Hepatic photosensitization in buffaloes intoxicated by Brachiaria decumbens in Minas Gerais state, Brazil.

The aim of this paper is to report the study of hepatogenous photosensitization in buffaloes during two outbreaks provoked by ingestion of Brachiaria decumbens in Minas Gerais state, Brazil. Ten young buffaloes in outbreak 1 and seven buffaloes in outbreak 2 were intoxicated by B. decumbens. Nine clinically healthy buffaloes raised under the same conditions as the sick animals served as the control group. All animals were subjected to clinical examination, and serum was collected to measure gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST), direct bilirubin (DB), indirect bilirubin (IB) and total bilirubin (TB) as indicators of liver function and urea and creatinine as indicators of renal function. Histopathology of liver fragments from five different animals was carried out. During the outbreaks and every two months for one year, samples of grass from paddocks where the animals got sick were collected for quantitative evaluation of the saponin protodioscin, combined with observations of pasture characteristics and daily rainfall. Clinical signs included apathy, weight loss, restlessness, scar retraction of the ears and intense itching at the skin lesions, mainly on the rump, the tail head, neck and hindlimbs, similar to the signs observed in other ruminants. Only the GGT enzyme presented significantly different (P < 0.01) serum levels between intoxicated animals (n = 17) and healthy animals (n = 9), indicating liver damage in buffaloes bred in B. decumbens pastures. Microscopy of the liver showed foamy macrophages and lesions of liver disease associated with the presence of crystals in the bile ducts, which have also been found in sheep and cattle poisoned by grasses of the genus Brachiaria. During the outbreaks, protodioscin levels were higher than 3%, and shortly after, these levels were reduced to less than 0.80%, suggesting a hepatic injury etiology. The outbreaks took place at the beginning of the rainy season, and there was a positive correlation between saponin and the amount of rainfall, as well as between saponin and the amount of green leaves in the pasture. These findings indicate that the grass was more toxic in this period. This is the first report of photosensitization by B. decumbens in buffalo.

Antagonistic activity of the fungus Pochonia chlamydosporia on mature and immature Toxocara canis eggs.

In vitro tests were performed to evaluate the ability of 6 isolates of the nematophagous fungus Pochonia chlamydosporia to infect immature and mature Toxocara canis eggs on cellulose dialysis membrane. There was a direct relationship between the number of eggs colonized and the increase in the days of interaction, as well as between the number of eggs colonized and the increase in the concentration of chlamydospores (P<0.05). Immature eggs were more susceptible to infection than mature eggs. The isolate Pc-04 was the most efficient egg parasite until the 7th day, and showed no difference in capacity to infect mature and immature eggs in comparison to Pc-07 at 14 and 21 days of interaction, respectively. Isolate Pc-04 was the most infective on the two evolutionary phases of the eggs at most concentrations, but its ability to infect immature eggs did not differ from that presented by the isolates Pc-07 and Pc-10 at the inoculum level of 5000 chlamydospores. Colonization of infective larvae inside or outside the egg was observed in treatments with the isolates Pc-03, Pc-04, Pc-07 and Pc-10. The isolate Pc-04 of P. chlamydosporia has great biological capacity to destroy immature and mature T. canis eggs in laboratory conditions.

The biological control of Ancylostoma spp. dog infective larvae by Duddingtonia flagrans in a soil microcosm.

Experiments to evaluate the potential ability of the nematode-trapping fungus Duddingtonia flagrans (Isolate CG768) to prey on the Ancylostoma spp. dog infective larvae (L(3)) in pasteurized soil were performed through several laboratory assays. A microcosm approach was used with increasing fungal concentrations in an inoculum of a chlamydospore water suspension. The highest fungal concentrations provide a more consistent larval reduction than the lowest concentrations, but no difference was observed from 10,000 to 25,000 chlamydospores per grain of soil. When using D. flagrans in a water suspension, in white rice and in milled maize, there were reductions in the larval population of 72.0%, 78.4% and 79.4%, respectively, but there was no difference between white rice and milled maize (p<0.05). To evaluate the nematode control by D. flagrans inoculated in milled maize at 10,000 chlamydospores per grain of soil under greenhouse conditions, observations were performed at 10, 15, 20, 25 and 30 days after inoculation and the percent reduction in the larval population was 61.4%, 73.2%, 70.8%, 64.5% and 57%, respectively (p<0.05). There was an inverse relationship between the number of L(3) recovered from the soil and the total days of exposure to the fungus (p<0.05). These results showed that D. flagrans could present some potential to be used as a non-chemotherapeutic alternative for regulation of Ancylostoma spp. populations in the environment.

Predation of Ancylostoma spp. dog infective larvae by nematophagous fungi in different conidial concentrations.

In the present work, it was evaluated the in vitro effect of 12 isolates from the fungal species Arthrobotrys, Duddingtonia, Nematoctonus and Monacrosporium genera in different conidial concentrations on the capture of Ancylostoma spp. dog infective larvae (L(3)), on 2% water-agar medium at 25 degrees C, at the end of a period of 7 days. The concentrations used for each nematophagous fungus were 1000, 5000, 10,000, 15,000 and 20,000conidia/Petri dish plated with 1000 Ancylostoma spp. L(3). All nematode-trapping fungi isolates tested reduced the averages of the uncaptured Ancylostoma spp. L(3) recovered, with the increase of the fungal inoculum concentration, in comparison to the fungus-free control (p<0.05). The adhesive network producing species were better predators than the constricting ring or adhesive knob producing species. Duddingtonia flagrans (Isolate CG768) was the most effective, reducing the averages of the uncaptured Ancylostoma spp. L(3) recovered in 92.8%, 96.3%, 97.5%, 98.3% and 98.9%, respectively in five fungal inoculum concentrations established. Other effective nematophagous fungi were Arthrobotrys robusta (Isolate I31), which reduced the averages of the uncaptured Ancylostoma spp. L(3) recovered in 85.4%, 88.3%, 90.7%, 92.5% and 95.2%, and Arthrobotrys oligospora (Isolate A183), with reductions of 66.6%, 79.8%, 86.8%, 89.5% and 90.8%, respectively for both, in the five fungal inoculum concentrations established. No difference was found between Isolates A183 and I31 in the conidial concentrations of 15,000/Petri dish. Nematoctonus robustus (Isolate D1) and Arthrobotrys bronchophaga (Isolate AB) had the smallest percentages of reduction among the tested isolates and showed the lowest predacious activity. The Isolates CG768, I31 and A183 were considered potential biological control agents of Ancylostoma spp. dog free-living stages, being directly influenced by the fungal inoculum concentration.

Scanning electron microscopy of Ancylostoma spp. dog infective larvae captured and destroyed by the nematophagous fungus Duddingtonia flagrans.

The interaction between the nematode-trapping fungus Duddingtonia flagrans (isolate CG768) against Ancylostoma spp. dog infective larvae (L(3)) was evaluated by means of scanning electron microscopy. Adhesive network trap formation was observed 6h after the beginning of the interaction, and the capture of Ancylostoma spp. L(3) was observed 8h after the inoculation these larvae on the cellulose membranes colonized by the fungus. Scanning electron micrographs were taken at 0, 12, 24, 36 and 48 h, where 0 is the time when Ancylostoma spp. L(3) was first captured by the fungus. Details of the capture structure formed by the fungus were described. Nematophagous Fungus Helper Bacteria (NHB) were found at interactions points between the D. flagrans and Ancylostoma spp. L(3). The cuticle penetration by the differentiated fungal hyphae with the exit of nematode internal contents was observed 36 h after the capture. Ancylostoma spp. L(3) were completely destroyed after 48 h of interaction with the fungus. The scanning electron microscopy technique was efficient on the study of this interaction, showing that the nematode-trapping fungus D. flagrans (isolate CG768) is a potential exterminator of Ancylostoma spp. L(3).

Variability of Meloidogyne exigua on Coffee in the Zona da Mata of Minas Gerais State, Brazil.

Minas Gerais is the major coffee-producing state of Brazil, with 28% of its production coming from the region of Zona da Mata. Four major species of root-knot nematode attacking coffee (Meloidogyne incognita, M. paranaensis, M. coffeicola, and M. exigua) have been reported from Brazil. To determine the variability in Meloidogyne spp. occurring in that region, 57 populations from 20 localities were evaluated for morphological, enzymatic, and physiological characteristics. According to the perineal pattern, all the populations were identified as M. exigua; however populations from the municipality of São João do Manhuaçu exhibited patterns very similar to M. arenaria. The identity of all the populations was confirmed by the phenotypes of esterase, malate dehydrogenase, superoxide dismutase, and glutamate-oxaloacetate transaminase. Thirteen populations (22.8%) showed the typical one-band (E1) esterase phenotype, whereas the others (77.2%) had a novel two-band phenotype (E2). No intraspecies variability was found in any population. All populations were able to reproduce on tomato, pepper, beans, cacao, and soybean. Reproduction was greater on tomato and pepper than on coffee seedlings, the susceptible standard.

Ditylenchus drepanocercus Rediscovered in the Neotropics Causing Angular Leaf Spots on Miconia calvescens.

During searches for pathogens to be used as classical biocontrol agents for Miconia calvescens (velvet tree), a devastating plant invader of Hawaii and French Polynesia, damaging angular leaf spots were found repeatedly. The etiological agent of this disease was identified as the nematode Ditylenchus drepanocercus. This nematode has a distinctive falciform appendage at the apex of the tail on both sexes, which allows easy identification. The nematodes were found in the lacunar parenchyma. Infected tissues have abnormally large cells (7 to 13 times the normal size). The lamina at infected areas is chlorotic, slightly thicker, and becomes necrotic with time. The best method of inoculation for this nematode was spraying plants with a suspension containing individuals of various stages on previously wounded leaves. Incubation period was determined to be 20 days.

Associação de biomateriais de implantes na reabilitação oral / Association of implants biomaterials in oral rehabilitation

Este presente trabalho reporta a avaliação clínica-radiográfica e a revisão de literatura pertinente sobre a utilização de diferentes materiais de implantes (hidroxiapatita - Biohapatita R; matriz óssea bovina liofilizada desmineralizada - OsseobondR; membrana de PTFE não reabsorvível - DentoflexR e implante de titânio 3iR Implant Innovation), na reabilitação do elemento dental 22 em uma paciente

Ultrastructure, Morphology, and Sporogenesis of Pasteuria penetrans.

ABSTRACT Pasteuria penetrans is a bacterial parasite of root-knot nematodes that shows great potential as a biocontrol agent. Scanning and transmission electron microscopy were used to study the ultrastructure, morphology, and sporogenesis of four isolates of P. penetrans. The effects of different Meloidogyne spp. and tobacco cultivars on sporangium size and morphology of endospores attached to the cuticle of second-stage juveniles (J2) of root-knot nematodes also were investigated. The P. penetrans isolates and their origins were P-20 from M. arenaria race 1 in Levy County, FL; P-100 from Meloidogyne sp. in Pasco County, FL; B-4 from Pratylenchus scribneri in Seminole County, FL; and P-120 from Meloidogyne spp. in Alachua County, FL. Sporangia of the four isolates were identical morphologically and similar in their dimensions, ranging from 2.39 to 3.42 mum in diameter and from 1.38 to 2.38 mum in height. Different Meloidogyne spp. and tobacco cultivars had no effect on sporangium size. Endospores attached to J2 were visualized in three forms: endospores retaining the sporangium wall, endospores covered with a thin exosporium, and endospores without covering. Sporogenesis of P. penetrans was similar to that of other gram-positive bacteria and generally matched the seven-stage scheme reported for Bacillus thuringiensis.

Temperature Effects on the Attachment of Pasteuria penetrans Endospores to Meloidogyne arenaria Race 1.

Pasteuria penetrans is a gram positive bacterium that prevents Meloidogyne spp. from reproducing and diminishes their ability to penetrate roots. The attachment of the endospores to the cuticle of the nematodes is the first step in the life cycle of the bacterium and is essential for its reproduction. As a preliminary study to a field solarization test, the effects of temperature on the attachment of P. penetrans on Meloidogyne arenaria race 1 were investigated. Preexposing second-stage juveniles (J2) of M. arenaria to approximately 30 degrees C in water before exposing them to endospores increased their receptivity to endospore attachment when compared to treating J2 at 25 degrees C or 35 degrees C. In tests with soil, highest attachment occurred when J2 were incubated in soil infested with endospores and maintained at 20 degrees C to 30 degrees C for 4 days. Heating J2 in soil to sublethal temperatures (35 degrees C to 40 degrees C) decreased endospore attachment. Incubating P. penetrans endospores in soil at 30 degrees C to 70 degrees C for 5 hours a day over 10 days resulted in reductions of endospore attachment to nematodes as temperatures of incubation increased to 50 degrees C and higher.

Informed consent with children and adolescents.

Nurses are becoming increasingly responsible for obtaining consent from children and adolescents for psychiatric treatment. The current practice of obtaining consent for participation in research is discussed. Consent from minors has implications for clinical treatment. This article examines power and powerlessness, and restrictiveness of the treatment setting in relation to nurses obtaining consent from children and adolescents.