Light and scanning electron microscopy of the miracidium of Echinostoma paraensei (Trematoda, Echinostomatidae)

Echinostoma paraensei was described 1967 by Lie and Basch. Recently its natural definitive host, the aquatic rodent Nectomys squamipes, endemic in Brazil, was identified. As most of the echinostomatids, this species presents a zoonotic potential. The morphology and topography of the E. paraensei miracidium obtained from adult worms collected from the natural definitive host N. squamipes is described by light and electron microscopy. The arrangement and the dimensions of the epidermal plates are given. The eyespots are composed by two pairs of lenses measuring 6.03 microm. SEM observations shows that the miracidium body is covered by cilia except at the terebratorium region and the presence of alpha-tubulin in the cilia of the larvae is first recorded by immunelabeling. Nineteen papilla-like structures arranged in three axes and four groups were observed at the terebratoriun, this structure is retractable, presenting folds and a corrugated surface with profiles of cytoplasmic expansions, forming network of anatomizing folds.     

Proteomic survey of the pathogenic Mycoplasma hyopneumoniae strain 7448 and identification of novel post-translationally modified and antigenic proteins

Mycoplasma hyopneumoniae is an important pathogen for pigs, being the causative agent of enzootic pneumonia. Recently, the genome sequences of three strains, J, 7448 and 232 have been reported. Here, we describe the results of a proteomic analysis, based on two-dimensional gel electrophoresis of soluble protein extracts, immunoblot and mass spectrometry, which was carried out aiming the identification of gene products and antigenic proteins from the M. hyopneumoniae pathogenic strain 7448. A preliminary M. hyopneumoniae proteome map in two pH ranges (3-10 and 4-7) was produced. A total of 31 different coding DNA sequences (CDSs), including three hypothetical ones, were experimentally verified with the identification of the corresponding protein products by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. According to the Clusters of Orthologous Groups (COG) functional classification, the identified proteins were assigned to the groups of metabolism (13), cellular processes (5) and information and storage processing (4). Nine of the identified proteins were not classifiable by COG, including some related to cytoadherence and possibly involved in pathogenicity. Moreover, at least five highly antigenic proteins of M. hyopneumoniae were identified by immunoblots, including four novel ones (a heat shock protein 70, an elongation factor Tu, a pyruvate dehydrogenase E1-beta subunit and the P76 membrane protein). The now available proteome map is expected to serve as a reference for comparative analyses between M. hyopneumoniae pathogenic and non-pathogenic strains, and for methabolic studies based on cells cultured under modified conditions.     

Biochemical and histological responses of Rattus novergicus (Wistar) infected by Echinostoma paraensei (Trematoda: Echinostomatidae)

Tests were performed to evaluate the biochemical alterations in Rattus norvegicus after infection by the intestinal trematode Echinostoma paraensei. The rodents received 150 metacercariae each, serum samples were collected and the parasite load was quantified weekly until the fifth week of infection. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALKP), gamma-glutamyl transferase (GGT), bilirubin, glucose, total proteins and fractions and hepatic glycogen were determined. All the animals exposed to the metacercariae were infected in the first week and worms were recovered up to the third week after infection. The levels of AST, ALT, GGT, bilirubin and globulin rose in the first and/or second week and declined thereafter to levels near those of the control group. In contrast, the level of total proteins in the plasma fell significantly in the first week while the ALKP activity went down only in the fourth and fifth weeks in relation to the control group. There was no significant difference in the levels of albumin, glycogen and glucose. Infection by E. paraensei in R. norvegicus causes changes in the hepatic function, possibly resulting from the cholestasis produced by the partial obstruction of the bile duct by the helminths.     

Spatial distribution and sampling of Thaumetopoea pityocampa (Den. & Schiff.) (Lep. Thaumetopoeidea) populations on Pinus pinaster Ait. in Montesinho, N. Portugal

The pine processionary moth, Thaumetopoea pityocampa (Den. & Schiff.) is one of the most destructive pine pests in the Mediterranean countries. The first objective of this study was to analyse the spatial distribution of this insect on the tree on Pinus pinaster Ait., in northeast Portugal. The second objective was to develop two sampling plans: a sampling plan with fixed levels of precision for estimating T. pityocampa populations and a sequential sampling plan to classify the pest. Population estimates were made by registering the number of insect egg batches on 90 trees distributed over 3 stands during a 3-year study (1997–2000). Taylor's power law and Iwao regression were used to analyse the spatial distribution of the pest. Taylor's power law fitted the data better (r² = 0.775) than Iwao's regression model (r² = 0.704). The aggregation indices (b and β) were both significantly greater than one, indicating the aggregation of T. pityocampa egg batches. Optimal sample sizes for fixed precision levels of 0.10, 0.15 and 0.25 were estimated with Taylor's regression coefficients. Required sample sizes increased dramatically with increased levels of precision. The two sampling plans presented should be a tool for research on population dynamics and pest management decision.     

Evaluation of the Pollutant Content in Road Runoff First Flush Waters

Road runoff waters are able to convey a number of organic and inorganic pollutants originated by different non-point sources and by the road surface itself. Such pollutants can enter aquatic systems, thus contributing to water and soil contamination. This study was aimed at evaluating the pollutant content in first flush runoff waters from a highway located in Central Italy. A water sampling apparatus was devised able to collect only the first 10 L of runoff water. Water samples were analyzed by atomic absorption spectroscopy, ion chromatography, and gas chromatography-mass spectrometry in order to determine metals, ions, and polynuclear aromatic hydrocarbons, respectively. Soil samples were collected as well for metal and hydrocarbon determination. Results obtained in a field experiment carried out in two different seasons and in two different sites along the highway are reported. The study was extended to water samples collected in a strip of the same road not yet in use. Results obtained showed a temporal and a spatial variability due to the different period rainfall and to sites features, respectively. Within such a variability, a substantial agreement, in terms of order of magnitude, with other studies carried out in European sites was observed.     

Effect of extended aging of parma dry-cured ham on the content of oligopeptides and free amino acids

The effect of the dry-curing processing time on the release of oligopeptides and amino acids was evaluated with 158 Parma hams subdivided into three groups: (1) traditional processing (450 days); (2) extended processing (570 days); and (3) extended aging (690 days). Most of the oligopeptides and free amino acids detected increased up to the last deadline (690 days); a sharp increase of peptides below 400 Da was the main change in most aged hams. In particular, gamma-glutamyl dipeptides showed a remarkable increase during ham extended aging, acting like permanent taste-active compounds, being unsuitable for further enzymatic breakdown. The pH of fresh hams showed negative relationships (P < 0.001) with most peptides. With regard to free amino acids, the pattern was modified by different processing lengths, together with their taste categories, so that the amino acids having monosodium glutamate-like and bitter tastes were enhanced in more aged hams.     

Isolation and identification of two lipid transfer proteins in pomegranate (Punica granatum)

Lipid transfer proteins (LTPs) are a family of low molecular mass (7-9 kDa) polypeptides, the members of which share 35-95% sequence homology. These proteins are widely distributed throughout the plant kingdom and are receiving attention for their biochemical characteristics and biological activity. LTPs are indeed studied in different research fields varying from allergy to food technology, and numerous molecules belonging to this class are progressively being identified and investigated. Proteins from pomegranate juice were fractioned by cation exchange chromatography and analyzed by SDS-PAGE. Two proteins were identified as putative LTPs on the basis of their molecular weights and their electrophoretic behaviors under reducing and nonreducing conditions. Finally, proteins were purified and characterized by mass spectrometry. This analysis confirmed that the two polypeptides are LTPs on the basis of an amino acid sequence common to LTPs from other plant sources and cysteine content. The two proteins, named LTP1a and LTP1b, showed similar molecular masses but different immunological profiles when immunodetected with rabbit antibodies specific for Pru p 3 and human IgE from a patient suffering from pomegranate allergy. The demonstration of the existence of two immunologically unrelated LTPs in pomegranate confirms the variability and the complexity of the plant LTP family. This should be taken into account when the role of these proteins as elicitors of allergies to fruits is investigated and could help to explain the contradictory literature data on pomegranate allergy.     

Echinococcus ortleppi (G5) and Echinococcus granulosus sensu stricto (G1) loads in cattle from Southern Brazil

Echinococcus granulosus sensu stricto (G1) and Echinococcus ortleppi (G5) are haplotypes of the parasite formerly known as Echinococcus granulosus sensu lato, which in its larval stage causes cystic hydatid disease, endemic in Southern Brazil. Epidemiological and molecular knowledge about the haplotypes occurring in a region is essential to control the spread of the disease. The aim of this work was to analyze the haplotype frequency and fertility of hydatid cysts in cattle from the state of Rio Grande do Sul. Cysts were collected and classified according to their fertility status. DNA was extracted from protoscoleces and germinal layers and then used as template for the amplification of the cytochrome c oxidase subunit 1 gene by PCR. Amplicons were purified and sequenced, and the sequences were analyzed for haplotype identification. A total of 638 fertile cysts collected in the last ten years were genotyped. On average, G1 (56.6%) was more frequent than G5 (43.4%). In lungs, the G5 haplotype exhibited a higher parasite load (52.8%), whereas in the liver, G1 was more frequent (90.4%). The analysis revealed an increase in the frequency of G5 haplotype cysts during the period of sampling, and an increase in the abundance of fertile cysts has also been observed in the last several years. Most infertile cysts were genotyped as G1. The possible factors involved in the increase in the proportion of E. ortleppi (G5) and the consequences of this increase are discussed. This study suggests that the proportion of E. ortleppi (G5) loads in cattle may be increasing overtime.     

Anatomical characteristics, properties and use of traditionally used and lesser-known wood species from Mozambique: a literature review

Mozambique is a country with vast forestry resources that include native wood species with high commercial value. Thus, the trade of timber as raw material, as well as wooden finished products are commercial options of considerable valuable for the country. This work presents information about anatomical characteristics, physical and mechanical properties and use of some native wood species from Mozambique, namely, Afzelia quanzensis, Androstachys johnsonii, Erytrophleoum suaveolens, Khaya nyasica, Pterocarpus angolensis, Milletia stuhlmannii and the emerging lesser-known species Pericopsis angolensis, Sterculia appendiculata and Sterculia quinqueloba. The study concludes that these Mozambican wood species are similar in several aspects. They are generally described as very hard, dense, having high mechanical strength and durable. The study also concludes that although the lesser-known wood species, S. quinqueloba, has similar properties to the traditionally used one, A. quanzensis, it is less used because of its poorly known properties and potential uses. Thus, further research to determine the properties and suitable end uses of other lesser know species is likely to be beneficial to the country.