The damage caused by Cd toxicity to photosynthesis, cellular ultrastructure, antioxidant metabolism, and gene expression in young cacao plants are mitigated by high Mn doses in soil.

Manganese (Mn) is one of the essential mineral micronutrients most demanded by cacao. Cadmium (Cd) is highly toxic to plants and other living beings. There are indications that Mn can interact with Cd and mitigate its toxicity. The objective of this study was to evaluate the action of Mn on the toxic effect of Cd in young plants of the CCN 51 cacao genotype, subjected to different doses of Mn, Cd, and Mn+Cd in soil, through physiological, biochemical, molecular, and micromorphological and ultrastructural changes. High soil Mn doses favored the maintenance and performance of adequate photosynthetic processes in cacao. However, high doses of Cd and Mn+Cd in soil promoted damage to photosynthesis, alterations in oxidative metabolism, and the uptake, transport, and accumulation of Cd in roots and leaves. In addition, high Cd concentrations in roots and leaf tissues caused irreversible damage to the cell ultrastructure, compromising cell function and leading to programmed cell death. However, there was a mitigation of Cd toxicity when cacao was grown in soils with low Cd doses and in the presence of Mn. Thus, damage to the root and leaf tissues of cacao caused by Cd uptake from contaminated soils can be attenuated or mitigated by the presence of high Mn doses in soil.

Soil flooding and its outcome on cadmium and nutrient uptake affect photosynthetic activity in Inga laurina plants.

Urban areas next to mangroves are subject to progressive heavy metal contamination. Treelets of Inga laurina were collected in this ecosystem and cultivated for 30 days in waterlogged conditions and closed pots (WC) and at field capacity (FC), while exposed to different Cd concentrations (0, 50 and 100 mg·kg-1). Soil water conditions did not affect total Cd in plants, with Cd accumulating in roots and WC inducing less leaf chlorophyll while increasing carotenoids and chlorophyll ratio. Higher net photosynthesis, stomatal conductance, transpiration, and Ci/Ca ratio were observed under the highest Cd concentration and WC, while being conservative in water consumption as shown by the reduction in both water use efficiencies. Nutritional uptake behaved differently for each element, with N, Mg and Ca not being affected by Cd under WC but K increasing with Cd. At FC, plants showed higher values than WC, with the highest Cd concentration at FC showing the highest values overall. Nutrient allocation in organs was affected by WC in N, Mg, K, P and Ca but only P by Cd, with WC reducing nutrients overall and N, Mg and Ca behaving the same in both soil conditions while K was lowered in leaves and increased in roots under WC. P allocation under WC was not hindered by Cd even showing higher values in it than FC in some treatments. Altogether, results indicate that I. laurina can be considered a Cd-tolerant species, especially in WC and it presents a potential to be used as a phytoremediator plant.

Chemical Composition and Biological Activities of Metania and Drulia (Metaniidae) Freshwater Sponges from Amazonia.

Sponges from freshwater environments, unlike marine's, are poorly known producers of natural compounds with medicinal purposes. Amazonian sponges produce massive large specimens and are widely spread, taxonomically diverse and their metabolites could represent a new frontier on unusual natural products to treat diseases such as Alzheimer's and Malaria. Species of Metania and Drulia (Metaniidae) genera are major contributors to the fauna of Amazonian freshwater sponges. Methanolic extracts from several species from these genera had their inhibitory activities evaluated in vitro, for parasite Plasmodium falciparum and acetyl and butyrylcholinesterase enzymes (AChE and BChE). All extracts were able to inhibit AChE, although no activity was observed towards BChE. Drulia uruguayensis extract was the most potent, inhibiting AChE with IC50 =1.04 mg/mL. For antiplasmodial activity, all species showed inhibition to P. falciparum, but Metania reticulata being the most efficient with IC50 =2.7 µg/mL. Mass spectrometry analyses evidenced the presence of fatty acids and sterols in active extracts.

Mechanical stress caused by wind on leaves of Theobroma cacao: Photosynthetic, molecular, antioxidative and ultrastructural responses.

Theobroma cacao is cultivated in the shade, in a so-called 'Cabruca' system, in intercropped with Erithryna or other tree species of economic value, and in full sun as a monoculture in irrigated or chemically-irrigated systems. Since it is a species quite intolerant to wind, it is practically impossible to implant cacao crops under full exposure to the sun, or in areas of frequent winds, without the protection of windbreaks, using arboreal species around the area of culture in the form of box. Wind can cause mechanical stimuli in plants, affecting their growth and development. The objective of this work was to evaluate the photosynthetic changes in mature leaves and the molecular, biochemical and ultrastructural changes in young and mature leaves of the CCN 51 cloned genotype of T. cacao subjected to intermittent (IW) and constant (CW) wind, with velocities of 2.5, 3.5 and 4.5 m s-1, during 3, 6 and 12 h of exposure. It was verified that CW and IW, considering different exposure times, interfered directly in stomatal conductance (gs), transpiration (E) and water use efficiency (WUE), causing a reduction of the photosynthetic rate (A) in mature leaves. In addition, the pulvinus and blade of young and mature leaves, exposed to IW and CW with different exposure times (3 and 12 h), showed marked macroscopic and microscopic mechanical injuries resulting from the constant leaf movement. At both speeds, there was rupture of the cell nuclear membrane in pulvinus and the mesophyll tissues, mainly in the young leaves. On the other hand, in young and mature leaves exposed to CW and IW at different speeds and exposure times, there was lipid peroxidation, increased activity of guaiacol (GPX) and ascorbate (APX) peroxidases in most treatments; and altered expression of transcripts of psba and psbo genes related to the phothosynthetic apparatus and Cu-Zn-sod and per genes related to antioxidative enzymes at the rate of 4.5 m s-1. Younger leaves were more intolerant to mechanical stress caused by the wind, since presented greater macro and microscopic damages and, consequently, greater molecular, biochemical and ultrastructural changes. High wind speeds can seriously compromise the development of young leaves of T. cacao plants and affect their productivity.

Path analysis of phenotypic traits in young cacao plants under drought conditions.

Drought is worldwide considered one of the most limiting factors of Theobroma cacao production, which can be intensified by global climate changes. In this study, we aimed to investigate the phenotypic correlation among morphological characteristics of cacao progenies submitted to irrigation and drought conditions and their partitions into direct and indirect effects. Path analysis with phenotypic plasticity index was used as criteria for estimation of basic and explanatory variables. The experiment was conducted in a greenhouse at the Cacao Research Center (CEPEC), Ilhéus, Bahia, Brazil, in a randomized block 21 x 2 factorial arrangement [21 cacao progenies obtained from complete diallel crosses and two water regimes (control and drought)] and six replications. In general, drought conditions influenced biomass production in most progenies, causing significant reductions in total leaf area, leaf number, leaf biomass, fine-roots length (diameter <1 mm), root volume and root area for considered drought intolerant. All progenies showed alterations in growth due to drought. Phenotypic plasticity was most strongly pronounced in root volume. Stem and root diameters, as well as stem dry biomass were the growth variables with the greatest direct effects on root volume under drought conditions, these characters being indicated in screening of cacao progenies drought tolerant.

Differential accumulation of flavonoids and phytohormones resulting from the canopy/rootstock interaction of citrus plants subjected to dehydration/rehydration.

Water scarcity can elicit drastic changes in plant metabolic and hormonal regulation, which may be of fundamental importance to stress tolerance. The study of plant the metabolic alterations in response to water deficit, especially the effects of the rootstocks level, is important to elucidate the mechanisms associated to drought tolerance. To verify the influence of rootstock and grafting on the tolerance to drought in citrus plants, we analyzed the growth, phytohormone levels and flavonoid profiles in grafted and ungrafted citrus plants subjected to different soil water regimes on plant status (well-watered, moderate drought and severe drought and rehydrated) under field conditions. The experiments were conducted under field conditions in the Brazilian Agricultural Research Corporation (EMBRAPA), Cruz das Almas, BA, Brazil. Water deficit reduced the total leaf area per plant in all canopy/rootstock combinations. Self-grafting reduce root volume, area and length when compared to ungrafted plants. Drought-induced increases in salicylic acid and abscisic acid associated with concomitant reductions in indoleacetic acid were observed in most canopy/rootstock combinations. However, plants with 'Sunki Maravilha' rootstocks exhibited the most pronounced changes in hormonal levels upon drought stress. Associated to these hormonal changes, drought also significantly affected flavonoid content and profile in both leaves and roots of the distinct citrus combinations. Glycosylated (GFs) and polimethoxylated flavonoids were predominantly found in leaves, whereas prenylated coumarins were found in the roots. Leaf levels of GFs (vicenin, F11, rutin and rhoifolin) were particularly modulated by drought in plants with 'Rangpur Santa Cruz' lime rootstock, whereas root levels of prenylated coumarins were most regulated by drought in plants with the 'Sunki Maravilha' root system. Taken together, these data indicate that the impacts of water deficit restriction on growth, hormonal balance and flavonoid profiles significantly varies depending on the canopy/rootstock combinations.

Antimicrobial activity of allylic thiocyanates derived from the Morita-Baylis-Hillman reaction.

Bacterial resistance to commonly used antibiotics has been recognized as a significant global health issue. In this study, we carried out the screening of a family of allylic thiocyanates for their action against a diversity of bacteria and fungi with a view to developing new antimicrobial agents. Allylic thiocyanates bearing halogenated aryl groups, which were readily obtained in two steps from the Morita-Baylis-Hillman adducts, showed moderate-to-high activity against selective pathogens, including a methicillin-resistant S. aureus (MRSA) strain. In particular cases, methyl (Z)-3-(2,4-dichlorophenyl)-2-(thiocyanomethyl)-2-propenoate exhibited antimicrobial activity comparable to the reference antibiotic Imipenem.

Molecular, physiological and biochemical responses of Theobroma cacao L. genotypes to soil water deficit.

Six months-old seminal plants of 36 cacao genotypes grown under greenhouse conditions were subjected to two soil water regimes (control and drought) to assess, the effects of water deficit on growth, chemical composition and oxidative stress. In the control, soil moisture was maintained near field capacity with leaf water potentials (ΨWL) ranging from -0.1 to -0.5 MPa. In the drought treatment, the soil moisture was reduced gradually by withholding additional water until ΨWL reached values of between -2.0 to -2.5 MPa. The tolerant genotypes PS-1319, MO-20 and MA-15 recorded significant increases in guaiacol peroxidase activity reflecting a more efficient antioxidant metabolism. In relation to drought tolerance, the most important variables in the distinguishing contrasting groups were: total leaf area per plant; leaf, stem and total dry biomass; relative growth rate; plant shoot biomass and leaf content of N, Ca, and Mg. From the results of these analyses, six genotypes were selected with contrasting characteristics for tolerance to soil water deficit [CC-40, C. SUL-4 and SIC-2 (non-tolerant) and MA-15, MO-20, and PA-13 (tolerant)] for further assessment of the expression of genes NCED5, PP2C, psbA and psbO to water deficit. Increased expression of NCED5, PP2C, psbA and psbO genes were found for non-tolerant genotypes, while in the majority of tolerant genotypes there was repression of these genes, with the exception of PA-13 that showed an increased expression of psbA. Mutivariate analysis showed that growth variables, leaf and total dry biomass, relative growth rate as well as Mg content of the leaves were the most important factor in the classification of the genotypes as tolerant, moderately tolerant and sensitive to water deficit. Therefore these variables are reliable plant traits in the selection of plants tolerant to drought.

Antimicrobial activity of allylic thiocyanates derived from the Morita-Baylis-Hillman reaction

Bacterial resistance to commonly used antibiotics has been recognized as a significant global health issue. In this study, we carried out the screening of a family of allylic thiocyanates for their action against a diversity of bacteria and fungi with a view to developing new antimicrobial agents. Allylic thiocyanates bearing halogenated aryl groups, which were readily obtained in two steps from the Morita-Baylis-Hillman adducts, showed moderate-to-high activity against selective pathogens, including a methicillin-resistant S. aureus (MRSA) strain. In particular cases, methyl (Z)-3-(2,4-dichlorophenyl)-2-(thiocyanomethyl)-2-propenoate exhibited antimicrobial activity comparable to the reference antibiotic Imipenem.

Virulence factors associated with pediatric shigellosis in Brazilian Amazon.

Shigellosis is a global human health problem and the incidence is highest among children. In the present work, main Shigella virulence genes was examined by PCR and compared to symptoms of pediatric shigellosis. Thirty Shigella isolates were identified from an etiologic study at which 1,339 children ranging 0-10 years old were enrolled. S. flexneri was the most frequent species reaching 60.0% of isolates, 22.2% were S. sonnei, and 6.6% were both S. dysenteriae and S. boydii. All Shigella infected children had diarrhea, but not all were accompanied by others symptoms of bacillary dysentery. Among major virulence genes, the PCR typing revealed ipaBCD was present in all isolates, followed by IpaH7.8, set-1A, set-1B, sen/ospD3, virF, and invE. The pathogenic potential of the ShET-1B subunit was observed in relation to dehydration (P < 0.001) and ShET-2 related to the intestinal injury (P = 0.033) evidenced by the presence of bloody diarrhea. Our results show associations among symptoms of shigellosis and virulence genes of clinical isolates of Shigella spp.

Comparison of the performance of two spontaneous sedimentation techniques for the diagnosis of human intestinal parasites in the absence of a gold standard.

Performance evaluation of diagnostic tests is critical in the search for accurate diagnoses. A gold standard test is usually absent in parasitology, thus rendering satisfactory assessment of diagnostic accuracy difficult. Moreover, reliability (assessed by the study of repeatability) is a rarely studied characteristic of diagnostic tests. This study compared and evaluated the performance (repeatability, concordance and accuracy) of the spontaneous sedimentation technique (SST) and the Paratest for the diagnosis of Giardia lamblia, Entamoeba histolytica complex, Blastocystis spp., Ascaris lumbricoides, hookworm, Trichuris trichiura and Calodium hepaticum. Fecal samples of 143 individuals were separated into three replicates for each test. Concordance and homogeneity of the results between replicates of each test and between tests were evaluated. Proportions of positives, sensitivity and specificity were estimated using a Bayesian Latent Class Model. High repeatability of both tests was found for the detection of intestinal parasites, except for Blastocystis spp. and hookworm. Concordance between tests was generally high (concordance correlation coefficient, 0.72-0.88), except for Blastocystis spp., hookworm and T. trichiura. The Paratest detected more cases of Blastocystis spp. and fewer of hookworm than the SST. The tests were quite discordant in the detection of T. trichiura. A low sensitivity (39.4-49.2% for SST, 35.8-53.8% for Paratest) and a high specificity (93.2-97.2%) were found for both tests. The Paratest presented a slightly higher sensitivity for the diagnosis of Blastocystis spp. (53.8%), and SST did so for hookworm (49.2%). This is the first study on repeatability and accuracy (using a Bayesian approach) of two spontaneous sedimentation techniques. These results suggest underdiagnosis of little dense parasitic forms due to technical limitations in both tests. We conclude that the combined study of repeatability, concordance and accuracy is a key strategy for better evaluation of the performance of tests and is also useful for the identification of technical limitations.

Calodium hepaticum: household clustering transmission and the finding of a source of human spurious infection in a community of the Amazon region.

BACKGROUND: Calodium hepaticum (syn. Capillaria hepatica) is a worldwide helminth parasite of which several aspects of transmission still remain unclear. In the Amazon region, the mechanism of transmission based on the ingestion of eggs present in the liver of wild mammals has been suggested as the cause of the spurious infections described. We performed an epidemiological investigation to determine the incidence, risk of spurious infection and the dynamics of transmission of C. hepaticum in a community of the Brazilian Amazon. METHODOLOGY/PRINCIPAL FINDINGS: Stool samples of 135 individuals, two dog feces and liver tissue from a peccary (captured and eaten by the residents) were analyzed by conventional microscopy. Dog feces were collected from the gardens of households presenting human cases of spurious C. hepaticum infections. Community practices and feeding habits related to the transmission of the parasite were investigated. The individual incidence of spurious infection was 6.7% (95% CI: 2.08-11.24). Cases of spurious infection were observed in 7.5% of the families and the household incidence was from 50% to 83.3%. The risk of spurious infection was 10-fold greater in persons consuming the liver of wild mammals (p = 0.02). The liver tissue of a peccary and one feces sample of a dog presented eggs of C. hepaticum. The consumption of the infected liver was the cause of the spurious infections reported in one household. CONCLUSIONS/SIGNIFICANCE: This is the first identification of a source of spurious infection by C. hepaticum in humans and we describe a high rate of incidence in household clusters related to game liver alimentary habits. The finding of a dog feces contaminating peridomiciliary ground suggests the risk of new infections. We conclude that the mechanism of transmission based on the ingestion of liver is important for the dynamics of transmission of C. hepaticum in the studied area.