Active and exo-site inhibition of human factor Xa: Structure of des-Gla factor Xa inhibited by NAP5, a potent nematode anticoagulant protein from Ancylostoma caninum

Hookworms are hematophagous nematodes capable of growth, development and subsistence in living host systems such as humans and other mammals. Approximately one billion, or one in six, people worldwide are infected by hookworms causing gastrointestinal blood loss and iron deficiency anemia. The hematophagous hookworm Ancylostoma caninum produces a family of small, disulfide-linked protein anticoagulants (75-84 amino acid residues). One of these nematode anticoagulant proteins, NAP5, inhibits the amidolytic activity of factor Xa (fXa) with K-i = 43 pM, and is the most potent natural fXa inhibitor identified thus far. The crystal structure of NAP5 bound at the active site of gamma-carboxyglutamic acid domainless factor Xa (des-fXa) has been determined at 3.1 angstrom resolution, which indicates that Asp189 (fXa, S1 subsite) binds to Arg40 (NAP5, P1 site) in a mode similar to that of the BPTI/trypsin interaction. However, the hydroxyl group of Ser39 of NAP5 additionally forms a hydrogen bond (2.5 angstrom) with His57 NE2 of the catalytic triad, replacing the hydrogen bond of Ser195 OG to the latter in the native structure, resulting in an interaction that has not been observed before. Furthermore, the C-terminal extension of NAP5 surprisingly interacts with the fXa exosite of a symmetry-equivalent molecule forming a short intermolecular beta-strand as observed in the structure of the NAPc2/fXa complex. This indicates that NAP5 can bind to fXa at the active site, or the exosite, and to fX at the exosite. However, unlike NAPc2, NAP5 does not inhibit fVIIa of the fVIIa/TF complex. (c) 2007 Elsevier Ltd. All rights reserved.

Medicinal plants' hosting ability for nematode meloidogyne javanica and m. incognita

Baida, F.C., D.C. Santiago, L. H. I. Vidal, L. C. Baida, C. T. Stroze. 2011. Medicinal Plants' Hosting Ability for Nematode Suitability Meloidogyne incognita and M. javanica. Nematropica 41: 150-153.Medicinal plants can be attacked by pests, diseases and nematodes, which can compromise the quality and quantity of their healing properties and their yield. The aim of this study was to analyse the reaction of 15 medicinal plant species to the nematode Meloidogyne spp. The seedlings were produced by seed germination or cuttings under greenhouse conditions. A completely random experimental pattern of 15 treatments and 10 replications was chosen for the study. The seedlings were inoculated with approximately 5000 eggs + J(2)/plant 20 days after planting. Plant height and fresh and dry leaf weight, were measured 60 days after planting. The roots were collected, thoroughly washed and stained with Philoxine B and then processed to extract the eggs to determine the reproduction factor. For M. incognita the results showed that Chamomile was susceptible with RF = 1,64 making it a good host, and the other plants were resistant (RF < 1), and for M. javanica that all the plants showed resistance (RF < 1), Myrrh, Rue and Balsam demonstrating immunity (RF = 0).

Predation potential and biology of Protogamasellopsis posnaniensis Wisniewski & Hirschmann (Acari: Rhodacaridae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Helminthological records of six-banded Armadillos Euphractus sexcinctus (Linnaeus, 1758) from the Brazilian semi-arid region, Patos county, Paraiba state, including new morphological data on Trichohelix tuberculata (Parona and Stossich, 1901) Ortlepp, 1922 and proposal of Hadrostrongylus ransomi nov comb.

This work aimed to evaluate the gastrointestinal helminthfauna composition of six-banded armadillos from the Brazilian semi-arid region. Gastrointestinal contents of six road-killed adult animals from Patos County, Paraiba State, were analyzed. Six species of nematodes, comprising five genera and four families, were recovered from the analyzed animals. New morphological data on Trichohelix tuberculata is given, along with a new taxonomical proposal for Hadrostrongylus ransomi (Travassos, 1935) n. comb. This is the first record for parasitic helminths in this host from the Brazilian semi-arid.

Helminth parasites of Leptodactylus podicipinus (Anura: Leptodactylidae) from south-eastern Pantanal, State of Mato Grosso do Sul, Brazil

Forty-three specimens of Leptodactylus podicipinus (Anura: Leptodactylidae) were collected in the south-eastern Pantanal, municipality of Corumba, State of Mato Grosso do Sul, Brazil in February and July 2007, and examined for endoparasites. Forty (93%) specimens were infected with at least one helminth species. The predominant parasites were nematodes (Aplectana sp., Cosmocerca podicipinus, Oswaldocruzia lopesi, Physalopteroides venancioi, Rhabdias sp.), but the trematode Catadiscus propinquus also showed high prevalence. The trematodes Infidum infidum and Travtrema stenocotyle were also found, but in only one specimen. Adult frogs showed higher parasite diversity than subadults. Leptodactylus podicipinus was preferentially infected by direct life-cycle parasites and was reported as a new host record for seven helminth species.

Efeito alelopático de extrato aquoso de folhas de Azadirachta indica A. Juss. em alface, soja, milho, feijão e picão-preto

The family Meliaceae has been identified as one of the most promising for biological control, as most species have biologically active compounds. Among the species, the Neem (Azadirachta indica A. Juss) is that stands out most for its efficiency and low toxicity, and it is utilized to control insects, fungi and nematodes. The aim of this work was to verify the allelopathic potential of aqueous extract of fresh leaves of Neem on germination of lettuce, soybeans, maize, beans and Bidens pilosa, as a way to control this weed. The extract was obtained by grinding 200g/L of fresh leaves of the Neem (considered extract 100%) and diluting in distilled water at concentrations of 80, 60, 40, and 20%, and a control with only water. The seeds were kept in the germination chamber at a temperature of 25 degrees C, with photoperiod of 12 hours of light. The aqueous extract affected the germination percentage and average length root of lettuce, with values of 1% and 0.1, cm respectively. For the soybeans, the extract influenced negatively the average time of germination and average speed of germination. There was a drastic reduction of the average root length at higher concentration for soybean, corn and beans. For Bidens pilosa all the parameters analyzed were negatively affected, which means sensitivity to the extract. The bioassay performed in the laboratory showed that the aqueous extract of fresh leaves of Azadirachta indica has allelopathic effect on all species.

Fatores de riscos associados à ocorrência de helmintos gastrintestinais em cabras leiteiras mantidas em sistema de pastejo

Infection due to the gastrointestinal nematodes can negatively interact with the productive performance of dairy goats, and in some cases can induce mortality in the flocks. The objective was to study the influence of season, parturition order, postpartum, lactation and breed standard on the population of helminths in dairy goats. 31 goats were used, with 15 Saanen and 16 F1 (1/2 Boer + 1/2 Saanen). of these, 13 were from the 1st lactation and 18 from the 2nd or more lactations. The highest fecal egg counts occurred during the summer and spring, with FEC in Saanen higher (p < 0.01) than in F1 animals (1/2 Boer + 1/2 Saanen). The animals with 1st lactation showed higher FEC (p < 0.004) than the animals in 2nd or more lactation. During the peripartum the largest FEC for both breeds standards were in the week of the birth (p < 0.05), followed by post-partum and pre-partum. In peripartum the Saanen goats showed higher FEC (p < 0.02) than the F1 and primiparous (p < 0.008) in relation to pluriparous. F1 animals are resistant to helminthes when compared to Saanen animals, demonstrating that crossing of resistant breeds with high production breeds is a viable alternative to increase productivity without compromising the parasitary stability of the herd. The peripartum period is a risk factor associated to the occurrence of gastrointestinal helminths, especially in Saanen goats from first lactation.

Avaliação Comparativa da Agressividade de Meloidogyne javanica e M. incognita à Variedade SP 911049 de Cana-de-açúcar

Nematodes cause extensive losses to sugarcane in Brazil and also in other producing regions. Meloidogyne incognita, M. javanica and Pratylenchus zeae are the key species for this culture worldwide. In the present study, the aggressiveness of M. javanica and M. incognita to sugarcane variety SP 911049 was evaluated comparatively,. The following parameters were compared: reproduction factor (RF) of these nematodes, effect of nematodes in the natural incidence of pests, and the influence on the development and technological characteristics of sugarcane. Considering the data of RF, biometrics, natural infestation of pests, mortality of plants, and technological variables, it was concluded that M. javanica was more aggressive to sugarcane, although its rate of multiplication was much smaller than the one of M. incognita.

Reprodução de Meloidogyne incognita, M. javanica e Pratylenchus coffeae em Diferentes Cultivares de Bananeira

Banana (Musa spp.) is one of the most consumed fruits in the world, and it is cultivated in many tropical countries. In Brazil, the productivity is low due to several factors such as the incidence of pests and diseases. The nematodes are among the most important problems found in the production of bananas. This research studied the reproduction of Meloidogyne incognita race 2, M. javanica and Pratylenchus coffeae in different cultivars of banana. The evaluation of the experiments with Meloidogyne spp. was carried out 120 days after the inoculation. The analyzed parameters were: number of gall, number of the external masses of eggs, number of eggs per gram of root, reproduction factor and number of juveniles in the soil. The evaluation of the experiment with P. coffeae was carried out 90 days after the inoculation; the final population was evaluated. The cultivars PV-0344, Maca, Grande Naine, FHIA 01, Thap Maeo and Prata Ana allowed greater multiplication of P. coffeae than Caipira, SH-3640 and FHIA-18. For M. javanica, the cultivars Calypso, Bucanneer, Grande Naine, PV-0344, Nanicao Magario, FHIA-02, SH-3640, Pacovan, and Prata Ana exhibited larger populations, decreasing in Maca. All cultivars allowed high multiplication rates of M. incognita.

Aspects of IFP for citrus production in Brazil

Brazilian citriculture represents about 25% of the total world citrus production with an area of 851,518 ha and a total production of more than 17 million tons in 1996. Besides its importance to the brazilian economy, represented by more than 1 billion US $ by year from FCOJ exportation, the citriculture has problems related to low productivity, due to several cultural practices and management. The productivity would be improved by an IFP system. The main problems are related to soil, as poor conservation, use of poor drained soils and bad preparation for planting; diseases-canker, CVC, leprosis, Phytophtora gummosis and other fungus diseases; pests - mites, scales, nematodes and others; the use of disease free and improved scion and rootstocks propagative material is usual; cultural practices - as nutrition, irrigation, wind breaks, weed control, pruning, replant and others, as density planting could be improved. Some possible solutions will be discussed for improving the brazilian citrus productivity and quality by IFP, based on research made.

Metabolismo oxidativo de neutrófilos em ovelhas naturalmente infectadas por nematódeos gastrintestinais e correlação entre nível sérico de cortisol e carga parasitária

The relationships between neutrophils oxidative metabolism, cortisol serum levels and worm burden, estimated by fecal egg count (EPG), were studied in sheep naturally infected by gastrointestinal parasites at the end of pregnancy, during lactation, and after weaning. Twenty-two Suffolk sheep three to four year old, of same parity and season of parturition were used. Serum cortisol was determined by radioimmunoassay and the neutrophils oxidative metabolism by the nitroblue tetrazolium (NBT) reduction test. The highest EPG values were observed on the fifth week of lactation while the highest rates of cortisol and reduction of NBT occurred on the fourth week after weaning. A positive correlation (r = 0.52; P<0.01) was observed between the neutrophils capacity to reduce the NBT and the serum cortisol concentration in the pre-parturition period. Otherwise, the neutrophils oxidative metabolism decreased near to the parturition. A negative correlation (r = -0.39; P<0.01) between EPG and NBT reduction test was observed after weaning, which was coincident with the increase in the neutrophils capacity to reduce NBT, indicating that animals showing better immune response after weaning have neutrophils with higher oxidative metabolism and lower worm burden.

Caracterização das fases de desenvolvimento de células gigantes induzidas por Meloidogyne exigua em raiz de seringueira (Hevea brasiliensis Muell. Arg.)

The developmental phases of giant cells induced by root-knot nematodes (Meloidogyne exigua) in rubber plant (Hevea brasiliensis) root were studied in relation to its number and size evaluated in eight sample dates. The results were subject to cluster analysis and principal component analysis. Sample dates were clearly distinct regarding giant cell development. As a result, the nematode infestation cycle was characterized by the following sequential phases: initial, equilibrium, choice and final.

Resistance of Santa Inês and Ile de France suckling lambs to gastrointestinal nematode infections.

A trial was carried out to determine the resistance to natural infection by gastrointestinal nematodes in 12 Santa Inês and nine Ile de France lambs before weaning. Faecal samples were obtained for faecal nematode egg counts (FEC). Blood samples were collected to determine packed cell volume (PCV), total plasma protein levels and peripheral eosinophil counts. Most Ile de France lambs (77.8%) were treated with an anthelmintic at 43 days of age, while 50% off Santa Inês lambs were treated at weaning, 57 days of age. The mean PCV values were normal in Santa Inês lambs, while in Ile de France lambs showed lower values reaching 22.3% at 43 days of age. The lowest mean plasma protein values were observed in Ile de France lambs (4.13 g/dl) at 43 days of age and in Santa Inês lambs (5.0 g/dl) at 57 days of age. Before weaning, Santa Inês lambs were susceptible to natural infections by gastrointestinal nematodes but with a greater capacity to stand the adverse effects of parasitism compared to Ile de France lambs.

Avaliação do kit TF-Test para o diagnóstico das infecções por parasitas gastrintestinais em ovinos

This study was performed to standardize parasite egg counting in feces of sheep by TF-Test, in addition to compare this test to the Gordon & Whitlock technique (G&W). Twenty-four lambs were artificially infected with Haemonchus contortus throughout 12 weeks. At the end of this time, faecal samples were taken and animals were slaughtered for worm identification and counting. G&W and TF-Test methods were carried out on each fecal sample. Both tests showed Haemonchus eggs in 95.8% of the samples (P>0.05). The correlation coefficients (r) between fecal egg counts (FEC) using G&W × Total Worm Count (TWC) were r=0.52 (not transformed data) and r=0.85 (transformed data); between FEC by TF-Test × TWC were r=0.51 (not transformed data) and r=0.87 (transformed data). Other 100 fecal samples were taken from naturally infected sheep. In these animals, the G&W and TF-Test methods showed 85% and 86% of fecal samples positive for Strongylidea eggs, respectively (P>0.05). Also in those animals, Eimeria oocysts were found in 33% of fecal samples by TF-Test, whereas in the G&W only 12% were positive (P<0.001). For Strongyloides spp., TF-Test showed 15% of positive fecal samples, whereas G&W showed 5% (P<0.05). In conclusion, both methods were efficient to diagnose gastrointestinal nematodes and TF-Test was superior to diagnose oocysts of Eimeria spp. and eggs of Strongyloides spp; conversely, Strongylidea eggs counting using TF-Test was underestimated.