Mycoplasma hyopneumoniae colonization of pigs sired by different boars

Differences in Mycoplasma hyopneumoniae colonization were evaluated in experimentally inoculated pigs sired by 3 different boars of the same genetic line. Forty-six pigs were used, including a treatment group and positive and negative control groups. The pigs were intratracheally inoculated with an M. hyopneumoniae suspension or with Friis media as a placebo. To evaluate differences in the magnitude of colonization during a 35-day period, nasal and tracheal swabs were collected weekly and tested by nested polymerase chain reaction (N-PCR). Temperature, weight and circulating antibodies were measured for 35 days. At 11 and 35 d postinoculation the pigs were necropsied and macroscopic and microscopic lesions were determined. A section of bronchus was tested by the indirect immunofluorescence test (IFAT), scanning electron microscopy (SEM) and N-PCR. The N-PCR results from the nasal and tracheal swabs showed that the pigs sired by one boar (B3) had a distinctive colonization pattern, different from that of the pigs from the other 2 boars and from the positive controls. SEM studies demonstrated that at 35 d postinoculation a higher proportion of B3 pigs had lower numbers of mycoplasmas attached to the cilia compared with B1 and B2 offspring. No significant differences were observed in temperature and weight gain among groups by ANOVA; however, with use of a 2 × 2 table, temperature differences were observed between pigs sired by boars B1 and B2 at 4 d postinoculation. No pigs seroconverted, showed gross or microscopic lesions, or had positive IFAT results. These results provide evidence of differences in patterns of colonization between pigs sired by different boars, suggesting a possible genetic effect.     

Landscape change under indirect effects of human use: the Savanna of Central Chile

The Chilean Intermediate Depression to the north of Santiago has experienced a physiognomical transformation from a Prosopis chilensis woodland to an Acacia caven savanna. Today P. chilensis trees are scarce and belong mostly to the larger size classes. By contrast A. caven seems to reproduce frequently and its populations consist of individuals of all size classes. In this paper we document these changes and report the results of tests aimed at determining the causes of these physiognomical changes. We found that livestock, leporids, introduced Mediterranean forbs and agriculture account for differences in seed dispersal and survival of A. caven and P. chilensis, which can explain the documented changes in the Chilean landscape.     

The Effect of Size and Diet on Gonad Production by the Chilean Sea Urchin Loxechinus albus

Conadal production was studied in Loxechinus albus (Molina, 1782) maintained in cages suspended from a long-line in the Estero Castro, Chiloé for 3 mo during the austral summer. The sea urchins were fed either an artificial diet or a natural diet consisting of the macroalgae Macrocystis pyrifera and Ulva sp. Both diets were tested for four size ranges: 40–45, 50–55, 60–65 and 70–75 mm diameter. For all four size ranges, highest gonad production was with the artificial diet. Gonad production was greatest in the 40–45 mm individuals with an increase of about 1,400% and 750% in the wet weight of gonads in individuals fed the artificial and natural diet, respectively. With the three other size ranges, the increase was nearly 100% with the artificial diet and nearly 0% with the natural diet The gonad index showed similar patterns, being highest in the smallest individuals. Small individuals fed the artificial diet would provide the most cost effective aquaculture as production is best. It is not necessary to grow L. albus to the minimal legal size for fisheries (70-mm diameter) for cost effective gonad production.     

Analysis of In vitro Fertilizing Capacity to Evaluate the Freezing Procedures of Boar Semen and to Predict the Subsequent Fertility

A porcine in vitro fertilization (IVF) system and seminal quality parameters of frozen–thawed boar semen were used to assess the effectiveness of two different thawing rates of frozen boar semen, and to address the question of whether differences between fertility of ejaculates could be predicted in a limited field trial. In the first experiment, two thawing procedures were analysed (37°C, 30 s; 50°C, 12 s) and no differences in sperm quality were found. However, when the procedure was 50°C, 12 s the IVF results showed a higher number of sperm per penetrated oocyte and a near 10 points higher rate of pronuclear formation. In the second experiment, the fertility results obtained in the limited field trial show to be efficient enough for application in a commercial use, especially for three of the employed boars (fertility ≥80%). In this limited study, the conventional seminal parameters are not accurate enough to discriminate good and bad boars in relation to fertility. On the contrary, parameters of in vitro penetrability are more precise to predict subsequent fertilities. As conclusion, the IVF fertilization system seems to be a good tool to evaluate the quality of frozen–thawed boar semen previous to its commercial way, to verify the bank semen storage quality and a good way to assay new sperm freezing procedures, as it is the more precise evaluating method in estimating the potential fertilizing ability.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001