The effect of feeding a novel multistrain yeast fraction on European seabass (Dicentrachus labrax) intestinal health and growth performance

Fish were fed a single‐strain yeast fraction (SsYF; 2 g/kg) or a multistrain yeast fraction (MsYF; 0.8 g/kg) for 10 weeks. The results demonstrated significant (p ≤ 0.03) elevations in weight gain, specific growth rate, protein efficiency ratio, and feed conversion ratio in fish fed the yeast fraction‐supplemented diets. In the distal intestine, a significant elevation in microvilli density was observed after 5 and 10 weeks of dietary supplementation with MsYF and SsYF, respectively, compared to control fed fish (p < 0.001). A significant elevation (p = 0.02) in the perimeter ratio was observed in fish fed diets supplemented with the yeast fractions. After 10 weeks of feeding on the experimental diets, Rt‐qPCR demonstrated a significant downregulation (p < 0.05) in the stress response genes, heat‐shock protein 70 (hsp70) and proliferating cell nuclear antigen (pcna), in fish fed diets supplemented with the yeast fractions. Significant (p < 0.05) elevations in interleukin 1‐beta (il1β) and interleukin‐10 (il10) gene expression were observed in fish fed diets supplemented with the MsYF compared to the other dietary groups. These findings suggest that feeding an MsYF specifically at a lower incorporation rate < 1 g/kg, compared to a commercial SsYF at 2 g/kg, is effective in improving the intestinal health status and growth performance of European seabass.     

Capnographic documentation of nasoesophageal and nasogastric feeding tube placement in dogs

Objective: To evaluate the ability of capnography to document proper placement of nasoesophageal (NE) and nasogastric (NG) feeding tubes. This study was conducted in 3 phases. Phase I of this study was designed in order to test the efficacy of capnography to distinguish placement of a feeding tube in the alimentary tract versus the respiratory tract. Phase II was designed in order to document that carbon dioxide (CO₂) could be measured through a polyvinyl chloride (PVC) feeding tube. Phase III was performed in order to evaluate the technique of continuous monitoring during insertion of the feeding tube into the esophagus and stomach as would be performed during a clinical‐tube placement. Design: Prospective study. Setting: Research laboratory. Animals: 24 adult dogs. Interventions: In Phase I, sedated dogs were instrumented with an intratracheal catheter and an 8 French feeding tube placed nasally into the distal esophagus and later advanced into the stomach. In Phase II, dogs were anesthetized and an 8 French feeding tube was placed down the endotracheal tube, then into the esophagus and later advanced into the stomach. In Phase III, sedated dogs were instrumented with an 8 French feeding tube inserted intranasally and then advanced to the level of the nasopharynx, distal esophagus and, lastly, the stomach. Fluoroscopy was used in order to determine location of the feeding tube. Measurements and main results: Phase I measurements included respiratory rate and CO₂ from the trachea, esophagus, and stomach and pH of gastric fluid sample. Phase II measurements included respiratory rate and CO₂ from the endotracheal tube, feeding tube in the endotracheal tube, feeding tube in the distal esophagus, and feeding tube in the stomach. Phase III data collection included respiratory rate and CO₂ as the tube was passed through the nasal cavity, nasopharynx, esophagus and stomach. Phase I fluid samples were collected from 5 of the 9 dogs and had pH values from 1.68 to 4.20. In both phases, values for the respiratory rate and CO₂ from the esophagus and stomach were 0 ± 0, significantly lower (P < 0.001) than the values from the trachea. In Phase II, there was no significant difference between the respiratory rates (P = 0.886) and CO₂ (P = 0.705) readings obtained from the endotracheal tube compared to readings from the feeding tube in the endotracheal tube. In Phase III, there was a significant difference (P < 0.001) between the respiratory rates and CO₂ readings obtained from the nasal cavity and the nasopharynx when compared to those readings obtained from the esophagus and stomach. Measurement of CO₂ and respiratory rate resulted in a reading of 0 every time the feeding tube was in the esophagus or stomach. Conclusions: Capnography may be used in order to detect airway placement of NE and NG tubes.     

‘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.     

Culture of Differentiated Adult Rabbit Auricular Chondrocytes

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.     

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