The effect of dietary protein level on performance characteristics of coccidiosis vaccinated and nonvaccinated broilers following mixed-species Eimeria challenge.

A series of experiments were conducted to investigate the effect of starter diet protein levels on the performance of broilers vaccinated with a commercially available live oocyst coccidiosis vaccine before subsequent challenge with a mixed-species Eimeria challenge. Data indicated that an increasing protein concentration in the starter diet improved broiler performance during coccidiosis vaccination. Prechallenge performance data indicated that vaccination could decrease BW and increase feed conversion ratio. The time period most important for the observed effects appeared to be between 13 and 17 d of age. This reduction in performance parameters of vaccinated broilers compared with nonvaccinated broilers was eliminated by the conclusion of the experiments (27 d) in the diet groups with higher protein. Vaccination was effective at generating protective immunity against Eimeria challenge, as evidenced by increased (P < 0.05) BW gain, improved feed conversion, reduced postchallenge mortality, and reduced lesion development in vaccinated broilers compared with nonvaccinated broilers. These observations support numerous other reports that confirm live oocyst vaccination can be used effectively as a preventive against avian coccidiosis in commercially reared broilers. More important, these findings suggest that reduced protein concentration of starter diets can lead to significant losses in broiler performance when using a vaccination program to prevent coccidiosis.

Influence of orally administered CpG-ODNs on the humoral response to bovine serum albumin (BSA) in chickens.

Synthetic oligodeoxynucleotides containing CpG motifs (CpG-ODNs) have been reported to be effective mucosal adjuvants in mice when given orally. Studies on their effectiveness in chickens are currently very limited. This study investigated whether CpG-ODNs could adjuvant the immune response to BSA when given orally to a commercial line of SCWL chickens. In two experiments, performed over time, chickens were given selected concentrations of CpG-ODNs with BSA followed by 6 consecutive days of ad libitum access to drinking water containing 1.4 mg/ml BSA. Serum responses, and in some cases intestinal specific antibodies, were measured out to 33 days post-immunization. Birds receiving a single dose of CpG-ODN had consistently higher IgG, IgM, and IgA titers in the serum, dependent upon dose, and in specific areas of the intestine when compared to the non-immunized and BSA only groups. These findings suggest that a single oral CpG-ODN administration can accelerate the kinetics of antigen specific antibodies of all three isotypes in commercial-strain chickens immunized via the drinking water using common protein antigen.

Overexpression of human DNA topoisomerase II alpha by fusion to enhanced green fluorescent protein.

DNA topoisomerase (topo) II alpha is a major target for many anticancer agents. However, progress towards understanding how these agents interact with this enzyme in human cells and how resistance to these agents arises is greatly impeded by difficulties in expressing this gene. Here, we report on achieving a high level of expression of a full-length human topo II alpha gene in human cells. We started with the topo II alpha cDNA driven by a strong cytomegalovirus promoter and transiently transfected HeLa cells. Although topo II alpha mRNA was consistently detected in transfected cells, no exogenous topo II alpha protein was detected. By contrast, when the same cDNA was fused to an enhanced green fluorescent protein (EGFP), we detected a high level of expression at both mRNA and protein levels. The exogenous topo II alpha was localized to cell nuclei as expected, indicating that the fusion protein is properly folded. Furthermore, overexpression of the EGFP-topo II alpha fusion protein increased the sensitivity of the transfected cells to teniposide, suggesting that it functions as the endogenous counterpart. Thus, in addition to being used as a gene tag, the GFP fusion approach may be generally applicable for expressing genes, such as topo II alpha, that are difficult to express by conventional methods.

Participation of the intestinal epithelium and mast cells in local mucosal immune responses in commercial poultry.

The intestinal mucosa of commercial poultry is continually subjected to invasion or colonization by a wide array of potentially hostile enteric pathogens. Although, recent investigations have focused on lymphocyte involvement in immune responses in the intestine, lymphocyte-mediated immunity alone will not explain the barrier nature of mucosal membranes associated with rejection of many enteric pathogens upon secondary homologous challenge. Our laboratories have focused on nontraditional elements of mucosal immunity in poultry to better understand host-pathogen interactions in the intestine. Following classical and novel immunization procedures, we have identified an antigen-specific mechanism of immediate responsiveness of the mucosal epithelium characterized by epithelial chloride secretion. This mechanism, characteristic of intestinal anaphylaxis, is mediated by local immune elements. Similar mechanisms in mammals contribute to the barrier nature of mucosal membranes during pathogen challenge. To identify cells participating in these and similar responses, additional studies have described a role for mast cells in acute phase responses in the intestines of chickens experimentally challenged with Eimeria. To a more practical end, other experiments in our laboratories have characterized drinking water administration of BSA for elicitation of local and systemic antibody responses. These experiments have shown ad libitum drinking water administration of BSA to be as effective as i.p. administration of BSA; they present a novel approach to immunization of commercial poultry with protein vaccines. These investigations support continued research on host-pathogen interactions within the intestine of commercial poultry to better understand and control enteric pathogens through vaccination or immunomodulation.

Immunogenicity of ad libitum drinking water administration of bovine serum albumin in Leghorn chickens.

Oral administration of protein antigen in solution routinely leads to development of oral tolerance in most mammals but has been reported to be fully immunogenic in chickens. Previous studies, including several performed by our laboratory, have demonstrated that oral administration of discrete amounts of BSA for 6 consecutive days is fully immunogenic. This study was performed to determine immunoresponsiveness to protein antigen administered ad libitum at low levels in drinking water compared with i.p. and oral gavage routes of administration. Seven days following the last oral immunization, serum was assayed for IgG, bile for IgA, and tissue culture supernatant from 3 distinct lower intestinal regions for IgG and IgA in immunized and nonimmunized single-comb White Leghorn chickens. Systemic responses in the serum of experimental birds revealed a greater (P < 0.001) IgG response when BSA was administered via i.p. injection or by drinking water compared with gavage administration or nonimmunized controls. Responses measured in bile revealed that BSA administration in the drinking water resulted in a greater (P < 0.001) secretory IgA response compared with i.p. or gavage administration, and negative control groups. Intestinal antigen specific IgG, but not IgA, was elevated (P < 0.05) in all intestinal areas tested in birds immunized against BSA by drinking water and i.p. routes of administration, compared with other experimental groups. Taken together, the present experiments demonstrate that ad libitum drinking water administration of a protein antigen is as effective as i.p. administration or gavage routes of antigen exposure and potentially describe a novel approach to immunization of commercial poultry with purified protein antigens.