Light and electron microscopic observations of a segmented filamentous bacterium attached to the mucosa of the terminal ileum of pigs.

Segmented filamentous bacteria were seen attached to apical villous enterocytes of the terminal ileum in 15 of 2,766 live pigs submitted for necropsy over a 6-year period. Infected pigs ranged in age from 2 to 13 weeks. All pigs except 2, however, were greater than 4 weeks old and had been weaned. All infected pigs came from conventional commercial herds with intensive, all-indoor, confinement rearing management systems. The bacteria were gram negative or gram variable and were not associated with any clinical disease. Bacteria were more commonly attached to epithelium on the dome villi in the ileum. Electron microscopic examination revealed organisms composed of a cranial segment with a nipple-like appendage and several other segments making up elongated filaments of various lengths. Each filament was divided into segments by transverse septa. The nipple-like appendage served as an attachment apparatus by indenting the enterocyte surface without physically penetrating it. Occasionally, the colonizing bacterium was itself colonized by small rod-shaped bacteria that completely surrounded the filament.     

Bovine Cryptosporidiosis: Clinical and Pathological Findings in Forty-two Scouring Neonatal Calves

Cryptosporidia organisms were identified in 42 of 161 (26%) neonatal, diarrheic calves, over a 32 month period commencing July 1979. Forty of the 161 calves were submitted alive and cryptosporidiosis was diagnosed in 63% (25 of 40) of them. The cryptosporidia infected calves were usually one to two weeks old and came from 26 herds where the typical history was profuse, watery diarrhea in nearly all neonatal calves. The diarrhea usually started around one week of age, was unresponsive to all conventional antidiarrhea therapies, lasted for two or more weeks and was usually fatal. Twenty-nine (69%) of the cryptosporidia infected calves were submitted between December and February. These calves were often hutch reared.

Histopatholoical examination revealed large numbers of the coccidial parasite Cryptosporidium sp embedded in the microvilli of jejunal and ileal absorptive enterocytes of all affected calves. The organisms were identified as trophozoites and schizonts (asexual stages) and macrogametes (female sexual stages) with the electron microscope. Microgametes (male sexual stages) were not identified. Occasionally a merozoite (asexual stage) was also seen apparently burrowing into or about to be enveloped by a host microvillus. Observation of the organisms was much easier when diarrheic calves were submitted alive. Enterotoxigenic Escherichia coli were often cultured from intestines of dead calves and occasionally from calves submitted alive. Coronavirus particles were seen in one calf. In the last year of this study, oocysts were identified in fecal smears stained with May-Grünwald-Giemsa stain and fecal samples using a dichromate solution flotation technique.

     

‘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