Thesis (Ph. D.)--University of Wisconsin--Madison, 1958. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 87-90).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1949. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 51-52).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1955. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 58-60).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1944. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 63-67).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1965. / Typescript. eContent provider-neutral record in process. Description based on print version record.

Düsseldorf, Universiẗat, Diss., 2004.

An antibody library against Eimeria tenella sporozoites was constructed by phage display. Total RNA was isolated from the spleen, bone marrow, and ceca of immune chickens, and was used to reverse-transcribe cDNA. Heavy and light antibody variable genes were amplified from cDNA by the Polymerase Chain Reaction (PCR), using primer pairs that contain complementary sequences encoding a short linker sequence. The single-chain antibody fragment (scFv) was obtained by a secondary overlap PCR with primers that incorporate SfiI restriction sites, thus allowing for subsequent cloning into the phagemid vector pComb3X. Vector and scFv insert were digested with SfiI, ligated, and transformed into competent XL1-Blue Escherichia coli cells by electroporation, yielding a library with 7.4 x 107 total transformants. The culture was grown under carbenicillin selective pressure, rescued with helper phage, and the antibody-displaying phage was precipitated by PEG/NaCl, and subsequently used for panning. Five panning rounds were performed using cryopreserved E. tenella sporozoites, with a gradual increase of washing stringency to select for specific, highaffinity binders. A 1000-fold increase in phage output was obtained after 3 rounds of panning. There was clear enrichment of the positive clones over the panning rounds, with the 3rd round resulting in a 3,000-fold enrichment over the first one, as the binding clones became the dominant population in the library. Selected antibodies from the last round of panning were sequenced and characterized by immunoblotting. Soluble antibody fragments were produced in a non-suppressor E. coli strain, and recognized a 66-KDa sporozoite antigen on a Western blot. Primary cultures of chicken enterocytes were prepared in the hope of serving for invasion assays with E. tenella sporozoites. The isolation procedure, however, proved to be cumbersome and time-consuming. Future investigations will focus on purification and further characterization of antibodies selected from the constructed library. Such antibodies can be tested, alone or in combination, for their ability to block in vitro the invasion mechanism of E. tenella.

Abstract The purpose of this study is to investigate the probability of Eimeria tenella as a carrier for the transmission of chicken infectious anemia virus¡]CIAV¡^. One-day-old specific pathogen free¡]SPF¡^chickens were inoculated with CIAV, and then infected 20 days later with E. tenella. CIAV proteins were detected by immunofluorescent test in schizont, gametocytes and zygotes of E. tenella. Oocysts were purified from infected chicken by NaNO3 gradient centrifugation. Oocyst DNA was extracted by the treatments of SDS, proteinase K, DNase and chloroform. PCR template preparated from oocyst was used for amplification of CIAV DNA which is a 420 bp DNA fragment. The sequence of the PCR product from oocyst is similar to that of CIAV DNA from blood of infected chicken. The oocysts of E. tenella containing CIAV could also be suggested by inducing cytopathic effect after inoculated into cultured MSB1 cells. Our results indicated aprobability that E. tenella oocysts contain CIAV proteins and nucleic acids and thus E. tenella might paly an important role in the epizootiology.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1956. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 51-53).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1962. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 64-68).