Polyphenol oxidase from yacon roots (Smallanthus sonchifolius)

Polyphenol oxidase (E.C. 1.14.18.1) (PPO) extracted from yacon roots (Smallanthus sonchifolius) was partially purified by ammonium sulfate fractionation and separation on Sephadex G-100. The enzyme had a molecular weight of 45 490+/-3500 Da and Km values of 0.23, 1.14, 1.34, and 5.0 mM for the substrates caffeic acid, chlorogenic acid, 4-methylcatechol, and catechol, respectively. When assayed with resorcinol, DL-DOPA, pyrogallol, protocatechuic, p-coumaric, ferulic, and cinnamic acids, catechin, and quercetin, the PPO showed no activity. The optimum pH varied from 5.0 to 6.6, depending on substrate. PPO activity was inhibited by various phenolic and nonphenolic compounds. p-Coumaric and cinnamic acids showed competitive inhibition, with Ki values of 0.017 and 0.011 mM, respectively, using chlorogenic acid as substrate. Heat inactivation from 60 to 90 degrees C showed the enzyme to be relatively stable at 60-70 degrees C, with progressive inactivation when incubated at 80 and 90 degrees C. The Ea (apparent activation energy) for inactivation was 93.69 kJ mol-1. Sucrose, maltose, glucose, fructose, and trehalose at high concentrations appeared to protect yacon PPO against thermal inactivation at 75 and 80 degrees C.     

Occurrence of nematodes and anthelmintic management of ostrich farms from different Brazilian states: Libyostrongylus douglassii dominates mixed infections

Ostriches were imported to Brazil in the early 1990s without appropriate parasite control leading to a co-importation of parasites. Libyostrongylus douglassii, Libyostrongylus dentatus and Codiostomum struthionis are nematodes of the ostriches that have been reported on some Brazilian farms but no national survey has been performed. Moreover, little is known about anthelmintic usage to control these parasites in Brazil. Therefore, the occurrence of L. douglassii, L. dentatus and C. struthionis and the anthelmintic management practices in Brazilian ostrich farms were studied. A questionnaire about management practices and information on how to collect, store and ship feces to the laboratory were sent to 17 farms located in 9 Brazilian states. Received fecal samples were submitted to fecal cultivation and larval harvested. Infective larvae were morphologically analyzed and quantified. The majority of the farms (88%) used an anthelmintic at least once a year. Ivermectin was the principal anthelmintic. Libyostrongylus sp. and C. struthionis were found in samples from 76 and 18% of the examined farms, respectively. The presence of L. douglassii (61-97%) was higher than L. dentatus (3-57%) and C. struthionis (2-30%). At the farm level, both species of Libyostrongylus were always found together. Sheath tail measurements of their infective larvae were crucial for morphological diagnosis. L. dentatus and L. douglassii are well adapted to Brazil and are widely distributed in the Brazilian territory, proving that this mixed infection in Brazilian ostrich's farms is common.     

Draft genome sequence of the sexually transmitted pathogen Trichomonas vaginalis

We describe the genome sequence of the protist Trichomonas vaginalis, a sexually transmitted human pathogen. Repeats and transposable elements comprise about two-thirds of the approximately 160-megabase genome, reflecting a recent massive expansion of genetic material. This expansion, in conjunction with the shaping of metabolic pathways that likely transpired through lateral gene transfer from bacteria, and amplification of specific gene families implicated in pathogenesis and phagocytosis of host proteins may exemplify adaptations of the parasite during its transition to a urogenital environment. The genome sequence predicts previously unknown functions for the hydrogenosome, which support a common evolutionary origin of this unusual organelle with mitochondria.     

Oxidative response of neutrophils to platelet-activating factor is altered during acute ruminal acidosis induced by oligofructose in heifers

Reactive oxygen species (ROS) production is one of the main mechanisms used to kill microbes during innate immune response. D-lactic acid, which is augmented during acute ruminal acidosis, reduces platelet activating factor (PAF)-induced ROS production and L-selectin shedding in bovine neutrophils in vitro. This study was conducted to investigate whether acute ruminal acidosis induced by acute oligofructose overload in heifers interferes with ROS production and L-selectin shedding in blood neutrophils. Blood neutrophils and plasma were obtained by jugular venipuncture, while ruminal samples were collected using rumenocentesis. Lactic acid from plasma and ruminal samples was measured by HPLC. PAF-induced ROS production and L-selectin shedding were measured in vitro in bovine neutrophils by a luminol chemiluminescence assay and flow cytometry, respectively. A significant increase in ruminal and plasma lactic acid was recorded in these animals. Specifically, a decrease in PAF-induced ROS production was observed 8 h after oligofructose overload, and this was sustained until 48 h post oligofructose overload. A reduction in PAF-induced L-selectin shedding was observed at 16 h and 32 h post oligofructose overload. Overall, the results indicated that neutrophil PAF responses were altered in heifers with ruminal acidosis, suggesting a potential dysfunction of the innate immune response.     

A mathematical approach to simulate spatio-temporal patterns of an insect-pest,the corn rootworm Diabrotica speciosa (Coleoptera: Chrysomelidae) in intercropping systems

We report on the use of a spatially explicit model and clustering analysis in order to investigate habitat manipulation as a strategy to regulate natural population densities of the insect-pest Diabrotica speciosa. Habitat manipulation involved four major agricultural plants used as hosts by this herbivore to compose intercropping landscapes. Available biological parameters for D. speciosa on bean, soybean, potato and corn obtained under laboratory conditions were used to group the homogeneous landscapes, composed by each host plant, by a similarity measure of host suitability either for larval survival and development, and adult survival and fecundity. The results pointed corn as the most dissimilar culture. Therefore, intercropping corn with any other crop system tested could reduce insect spread through landscape. This was proved using a cellular automata model which simulate the physiological and behavioural traits of this insect, and also different spatial configurations of the intercropping. Spatio-temporal patterns obtained by simulations demonstrated that the availability of corn bordering the field edge, which are areas more likely to invasion, is key for insect population control.     

Differentiation in light energy dissipation between hemiepiphytic and non-hemiepiphytic Ficus species with contrasting xylem hydraulic conductivity

Hemiepiphytic Ficus species (Hs) possess traits of more conservative water use compared with non-hemiepiphytic Ficus species (NHs) even during their terrestrial growth phase, which may result in significant differences in photosynthetic light use between these two growth forms. Stem hydraulic conductivity, leaf gas exchange and chlorophyll fluorescence were compared in adult trees of five Hs and five NHs grown in a common garden. Hs had significantly lower stem hydraulic conductivity, lower stomatal conductance and higher water use efficiency than NHs. Photorespiration played an important role in avoiding photoinhibition at high irradiance in both Hs and NHs. Under saturating irradiance levels, Hs tended to dissipate a higher proportion of excessive light energy through thermal processes than NHs, while NHs dissipated a larger proportion of electron flow than Hs through the alternative electron sinks. No significant difference in maximum net CO2 assimilation rate was found between Hs and NHs. Stem xylem hydraulic conductivity was positively correlated with maximum electron transport rate and negatively correlated with the quantum yield of non-photochemical quenching across the 10 studied Ficus species. These findings indicate that a canopy growth habit during early life stages in Hs of Ficus resulted in substantial adaptive differences from congeneric NHs not only in water relations but also in photosynthetic light use and carbon economy. The evolution of epiphytic growth habit, even for only part of their life cycle, involved profound changes in a suite of inter-correlated ecophysiological traits that persist to a large extent even during the later terrestrial growth phase.     

Nitrogen oxides emission from soils bearing a potato crop as influenced by fertilization with treated pig slurries and composts

Nitrous oxide, nitric oxide and denitrification losses from an irrigated soil amended with organic fertilizers with different soluble organic carbon fractions and ammonium contents were studied in a field study covering the growing season of potato (Solanum tuberosum). Untreated pig slurry (IPS) with and without the nitrification inhibitor dicyandiamide (DCD), digested thin fraction of pig slurry (DTP), composted solid fraction of pig slurry (CP) and composted municipal solid waste (MSW) mixed with urea were applied at a rate of 175 kg available N ha⁻¹, and emissions were compared with those from urea (U) and a control treatment without any added N fertilizer (Control). The cumulative denitrification losses correlated significantly with the soluble carbohydrates, dissolved N and total C added. Added dissolved organic C (DOC) and dissolved N affected the N₂O/N₂ ratio, and a lower ratio was observed for organic fertilizers than from urea or unfertilized controls. The proportion of N₂O produced from nitrification was higher from urea than from organic fertilizers. Accumulated N₂O losses during the crop season ranged from 3.69 to 7.31 kg N₂O-N ha⁻¹ for control and urea, respectively, whereas NO losses ranged from 0.005 to 0.24 kg NO-N ha⁻¹, respectively. Digested thin fraction of pig slurry compared to IPS mitigated the total N₂O emission by 48% and the denitrification rate by 33%, but did not influence NO emissions. Composted pig slurry compared to untreated pig slurry increased the N₂O emission by 40% and NO emission by 55%, but reduced the denitrification losses (34%). DCD partially inhibited nitrification rates and reduced N₂O and NO emissions from pig slurry by at least 83% and 77%, respectively. MSW+U, with a C:N ratio higher than that of the composted pig slurry, produced the largest denitrification losses (33.3 kg N ha⁻¹), although N₂O and NO emissions were lower than for the U and CP treatments.This work has shown that for an irrigated clay loam soil additions of treated organic fertilizers can mitigate the emissions of the atmospheric pollutants NO and N₂O in comparison with urea.     

Resistance to <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> race 1 in the lettuce cultivars Grand Rapids and Salinas-88

The objective of this work was to check the possible allelism between two sources of resistance to the root-knot nematode Meloidogyne incognita race 1 in lettuce (‘Grand Rapids’ and ‘Salinas-88’). The experiments were carried out in greenhouses, in expanded 128-cell polystyrene trays filled with commercial substrate. Lettuce cultivars ‘Salinas 88’ and ‘Grand Rapids’ were tested along with the populations F₁ (‘Grand Rapids’ × ‘Salinas-88’), F₂ (‘Grand Rapids’ × ‘Salinas-88’), F₃ (‘Grand Rapids’ × ‘Salinas-88’), and with F₄ families derived from the latter population. Seedlings were inoculated 15 days after sowing with a nematode egg suspension equivalent to 30 eggs ml⁻¹ of substrate. Plants were evaluated for apparent gall incidence, gall scores, egg mass scores and extracted egg numbers 45 days after the inoculation date. There was evidence that two different genes are involved in control of resistance to M. incognita race 1 in lettuce cultivars Grand Rapids and Salinas-88. Lines with higher levels of nematode resistance than either Grand Rapids or Salinas-88 could be selected in the F4 generation of the cross between these resistant parental lines.