Biochemical Changes in Freshwater Prawn Macrobrachium rosenbergii During Larval Development

Abstract.— Quantitative changes in the protein, lipid and carbohydrate were studied in the early larval stages of developing freshwater prawn Macrobrachium rosenbergii reared under fed and starved conditions to understand the relative importance of these nutrients in energy metabolism of the growing larvae. Larvae obtained from several females were stocked into three 250-L tanks at a density of 30 larvae/L. The feeding regime consisted of newly hatched Artemia nauplii. Protein was always the major organic constituent followed by lipids and then carbohydrates of both fed and starved larvae. Protein levels of both fed and starved larvae increased during development, suggesting an important role in morphogenesis. The decline of lipid during the larval growth that was more rapid for starved larvae, suggests a probable utilization of lipid as the major metabolic source of energy. Carbohydrates formed less than 5 and 2.4% of the larval dry weight of fed and starved larvae, respectively, suggesting their limited role in larval metabolism.     

Infectious disease serologic survey in free-ranging Venezuelan anacondas (Eunectes murinus).

Reptiles can harbor pathogenic microorganisms asymptomatically and serve as potential reservoirs of infection for humans, domestic animals, and other reptiles. Infectious diseases are also problematic for free-ranging reptile populations and are an important consideration in reptile reintroduction and translocation projects. There have been limited serologic studies of free-ranging reptiles for evidence of exposure to potential pathogens. In the present study, serum or plasma samples from five male and five female free-ranging Venezuelan anacondas (Eunectes murinus) were screened for antibodies to eastern, western, and Venezuelan equine encephalitis viruses, vesicular stomatitis virus, ophidian paramyxovirus, 19 Leptospira interrogans serovars, and Cryptosporidium serpentes. Antibodies to these agents were not detected, or antibody titers were low and possibly nonspecific. These results for the limited number of anacondas surveyed suggest that they do not serve as significant reservoirs for these infectious agents at this location.     

Impact of pig slurry on soil properties, water salinization, nitrate leaching and crop yield in a four-year experiment in Central Spain

Abstract. The repeated application of pig slurry to agricultural soils may result in an accumulation of salts and a risk of aquifer pollution due to nitrate leaching and salinization. Under Mediterranean conditions, a field experiment on a sandy loam soil (Typic Xerofluvent) was performed with maize (Zea mays) in 1998, 1999 and 2001 to study the effects of applying optimal (P1) and excessive rates (P3) of pig slurry on soil salinization, nitrate leaching and groundwater pollution. The rate of pig slurry was established considering the optimal N rate for maize in this soil (170, 162 and 176 kg N ha^?1 for 1998, 1999 and 2001, respectively). Pig slurry treatments were compared to an optimal N rate supplied as urea (U) and a control treatment without N fertilizer (P0). The composition of the slurries showed great variability between years. Mean NO₃^? leaching losses from 1998 to 2001 were 329, 215, 173 and 78 kg N ha^?1 for P3, P1, U and P0 treatments, respectively. The amount of total dissolved salts (TDS) added to the soil in slurry application between 1998 and 2001 was 2019 kg TDS ha^?1 for the P1 treatment and 6058 kg TDS ha^?1 for the P3 treatment. As a consequence, the electrical conductivity (EC) of the slurry‐treated soils was greater than that of the control soil. The EC correlated significantly with the sodium concentration of the soil solution. Over the entire experimental period, 2653, 2202 and 2110 kg Na ha^?1 entered the aquifer from the P3, P1 and P0 treatments, respectively. The P3 treatment did not significantly increase grain production in 1999 and 2001 compared with that achieved with the optimal N rate treatment (P1). This behaviour shows the importance of establishing application guidelines for pig slurry that will reduce the risk of soil and groundwater pollution.     

‘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