Removing chemical and biological pollutants from swine wastewater through constructed wetlands aiming reclaimed water reuse.

Reusing reclaimed wastewater is needed to fight water scarcity, reduce freshwater consumption and conserve water resources, but one must ensure that hazardous substances are fully removed/eliminate before that reuse. The potential of lab-scale constructed wetlands (CWs) for the removal of chemical and biological contaminants from livestock wastewater, while maintaining nutrient levels for fertilization, was assessed, evaluating changes in microbial communities, with particular focus on potential pathogens. CW microcosms with two different substrates (lava rock or light expanded clay aggregate), both planted with Phragmites australis, were tested. After 15 days of treatment, removal rates were higher than 80% for Cd, Cr, Cu, Fe, Pb and Zn, in general with no significant differences between the two different substrates. Organic matter and nutrients were also removed but their levels still allowed the used of the treated wastewater as a fertilizer Removal of bacterial contamination was estimated through enumeration of cultivable bacteria. High removal rates of fecal indicator bacteria were observed, reaching >95% for enterococci and >98% for enterobacteria after 15 days of treatment, decreasing hazardous biological contaminants initially present in the wastewater. In addition, the microbial communities in the initial and treated wastewater, and in the plant roots bed substrate, were characterized by using 16SrRNA gene amplicon sequencing. Microbial communities in the CW systems showed a clear shift comparatively with the initial wastewater showing system adaptation and removal potentialities. This also revealed an important removal of the most represented potential pathogenic genus, Clostridium, which relative abundance decreased from 33% to 1% through the treatment. Overall, CWs showed potential to be efficient in removing chemical and biological contaminants, while maintaining moderated levels of nutrients, allowing the reuse of reclaimed water in agriculture, namely as fertilizer. Current results will contribute for the optimization and use of CWs for a sustainable treatment of liquid wastes, promoting the circular economy.

Tailoring of Mesoporous Silica-Based Materials for Enhanced Water Pollutants Removal.

The adsorptive performance of mesoporous silica-based materials towards inorganic (metal ions) and organic (dyes) water pollutants was investigated. Mesoporous silica materials with different particle size, surface area and pore volume were prepared and tailored with different functional groups. These materials were then characterised by solid-state techniques, namely vibrational spectroscopy, elemental analysis, scanning electron microscopy and nitrogen adsorption-desorption isotherms, allowing the successful preparation and structural modifications of the materials to be confirmed. The influence of the physicochemical properties of the adsorbents towards the removal of metal ions (Ni2+, Cu2+ and Fe3+) and organic dyes (methylene blue and methyl green) from aqueous solutions was also investigated. The results reveal that the exceptionally high surface area and suitable ζ-potential of the nanosized mesoporous silica nanoparticles (MSNPs) seem to favour the adsorptive capacity of the material for both types of water pollutants. Kinetic studies were performed for the adsorption of organic dyes by MSNPs and large-pore mesoporous silica (LPMS), suggesting that the process follows a pseudo-second-order model. The recyclability along consecutive adsorption cycles and the stability of the adsorbents after use were also investigated, showing that the material can be reused. Current results show the potentialities of novel silica-based material as a suitable adsorbent to remove pollutants from aquatic matrices with an applicability to reduce water pollution.

Atlas of the microbial degradation of fluorinated pesticides.

Fluorine-based agrochemicals have been benchmarked as the golden standard in pesticide development, prompting their widespread use in agriculture. As a result, fluorinated pesticides can now be found in the environment, entailing serious ecological implications due to their harmfulness and persistence. Microbial degradation might be an option to mitigate these impacts, though environmental microorganisms are not expected to easily cope with these fluoroaromatics due to their recalcitrance. Here, we provide an outlook on the microbial metabolism of fluorinated pesticides by analyzing the degradation pathways and biochemical processes involved, while also highlighting the central role of enzymatic defluorination in their productive metabolism. Finally, the potential contribution of these microbial processes for the dissipation of fluorinated pesticides from the environment is also discussed.

Advanced oxidation technologies and constructed wetlands in aquaculture farms: What do we know so far about micropollutant removal?

Aquaculture is the fastest growing animal food-producing sector. Water is the central resource for aquaculture, and it is essential that its quality be preserved. Micropollutants (MPs) can reach aquaculture through anthropogenic addition or inlet water, and may cause harmful effects such as endocrine disruption and antibiotic resistance, adversely affecting the fish species being farmed. Furthermore, the discharge of aquaculture effluents into the environment may contribute to the deterioration of water courses. In this sense, the implementation of environmentally responsible measures in aquaculture farms is imperative for the protection of ecosystems and human health. The European Commission (EC) has recently launched a guiding document promoting ecological aquaculture practices; however, options for water treatment are still lacking. Conventional processes are not designed to deal with MPs; this review article consolidates relevant information on the application of advanced oxidation technologies (AOTs) and constructed wetlands (CWs) as potential strategies in this regard. Although 161 studies on the application of AOTs or CWs in aquaculture have already been published, only 34 focused on MPs (28 on AOTs and 6 on CWs), whereas the others reported the removal of contaminants such as bacteria, organic matter, solids and inorganic ions. No study coupling both treatments has been reported to date for the removal of MPs from aquaculture waters. AOTs and CWs are prospective alternatives for the treatment of aquacultural aqueous matrices. However, the type of aquaculture activity and the specifications of these available technologies should be considered while selecting the most suitable treatment option.

Optimization of an Analytical Protocol for the Extraction of Microplastics from Seafood Samples with Different Levels of Fat.

Marine organisms are affected by the ubiquitous occurrence of microplastics (MPs) in the environment. Several protocols have been described to extract and quantify MPs in seafood, although their complex matrices, with high level of fat, can compromise the efficiency of MPs extraction. To solve this issue, the present study aimed to develop a detailed methodology suitable to process seafood samples with different levels of fat, namely fish and molluscs, from fresh and canned sources, including the immersive liquids from the cans. Sample digestion was tested using different solutions (10% KOH, 30% H2O2), temperatures (40 °C, 65 °C) and incubation times (24, 48, 72 h). For fat removal, three detergents (two laboratory surfactants and a commercial dish detergent) and 96% ethanol were tested, as well as the manual separation of fat. The methodology optimized in this study combined a digestion with 30% H2O2 at 65 °C, during 24 to 48 h, with a manual separation of the fat remaining after the digestion. All steps from the present methodology were tested in six types of polymers (PE-LD, PET, PE, AC, PS, and lycra), to investigate if these procedures altered the integrity of MPs. Results showed that the optimized methodology will allow for the efficient processing of complex seafood samples with different fat levels, without compromising MPs integrity (recoveries rate higher than 89% for all the polymers tested).

Severe Cutaneous Adverse Drug Reactions in Children: Epidemiological, Clinical and Etiological Aspects in Dermatology-Venereology Unit at National and Teaching Hospital of Cotonou.

BACKGROUND: The aim of this study was to describe the epidemiological, clinical and etiological aspects of severe cutaneous adverse drug reactions in children in dermatologyvenereology unit at National and Teaching Hospital of Cotonou. METHODS: A retrospective and descriptive study was carried out for 10 years in dermatology-venereology unit at the National and Teaching Hospital of Cotonou to document the epidemiological, clinical and etiological aspects of severe cutaneous adverse drug reactions in children. It included all children aged from 0 to 18 years with clinical diagnosis of severe cutaneous adverse drug reactions. Drug imputability was based on the criteria of the French pharmacovigilance group. RESULTS: Severe cutaneous adverse drug reactions accounted for 47.3% of paediatric cases (35/74 cases). The mean age was 9.3 years ± 5.2. The sex-ratio was 1.1. Self-medication was noted in 76.5% of children, on the initiative of parents in 66.7% of cases. There were 51.4% cases of Steven Johnson syndrome, 22.8% cases of Lyell syndrome, 8.5% cases of generalized and bullous fixed drug eruption, 2.9% cases of acute generalized exanthematous pustulosis and erythrodermic maculo-papular rash. Drug combinations was noted in 20% of cases. Penicillins (26.5%), paracetamol and sulfonamides (16.3%) were the drugs frequently incriminated. CONCLUSION: Steven Johnson syndrome and Lyell syndrome were the main severe cutaneous adverse drug reactions in children, mostly of school age. Penicillins, paracetamol and sulfonamides were the drugs frequently used and administered most often on self-medication.

INTRODUCTION: L'objectif de cette étude était de décrire les aspects épidémiologiques, cliniques et étiologiques des toxidermies graves chez les enfants en dermatologie à Cotonou. METHODES: Une étude rétrospective et descriptive a été réalisée sur 10 ans dans le service de dermatologie du Centre National Hospitalier et Universitaire de Cotonou pour documenter les aspects épidémiologiques, cliniques et étiologiques des toxidermies graves chez les enfants. Étaient inclus tous les enfants âgés de 0-18 ans chez qui le diagnostic clinique de toxidermie grave a été retenu. L'imputabilité médicamenteuse était basée sur les critères du groupe français de pharmacovigilance. RESULTATS: Les toxidermies graves représentaient 47,3% des cas pédiatriques (35/74 cas). L'âge moyen était de 9,3 ans ± 5,2. La sex-ratio H/F était de 1,1. Une automédication a été notée chez 76,5% des enfants, sur l'initiative des parents dans 66,7% des cas. Il y avait 51,4% de cas de syndrome de Steven Johnson, 22,8% de cas de syndrome de Lyell, 8,5% de cas d'érythème pigmenté fixe bulleux étendu, 2,9% de pustulose exanthématique aigüe généralisée et d'exanthème maculo-papuleux eìrythrodermique. Une polymédication a été notée dans 20% des cas. Les pénicillines (26,5%), le paracétamol et les sulfamides (16,3%) étaient les médicaments fréquemment incriminés. CONCLUSION: Le syndrome de Steven Johnson et le syndrome de Lyell étaient les principales toxidermies graves chez les enfants, majoritairement en âge scolaire. Les pénicillines, le paracétamol et les sulfamides étaient les médicaments fréquemment incriminés et administrés le plus souvent en automédication. Mots clés: Toxidermies graves, syndrome de Steven Johnson, syndrome de Lyell, automédication, sulfamides, enfants, Benin.

Pathogenic Microbial Profile and Antibiotic Resistance Associated with Periodontitis.

ABSTRACT: Phenotyping based on conventional microbiological, physiological, and molecular analysis by using ARDRA technique was developed with the aim to assess the pathogenic microbial load associated with different stages of the periodontal disease. In addition, in the face of the global issue of antimicrobial resistance, the isolated bacterial strains were evaluated for their antibiotic susceptibility profile. The pathogenic bacterial community was predominantly of Gram-negative strains (66.66%). The most common species were Citrobacter freundii, Bacillus sp., Raoutella sp., Klebsiella ozaenae and Pseudomonas sp. However, except for the healthy control group, Staphylococcus spp. was isolated from all stages of periodontitis. Multidrug resistance to beta-lactam antibiotics was observed for Streptococcus pneumoniae, Raoutella sp. and Enterococcus avium. Here, we verify a statistically significant relationship between periodontitis stages and the diversity of the bacterial community. Patients with periodontitis showed a more diverse and numerous bacterial community compared to healthy patients. In this sense, we reinforce that biofilms that harbour multidrug-resistant bacteria are a major concern in relation to restoring patient health. Thus, prophylactic measures for maintaining oral health are still the best option for reduce the risk of disease.

Beneficial Effects of Soluble Guanylyl Cyclase Stimulation and Activation in Sickle Cell Disease Are Amplified by Hydroxyurea: In Vitro and In Vivo Studies.

The complex pathophysiology of sickle cell anemia (SCA) involves intravascular hemolytic processes and recurrent vaso-occlusion, driven by chronic vascular inflammation, which result in the disease's severe clinical complications, including recurrent painful vaso-occlusive episodes. Hydroxyurea, the only drug frequently used for SCA therapy, is a cytostatic agent, although it appears to exert nitric oxide/soluble guanylyl cyclase (sGC) modulating activity. As new drugs that can complement or replace the use of hydroxyurea are sought to further reduce vaso-occlusive episode frequency in SCA, we investigated the effects of the sGC agonists BAY 60-2770 (sGC activator) and BAY 41-2272 (sGC stimulator) in the presence or absence of hydroxyurea on SCA vaso-occlusive mechanisms and cell recruitment both ex vivo and in vivo. These agents significantly reduced stimulated human SCA neutrophil adhesive properties ex vivo in association with the inhibition of surface ß2-integrin activation. A single administration of BAY 60-2770 or BAY 41-2272 decreased tumor necrosis factor cytokine-induced leukocyte recruitment in a mouse model of SCA vaso-occlusion. Importantly, the in vivo actions of both agonists were significantly potentiated by the coadministration of hydroxyurea. Erythroid cell fetal hemoglobin (HbF) elevation is also a major goal for SCA therapy. BAY 41-2272 but not BAY 60-2770 at the concentrations employed significantly induced γ-globin gene transcription in association with HbF production in cultured erythroleukemic cells. In conclusion, sGC agonist drugs could represent a promising approach as therapy for SCA, for use either as stand-alone treatments or in combination with hydroxyurea. SIGNIFICANCE STATEMENT: This preclinical study demonstrates that stimulators and activators of sGC are potent inhibitors of the adhesion and recruitment of leukocytes from humans and in mice with sickle cell anemia (SCA) and may represent a promising approach for diminishing vaso-occlusive episode frequency in SCA. Hydroxyurea, a drug already frequently used for treating SCA, was found to potentiate the beneficial effects of sGC agonists in in vivo studies, implying that these classes of compounds could be used alone or in combination therapy.

Biodegradation of enrofloxacin by microbial consortia obtained from rhizosediments of two estuarine plants.

This study aimed to investigate the potential of microbial communities from the rhizosediment of two plants - Phragmites australis and Juncus maritimus - occurring in an estuarine area subjected to a high anthropogenic impact, to biodegrade ENR, a commonly used veterinary antibiotic. An enrichment process with 1 mgL-1 of ENR was conducted during ca. 9 months, using acetate as a co-substrate. After this, the enriched microbial consortia were challenged with higher ENR concentrations of 2 and 3 mgL-1. Microbial cultures enriched with 1 mgL-1 of ENR were capable of biodegrading this antibiotic, though not completely. By the end of the enrichment phase, microbial cultures were defluorinating an average of 50% of the ENR supplemented. Higher ENR concentrations led to lower biodegradation performances, suggesting a possible toxic/inhibitory effect in the microbial cultures. Phylogenetic identification of the microorganisms isolated from microbial cultures enriched with ENR revealed a high taxonomical diversity, with microorganisms belonging mainly to Proteobacteria and Bacteroidetes phyla. Assemblage of the obtained isolated strains (according to the enriched cultures from which they were isolated) revealed that the resulting consortia were also capable of degrading ENR, indicating that the main microbial players in the biodegradation of this antibiotic were isolated. These consortia also showed to be more robust to degrade higher concentrations of ENR than the corresponding enriched cultures. This study shows that microorganisms derived from rhizosediments of the selected plants, exhibit capacity to biodegrade ENR, though not completely for the concentrations tested, and may be further explored for the development of bioremediation strategies for the treatment of this antibiotic.

Bioaccumulation of persistent and emerging pollutants in wild sea urchin Paracentrotus lividus.

Marine pollution has been increasing as a consequence of anthropogenic activities. The preservation of marine ecosystems, as well as the safety of harvested seafood, are nowadays a global concern. Here, we report for the first time the contamination levels of a large set of 99 emerging and persistent organic contaminants (butyltins (BTs), polycyclic aromatic hydrocarbons (PAHs), pesticides including pyrethroids, pharmaceuticals and personal care products (PCPs) and flame retardants) in roe/gonads of sea urchin Paracentrotus lividus. Sea urchins are a highly prized worldwide delicacy, and the harvesting of this seafood has increased over the last decades, particularly in South West Atlantic coast, where this organism is harvested mainly for exportation. Sampling was performed in three harvesting sites of the NW Portuguese coast subjected to distinct anthropogenic pressures: Carreço, Praia Norte and Vila Chã, with sea urchins being collected in the north and south areas of each site. Butyltins and pharmaceuticals were not found at measurable levels. Several PAHs, four pyrethroids insecticides, four PCPs and eleven flame retardants were found in roe/gonads of sea urchins, though in general at low levels. Differences among harvesting sites and between areas within each site were found, the lowest levels of contaminants being registered in Carreço. The accumulation of contaminants in sea urchins' roe/gonads seemed to reflect the low anthropogenic pressure felt in the sampling sites. Nevertheless, taking into account the low accumulated levels of chemicals, results indicate that sea urchins collected in South West Atlantic coast are safe for human consumption.

Macro and trace elements in Paracentrotus lividus gonads from South West Atlantic areas.

Sea urchin represents one of the most valuable seafood product being harvested and explored for their edible part, the gonads or roe. This species is generally considered a sentinel organism for ecotoxicological studies being widely used in monitoring programs to assess coastal aquatic environments quality, because is directly exposed to anthropogenic contaminants in their habitat. In this context, the aim of this study is to evaluate the concentrations of macro (Cl, K, P, Ca, S) and trace (Zn, Br, Fe, Sr, I, Se, Rb, Cu, Cr, Ni, As, iAs, Cd, Pb, Hg) elements in Paracentrotus lividus gonads from three South West Atlantic production areas subjected to distinct environmental and anthropogenic pressures. In all studied areas, the elements profile in sea urchin gonads was Cl > K > P > Ca > S > Zn > Br > Fe > Sr > I > Rb > Cu > Se > Cr > Ni, suggesting an element guide profile with special interest for sea urchin farming development. Concerning toxic elements, the profile was the following: As > Cd > Pb > Hg > iAs. The results evidenced higher levels of Pb and Hg in open areas. Distinct area characteristics and anthropogenic pressures of production areas evidence the importance of biomonitoring contaminants, particularly toxic elements. In general, the levels of these elements were below maximum levels in foodstuffs (MLs) which pose a minimal health risk to consumers.

Efficacy of antimicrobial agents incorporated in orthodontic bonding systems: a systematic review and meta-analysis.

AIMS: The purpose of this study was to evaluate the efficacy of orthodontic bonding systems containing different antimicrobial agents, as well as the influence of antimicrobial agent incorporation in the bonding properties of these materials. METHODS: Eight databases were searched: PubMed (Medline), Web of Science, Scopus, Lilacs, Ibecs, BBO, Scielo and Google Scholar. Any study that evaluated antimicrobial activity in experimental or commercial orthodontic bonding systems was included. DATA EXTRACTION: Data were tabulated independently and in duplicated by two authors on pre-designed data collection form. DATA SYNTHESIS: The global analysis was carried out using a random-effects model, and pooled-effect estimates were obtained by comparing the standardised mean difference of each antimicrobial orthodontic adhesive with the respective control group. A p-value < .05 was considered as statistically significant. RESULTS: Thirty-two studies were included in the qualitative analysis; of these, 22 studies were included in the meta-analysis. Antimicrobial agents such as silver nanoparticles, benzalkonium chloride, chlorhexidine, triclosan, cetylpyridinium chloride, Galla chinensis extract, acid ursolic, dimethylaminododecyl methacrylate, dimethylaminohexadecyl methacrylate, 2-methacryloyloxyethyl phosphorylcholine, 1,3,5-triacryloylhexahydro-1,3,5-triazine, zinc oxide and titanium oxide have been incorporated into orthodontic bonding systems. The antimicrobial agent incorporation in orthodontic bonding systems showed higher antimicrobial activity than the control group in agar diffusion (overall standardised mean difference: 3.71; 95% CI 2.98 to 4.43) and optical density tests (0.41; 95% CI -0.05 to 0.86) (p < .05). However, for biofilm, the materials did not present antimicrobial activity (6.78; 95% CI 4.78 to 8.77). Regarding bond strength, the global analysis showed antimicrobial orthodontic bonding systems were statistically similar to the control. CONCLUSIONS: Although there is evidence of antibacterial activity from in vitro studies, clinical and long-term studies are still necessary to confirm the effectiveness of antibacterial orthodontic bonding systems in preventing caries disease.

Mitochondrial DNA analysis of Rhipicephalus sanguineus s.l. from the western Iberian peninsula.

Rhipicephalus sanguineus Latreille (1806) (Ixodida: Ixodidae) is considered to be the most widely distributed tick and to have a vast range of habitats and hosts, including livestock, pets and wildlife. In addition to morphological differences, recent investigations using approaches based on molecular genetic markers have revealed the existence of different R. sanguineus lineages in different geographic regions. In this study, 475 ticks collected from dogs in the western Iberian peninsula were studied both morphologically and genetically, using 12S and 16S rDNA and COI gene markers in order to clarify the controversy over the systematic status of R. sanguineus sensu lato in Western Europe, and to compare the present data with those sourced from studies conducted in other regions of the world. Despite the high morphometric variability, particularly on spiracles in both genders and in female genitalia, data obtained with different genetic molecular markers show very low variability, suggesting the existence of a unique species. In addition, the phylogenetic analysis showed genetic uniformity, supporting the existence of a well-defined clade consisting of R. sanguineus s.l. specimens from Western Europe that are distinct from R. sanguineus s.l. from Africa. Furthermore, these data corroborate the existence of a polymorphic species in Western Europe, which requires to be consensually redescribed in view of its medical and veterinary importance in pathogen transmission.

Identification of stable fly attractant compounds in vinasse, a byproduct of sugarcane-ethanol distillation.

The stable fly, Stomoxys calcitrans (Diptera: Muscidae), is a worldwide pest of livestock. Recent outbreaks of stable flies in sugarcane fields in Brazil have become a serious problem for livestock producers. Larvae and pupae found inside sugarcane stems after harvesting may indicate that stable flies use these stems as potential oviposition or larval development sites. Field observations suggest that outbreaks of stable flies are associated with the vinasse and filter cake derived from biomass distillation in sugarcane ethanol production that are used as fertilizers in sugarcane fields. Adult stable flies are attracted to vinasse, which appears to present an ideal larval development site. The primary goal of the present study is to demonstrate the role of vinasse in influencing the sensory physiological and behavioural responses of stable flies, and to identify its associated volatile attractant compounds. Both laboratory and field studies showed that vinasse is extremely attractive to adult stable flies. Chemical analyses of volatiles collected revealed a wide range of carboxylic acids, alcohols, phenols and aldehydes as potential attractant compounds. These newly identified attractants could be used to develop a tool for the attractant-baited mass trapping of stable flies in order to reduce infestations.

Constructed wetlands for the removal of metals from livestock wastewater - Can the presence of veterinary antibiotics affect removals?

The presence of emergent antibiotics, in livestock wastewater may affect constructed wetlands (CWs) performance in the removal of other pollutants. The main objective of this study was to evaluate the influence of two antibiotics commonly used in livestock industry, enrofloxacin and ceftiofur, on metal removal by CWs. Microcosms (0.4m×0.3m×0.3m), simulating CWs, were constructed with Phragmites australis to treat livestock wastewater spiked or not with 100µg/L of enrofloxacin or ceftiofur (individually or in mixture). Wastewater was treated during 20 one-week cycles. After one-week cycle wastewater was removed and replaced by new wastewater (with or without spiking). At weeks 1, 2, 4, 8, 14, 18 and 20, treated wastewater was analysed to determine the removal rates of metals (Zn, Cu, Fe and Mn) and of each antibiotic. At weeks 1, 8 and 20 portions of the plant root substrate were collected and metals determined. At the end of the experiment metal levels were also determined in plant tissues. Removal rate of Fe from wastewater was 99%. Removal rates of Cu and Zn were higher than 85% and 89%, respectively, whereas for Mn removal rates up to 75% were obtained. In general, no significant differences were observed through time in the removals of the different metals, indicating that the systems maintained their functionality during the experimental period. Antibiotics did not interfere with the system depuration capacity, in terms of metals removals from wastewater, and ceftiofur even promoted metal uptake by P. australis. Therefore, CWs seem to be a valuable alternative to remove pollutants, including antibiotics and metals, from livestock wastewaters, reducing the risk the release of these wastewaters might pose into the environment, although more research should be conducted with other antibiotics in CWs.

Analysis of genetic diversity of Hyptis pectinata (L.) Poit. plants using ISSR markers.

Hyptis pectinata, popularly known as 'sambacaitá' or 'canudinho', is a medicinal and aromatic species widely used in the Brazilian Northeast. In Sergipe, the excessive extraction of natural resources may reduce the genetic variability of native plants. Thus, molecular markers have frequently been applied to the characterization of genetic diversity as the basis for germplasm conservation and breeding programs. The objective of the present study was to evaluate the genetic diversity of H. pectinata plants collected in different municipalities of the State of Sergipe using ISSR molecular markers. Thirty-four primers were tested, nine of which were selected for providing reproducible and analyzable amplification products, resulting in 67 polymorphic bands. The expected heterozygosity ranged from 0.32 to 0.45, with a mean of 0.39. Polymorphism information content was of 0.49, which classifies the markers as moderately informative. A dendrogram was constructed using unweighted pair group method with arithmetic mean, forming three clusters: Cluster I (79 plants); Cluster II (4 plants); and Cluster III (2 plants). Jaccard's similarity coefficients ranged from 0.06 to 0.98. The plants SAM-117 and SAM-119 presented greater similarity. Conversely, SAM-107 and SAM-171 were the most genetically distant. In general, H. pectinata plants collected in the State of Sergipe presented low to moderate genetic diversity.

Analysis of protein profile of tomato root infected with Fusarium oxysporum f. sp lycopersici.

Fusarium wilt caused by Fusarium oxysporum f. sp lycopersici (Fol) is one of the main diseases affecting tomatoes. The BHRS 2,3 genotype of tomato is, however, resistant to this disease. A proteomic approach was used to understand the defense mechanisms of this genotype using the tomato root, the first tissue that interacts with the fungus, as a target. Protein was extracted and separated by two-dimensional electrophoresis followed by staining with Coomassie brilliant blue. The proteins were identified by MALDI-TOF/TOF mass spectrometry. A total of 22 proteins were identified, 21 of which showed differential expression with 12 proteins being upregulated and nine being downregulated. Plants responded to the pathogen with increased expression of pathogenesis-related proteins. We noted the induction of proteins involved in hypersensitivity reaction and other defense mechanisms. The expression of proteins of primary metabolism related to energy production, however, decreased, as did the expression of two proteins related to defense against abiotic stress. These results demonstrate the presence of important mechanisms for defense against Fol in the tomato genotype BHRS 2,3.

Amphibian-killing chytrid in Brazil comprises both locally endemic and globally expanding populations.

Chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis (Bd), is the emerging infectious disease implicated in recent population declines and extinctions of amphibian species worldwide. Bd strains from regions of disease-associated amphibian decline to date have all belonged to a single, hypervirulent clonal genotype (Bd-GPL). However, earlier studies in the Atlantic Forest of southeastern Brazil detected a novel, putatively enzootic lineage (Bd-Brazil), and indicated hybridization between Bd-GPL and Bd-Brazil. Here, we characterize the spatial distribution and population history of these sympatric lineages in the Brazilian Atlantic Forest. To investigate the genetic structure of Bd in this region, we collected and genotyped Bd strains along a 2400-km transect of the Atlantic Forest. Bd-Brazil genotypes were restricted to a narrow geographic range in the southern Atlantic Forest, while Bd-GPL strains were widespread and largely geographically unstructured. Bd population genetics in this region support the hypothesis that the recently discovered Brazilian lineage is enzootic in the Atlantic Forest of Brazil and that Bd-GPL is a more recently expanded invasive. We collected additional hybrid isolates that demonstrate the recurrence of hybridization between panzootic and enzootic lineages, thereby confirming the existence of a hybrid zone in the Serra da Graciosa mountain range of Paraná State. Our field observations suggest that Bd-GPL may be more infective towards native Brazilian amphibians, and potentially more effective at dispersing across a fragmented landscape. We also provide further evidence of pathogen translocations mediated by the Brazilian ranaculture industry with implications for regulations and policies on global amphibian trade.

De novo transcriptome assembly for a non-model species, the blood-sucking bug Triatoma brasiliensis, a vector of Chagas disease.

High throughput sequencing (HTS) provides new research opportunities for work on non-model organisms, such as differential expression studies between populations exposed to different environmental conditions. However, such transcriptomic studies first require the production of a reference assembly. The choice of sampling procedure, sequencing strategy and assembly workflow is crucial. To develop a reliable reference transcriptome for Triatoma brasiliensis, the major Chagas disease vector in Northeastern Brazil, different de novo assembly protocols were generated using various datasets and software. Both 454 and Illumina sequencing technologies were applied on RNA extracted from antennae and mouthparts from single or pooled individuals. The 454 library yielded 278 Mb. Fifteen Illumina libraries were constructed and yielded nearly 360 million RNA-seq single reads and 46 million RNA-seq paired-end reads for nearly 45 Gb. For the 454 reads, we used three assemblers, Newbler, CAP3 and/or MIRA and for the Illumina reads, the Trinity assembler. Ten assembly workflows were compared using these programs separately or in combination. To compare the assemblies obtained, quantitative and qualitative criteria were used, including contig length, N50, contig number and the percentage of chimeric contigs. Completeness of the assemblies was estimated using the CEGMA pipeline. The best assembly (57,657 contigs, completeness of 80 %, <1 % chimeric contigs) was a hybrid assembly leading to recommend the use of (1) a single individual with large representation of biological tissues, (2) merging both long reads and short paired-end Illumina reads, (3) several assemblers in order to combine the specific advantages of each.