study, no causal attributions can be inferred. Subjects on sick leave were excluded, so the sample was not representative of the general population. The data were collected using self-reporting questionnaires, and no specific psychiatric diagnoses were obtained. CONCLUSIONS: It is possible to identify certain personality configurations associated with minor psychopathology and concomitant use of psychotropics, among active professional people.

Phylogenetic diversity of lichen-associated homobasidiomycetes.

The vast majority of lichenicolous fungi are relatively host-specific, nonvirulent ascomycetes and heterobasidiomycetes. A few known lichenicolous homobasidiomycetes (mushroom-forming fungi) generally exhibit broad host ecologies and in some cases, high virulence. Many produce conspicuous sclerotia or bulbils, thought to be adaptive in dispersal and survival. To more clearly understand the evolution of these atypical behaviors in lichenicolous basidiomycetes, we isolated or acquired specimens or cultures of 23 lichenicolous homobasidiomycetes and their relatives, from which we obtained mainly nuclear and some mitochondrial rDNA sequences. Phylogenetic analyses in this study indicate that a lichenicolous habit arose in four major clades. In two of these clades the habit represents a major evolutionary theme linked to the origin of well-known basidiolichens. The phylogenetic diversity of these fungi indicates that the lichenicolous habit arose recently and independently in the mushroom-forming fungi.