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Biomedical subjects

H S Lillehoj

Publications and source records attributed to H S Lillehoj.

At least 19 recordsLinked to original sources
PubMed2007

Influence of Pediococcus-based probiotic on coccidiosis in broiler chickens.

Coccidiosis is the major parasitic disease of poultry and is caused by the apicomplexan parasites Eimeria. Drugs and live vaccines are the 2 main control measures of the disease; however, due to increasing concerns with prophylactic drug use and the high cost of vaccines, alternative control methods are needed. Recent evidence that various dietary and live microbial supplements can influence host immunity against enteric diseases prompted us to investigate the role of a Pediococcus-based probiotic on coccidiosis in broiler chickens. In the present study, we examined BW gains, oocyst shedding, and antibody responses of broilers fed the commercial probiotic MitoGrow. Day-old chicks were fed either a regular broiler diet or 1 of 2 probiotic diets supplemented with 0.1% (MG 0.1) or 0.2% MitoGrow. Chicks were orally challenged with 5,000 or 10,000 sporulated oocysts of Eimeria acervulina or with 5,000 Eimeria tenella oocysts on d 10 or 12 of age, respectively. In E. acervulina-infected birds, the MG 0.1 group improved (P < 0.05) weight gain as compared with the other 2 groups and reduced (P < 0.05) oocyst shedding in birds infected with 5,000 E. acervulina oocysts. In E. tenella-infected birds, Eimeria-specific antibody levels were higher (P < 0.05) in the Mito-Grow-fed groups, especially in the MG 0.1 birds, compared with the regular diet group, although their oocyst shedding and weight gains were not clearly improved. These results demonstrate that this Pediococcus acidilactici-based probiotic effectively enhances the resistance of birds and partially protects against the negative growth effects associated with coccidiosis, particularly when supplemented at 0.1% MitoGrow of the diet.

Animal Feed↗
PubMed2006

Monoclonal antibodies against chicken interleukin-6.

Monoclonal antibodies (mAb) were produced against a recombinant (r) chicken interleukin-6 (IL-6). Eight mAbs produced were tested for isotype; ability to inhibit recombinant forms of chicken (ch), human (h) and murine (m) IL-6; and recognition of rchIL-6 by Western immunoblotting. The mAb isotypes were represented by IgG1 (one), IgG2a (six) and IgG2b (one). In a mouse B9 hybridoma cell bioassay with rmIL-6, four mAbs effectively inhibited activity of rmIL-6. Further bioassays with the four mAbs at varying concentrations showed that two of these mAbs (1.20.7 and 1.26.4) were quite effective at inhibiting rmIL-6. Recombinant forms of ch, h and mIL-6 were all tested in a bioassay with the most potent inhibiting mAb (1.26.4), and this mAb was effective in inhibiting all three recombinant IL-6 proteins. Western immunoblotting revealed identification of the original IL-6 immunogen used for mAb production. Based upon inhibition of IL-6 activity in a standard bioassay and IL-6 recognition by Western immunoblotting, mAb 1.26.4 was judged the most useful antibody for future studies and applications.

Animals↗
PubMed2006

Fine-mapping of coccidia-resistant quantitative trait loci in chickens.

Two commercial, pure broiler lines with different susceptibility to coccidiosis were used to fine-map QTL associated with the previously identified marker LEI0101, located at 259 cM on chromosome 1 and shown to be significantly associated with disease resistance. Eight additional microsatellite markers linked to LEI0101 were used for genotyping of F(1) parents and F(2) offspring (n = 314), and their associations with oocyst shedding, as a marker of disease resistance, were determined in birds experimentally infected with Eimeria maxima. Single-point analysis of 4 families showed that logarithm of odds (LOD) scores at all marker loci were > 0.5, with the exception of marker LEI0071, located at 242 cM (LOD score = 2.45). Multipoint analysis showed a maximum LOD score between LEI0071 and LEI0101 at 254 cM (LOD score = 3.74). Although the LEI0071 marker was mapped near LEI0101 by linkage analysis, the physical location of LEI0071 was not identified. Further studies to determine the physical locations of these and other markers will allow additional application association mapping techniques using single nucleotide polymorphism markers.

Animals↗
PubMed2006

Immunopotentiating effect of a Fomitella fraxinea-derived lectin on chicken immunity and resistance to coccidiosis.

This study reports a novel immunopotentiating effect of a lectin (FFrL) extracted from the mushroom Fomitella fraxinea on poultry cell-mediated immunity and poultry coccidiosis. We describe the extraction of FFrL, its in vitro mitogenic activity and in vivo protection against an oral challenge infection with Eimeria acervulina. When tested on several cell types, crude FFrL agglutinated mouse erythrocytes and thymocytes and various other cells including murine and human cell lines. However, crude FFrL did not agglutinate human erythrocytes. Crude FFrL showed a potent mitogenic activity on chicken splenic lymphocytes, and at lower concentrations it exerted stronger mitogenic activity than Concanavalin A, a well-known potent mitogen for lymphocytes. Further, FFrL significantly induced (P < 0.05) nitric oxide secretion in HD11 cells and suppressed (P < 0.05) RP9 tumor cell growth in a dose-dependent fashion. When injected into 18-d-old chicken embryos followed by a posthatch oral E. acervulina challenge infection, FFrL treatment significantly protected chickens against weight loss associated with coccidiosis (P < 0.05). Injecting embryos with FFrL also resulted in significant reduction in oocyst shedding as compared with the control saline-injected birds (P < 0.05). The results of this study demonstrate that FFrL can be an effective growth promoting and immunostimulating agent in poultry during coccidiosis.

Agaricales↗
PubMed2005

Identification and characterization of the peroxiredoxin gene family in chickens.

Peroxiredoxin (PRX) is a crucial antioxidant protein that protects against endogenously produced peroxides in prokaryotes to eukaryotes. To date, 6 different isoforms have been identified in mammals. In this study, we describe the first members of the PRX protein family to be characterized in Chicken. Through bioinformatics analysis, we observed that at least 4 different classes of PRX protein have been evolutionarily conserved in chickens. Furthermore, in vitro functional assays of the candidate chicken PRX proteins demonstrated that they had levels of antioxidant activity similar to those of the mammalian enzymes. The expression patterns of the PRX transcript in several chicken tissues were not tissue specific, suggesting that they might play an essential role as a housekeeping gene in all tissues to protect against oxidative damage. In conclusion, the sequences of the putative members of this functional gene family in chickens could be effectively retrieved in silico through bioinformatics analysis, and the functionality of their gene products evaluated by in vitro comparative assay.

Animals↗
PubMed2005

Heat shock protein 90 genes of two species of poultry Eimeria: expression and evolutionary analysis.

Heat shock protein 90 (Hsp90) is 1 of the most abundant and evolutionarily conserved proteins. In most species, Hsp90 is essential for proper cell function. In this study, we present the molecular analysis of Hsp90 from Eimeria species, the causative agents of avian coccidiosis. The full-length Eimeria acervulina Hsp90 complementary DNA was isolated from intestinal intraepithelial lymphocytes of Eimeria-infected chickens. From evolutionary analysis and sequence identity, it is likely that Eimeria Hsp90 sequences described thus far encode the cytosolic versions of the protein. Although at the nucleotide and amino acid levels Eimeria tenella and E. acervulina Hsp90 are highly similar, their expression profiles differ considerably. Although E. tenella transcripts were detected in all developmental stages tested, E. acervulina transcripts were not found in oocysts undergoing sporulation or in fully sporulated oocysts, suggesting that messenger RNA expression may be regulated quite differently between Eimeria species.

Amino Acid Sequence↗
PubMed2004

Cell-mediated immune responses to a killed Salmonella enteritidis vaccine: lymphocyte proliferation, T-cell changes and interleukin-6 (IL-6), IL-1, IL-2, and IFN-gamma production.

Two experimental approaches were used to investigate the immunological responses of chickens to a commercial killed Salmonella enteritidis (SE) vaccine. In the first, the effects of host age on antigen-specific proliferative responses and cytokine production were examined. Compared with non-vaccinated controls, 4-wk-old vaccinated chickens showed higher proliferation to SE LPS and flagella. The lymphoproliferation responses to these antigens of 8-mo-old vaccinated chickens were not different compared to the non-vaccinated controls. Increased production of interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) by antigen-stimulated splenocytes following vaccination were, in general, more often observed in 4-wk-old compared with 8-mo-old chickens, whereas serum levels of these cytokines were consistently higher in the vaccinated birds compared with controls regardless of age. The second set of experiments were designed to determine the effects of SE vaccination on mitogen- or antigen-induced splenocyte proliferation and serum nitric oxide (NO) and cytokine levels. Splenocytes from vaccinated chickens stimulated with SE flagella showed significantly increased numbers of TCRgammadelta+ cells at 7 days post-vaccination compared with non-vaccinated birds. In contrast, no differences were noted with CD4+, CD8+, or TCRalphabeta+ cells at any time points examined. Higher levels of NO production were observed following stimulation with SE flagella at 4, 7, 11, and 14 days after SE vaccination while serum levels of IFN-gamma, IL-1, IL-6, and IL-8 were elevated only at day 7 post-vaccination. In conclusion, younger chickens mounted a more robust antigen-specific immune response to the SE vaccine compared with older birds and vaccination induced not only T-cell-mediated responses but also host innate and pro-inflammatory responses.

Age Factors↗
PubMed2004

Salmonella enteritidis clearance and immune responses in chickens following Salmonella vaccination and challenge.

Our previous work showed that the cell-mediated immunity (CMI) was enhanced by live Salmonella vaccine (LV). The objective of this study was to evaluate the impact of live and killed Salmonella vaccines on Salmonella enteritidis (SE) clearance and to determine if the clearance was mediated by cell-mediated and/or humoral immunity. Chickens were first immunized at 2 weeks of age followed by a booster dose at 4 weeks, challenged with live SE 2 weeks later (6-week-old) and tested for CMI, antibody response and SE clearance 1-week post SE-challenge (7-week-old). Spleen cell proliferation induced by SE-flagella and Concanavalin A (Con A) were significantly higher and SE shedding was significantly lower in the LV group. The splenic CD3 population was significantly lower and B cells were higher in the control group compared to all the SE-challenged groups (with and without vaccination). Serum antibody to SE-flagella and envelope were significantly higher in the KV group compared to all the other groups. These results suggest that LV protects against SE infection, probably by enhancing the CMI.

Animals↗
PubMed2004

Recent progress on the cytokine regulation of intestinal immune responses to Eimeria.

A variety of methods are available to combat avian diseases in the commercial setting, including improved farm management practices, use of antibiotic drugs, selection of disease resistant chicken strains, and manipulation of the chicken immune system. In the latter category, development of vaccines against the major avian diseases has become a priority for the poultry industry. With increasing demands for developing alternative control programs for many poultry diseases, it is important to understand the basic immunobiology of host-pathogen interactions in order to develop novel vaccination strategies. From studies carried out in many mammalian species, it is evident that host immune responses to intracellular pathogens are complex and involve many components of the host immune system. For enteric pathogens such as Eimeria and Salmonella, understanding cell-mediated immunity is most important because antibodies, although abundantly produced locally, can not access and act on these intracellular pathogens. In poultry, slow but increasing understanding of various components of host immune system mediating cellular immunity is opening new opportunities for thorough investigation of the role of thymus-derived lymphocyte subpopulations and cytokines in normal and disease states. This paper will review recent progress with chicken cytokines that have been characterized, and discuss various experimental strategies to enhance host immunity to pathogens using chicken cytokines.

Animals↗
PubMed2003

Analysis of cross-reactivity of five new chicken monoclonal antibodies which recognize the apical complex of Eimeria using confocal laser immunofluorescence assay.

For Apicomplexa (members) the host cell invasion is realized with the help of the organelles located at the apical tip of parasites. In this research paper the characterization of five chicken monoclonal antibodies (mabs) produced against Eimeria acervulina sporozoites is described. All mabs reacted with molecules belonging to the apical complex of chicken Eimeria sporozoites. On immunofluorescence assay (IFA) one mab, 8E-1, recognized an apical tip molecule present on all chicken Eimeria sporozoites, two mabs (8D-2 and HE-4) recognized an antigen present on the apical tip of the same two Eimeria species (E. acervulina and E. brunetti), another mab (5D-11) recognized an antigen present on the apical tip of other two species (E. acervulina and E. maxima) while one mab (8C-3) identified antigens present on the sporozoites and sporocysts wall of only E. acervulina. Besides the apical tip antigens, two mabs (HE-4 and 8D-2) recognized some proteins located in the anterior half of the sporozoites. Collectively, these mabs proved that the apical complex of chicken Eimeria sporozoites share one or more antigens that are expected to play a role in host cell recognition and invasion.

Animals↗
PubMed2003

Profiling local gene expression changes associated with Eimeria maxima and Eimeria acervulina using cDNA microarray.

Eimeria parasites show preferential sites of invasion in the avian intestine and produce a species-specific host immune response. Two economically important species, Eimeria acervulina and Eimeria maxima, preferentially invade and develop in the avian duodenum and jejunum/ileum, respectively. To investigate local host immune responses induced by parasite infection, global transcriptional changes in intestinal intraepithelial lymphocytes (IELs) induced by oral inoculation of chickens with E. acervulina or E. maxima were monitored using cDNA microarrays containing 400 unique chicken genes. Multiple gene transcripts were significantly up- or down-regulated following primary or secondary infection with E. acervulina or E. maxima. In general, infection by either parasite resulted in the expression changes of more genes following primary infection than following secondary infection, and E. acervulina caused more changes than did E. maxima. Although different regions of the small intestine were infected, similar changes in the levels of several cytokine mRNAs were observed in both Eimeria species following primary infection. Also identified was a set of transcripts whose expression was commonly enhanced or repressed in intestinal IELs of chickens infected with either parasite. Microarray analysis of chicken genes induced or repressed following Eimeria infection offers a powerful tool to enhance our understanding of host-parasite interactions leading to protective immunity.

Animals↗
PubMed2003

Increased lymphocyte subpopulations and macrophages in the ovaries and oviducts of laying hens infected with Salmonella enterica serovar Enteritidis.

Salmonella enterica serovar Enteritidis (SE) is a causative agent for human food poisoning cases throughout the world. The ovaries and the oviducts of the laying hens are the major sites of SE colonization from which vertical transmission to eggs occurs. In this study, Salmonella-induced changes in T lymphocytes, B lymphocytes and macrophages in the ovaries and oviducts were assessed after primary and secondary experimental inoculations of laying hen with SE. Statistically significant increases in the numbers of T cells (both CD4+ and CD8+) and macrophages were observed 7 to 14 days after primary inoculation, followed by a peak in B-cell numbers from the 14th day post-primary inoculation onwards in the secretory areas of the oviducts. The peak in lymphocyte numbers immediately preceded a decline in the rate of SE recovery from the reproductive tract beginning at day 14. The correlation of decreased Salmonella recovery with elevated lymphocyte and macrophage numbers strongly suggests that local cell-mediated immunity is involved in controlling SE injection in the ovaries and oviducts.

Animals↗
PubMed2003

Enhanced mucosal immunity against Eimeria acervulina in broilers fed a Lactobacillus-based probiotic.

The effect of feeding a Lactobacillus-based probiotic on intestinal intraepithelial lymphocyte (IEL) subpopulations and subsequent protection against coccidiosis was investigated in broiler chickens. Day-old male broilers were fed standard rations without control (CONT) or with a commercial probiotic (PROB) Primalac. Differences in IEL subpopulations were assessed by flow cytometry at 21 d postprobiotic treatment. At 25 d of age, a group of randomly selected birds from each diet was inoculated orally with 10,000 (per bird) sporulated oocysts of Eimeria acervulina and kept on the same diets. Fecal material, sera, and intestinal washes were collected 10 d postchallenge with E. acervulina. Birds on the PROB diet had more IEL expressing the surface markers CD3, CD4, CD8, and alphabetaTCR than those of the CONT diet. The probiotic-fed chickens produced less oocysts (P < 0.0001) compared to the untreated, control group (368 x 10(6) in CONT vs. 89 x 10(6) in PROB). The interferon-gamma levels in both serum and intestinal secretions were not significantly different between the two groups. However, CONT group showed higher antibody levels against a recombinant coccidial antigen in the intestinal secretions than the PROB group. No significant difference was found in serum antibody levels against the same antigen. These results dearly indicate that the probiotic bacteria impacted the local immune response as characterized by altered IEL subpopulations and increased the birds' resistance to E. acervulina as reflected by reduced oocyst shedding.

Animals↗
PubMed2003

Mapping quantitative trait loci associated with resistance to coccidiosis and growth.

To map QTL associated with disease resistance to avian coccidiosis and growth, two commercial broiler lines with different degrees of resistance to the disease were crossed to generate an F1 generation that was intercrossed to produce 314 F2 generation offspring. The F2 offspring were inoculated with sporulated oocysts of Eimeria maxima. Five disease-associated phenotypes were measured after the infection. Intertrait comparisons revealed that oocyst shedding was a good parameter for evaluating disease resistance or susceptibility. One hundred nineteen microsatellite markers, covering 80% of the chicken genome with an average marker interval of 25 cM, were used for genotyping of F1 parents and F2 offspring. Statistical analysis based on the data of four families revealed a locus on chromosome 1 associated with oocyst shedding (logarithm of odds = 3.46). The genetic mechanism of this locus appeared additive. The genomic scan also identified three potential growth QTL on chromosomes 1, 6, and 8. These results provide the foundation for further investigation to validate the QTL.

Animals↗
PubMed2002

Initiation of humoral immunity. I. The role of cytokines and hormones in the initiation of humoral immunity using T-independent and T-dependent antigens.

The early events during the initiation of immune responses following the injection of T-independent (lipopolysaccharide, LPS) and T-dependent (bovine serum albumin, BSA) antigens were studied in immature male chickens. Specifically, the role of cytokines and hormones in the initiation of humoral immunity against these antigens was investigated. Both interleukin-1 (IL-1) and tumor necrosis factor (TNF-alpha) increased significantly post-LPS but not post-BSA injection. While interleukin-2 (IL-2) significantly decreased post-LPS injection, IL-2 significantly increased post-BSA injection. Furthermore, corticosterone levels significantly increased and tri-iodothyronine (T(3)) levels significantly decreased post-LPS but not post-BSA injection. In this study, the results indicate that although LPS and BSA can induce a humoral antibody response in chickens, they activate different cytokines and neuroendocrine network systems.

Animals↗
PubMed2002

Initiation of humoral immunity. II. The effects of T-independent and T-dependent antigens on the distribution of lymphocyte populations.

The effect of injecting T-independent (lipopolysaccharide, LPS) and T-dependent (bovine serum albumin, BSA) antigens on the redistribution of lymphocyte populations in immature male chickens was investigated. In the blood, percentages of total T-cells (CD3+), T-helper cells (CD4+), and T-cytotoxic/suppressor cells (CD8+) significantly decreased post-LPS injection (PLI) but not post-BSA injection (PBI), while percentages of monocytes/thrombocytes (K1+) significantly increased PLI. Interleukin-1 receptor expression on blood lymphocytes increased significantly PLI and PBI. In the spleen, the percentages of total T-cells (CD3+) increased significantly PLI and PBI, macrophage (K1+) percentages increased significantly PLI, while B-cell percentages decreased significantly PLI. These results indicate that following antigen injection, there is a redistribution of peripheral blood lymphocytes (specifically T-lymphocytes) to secondary lymphoid organs and the kinetics and magnitude of the changes can differ according to the type of antigen used.

Animals↗
PubMed2002

Effect of vitamin A deficiency on host intestinal immune response to Eimeria acervulina in broiler chickens.

The effects of vitamin A (VitA) deficiency on the host intestinal immune response and disease susceptibility to coccidiosis were investigated in broiler chickens following oral infection with Eimeria acervulina (EA). Day-old male broilers were fed milo-soybean meal diets either with 8,000 IU VitA/kg feed (CONT) or without added VitA (A-DEF). At 25 d, a group of randomly selected birds from each treatment was inoculated orally with EA-sporulated oocysts. Intestinal immune response was assessed by the changes in the duodenum intraepithelial lymphocyte (IEL) subpopulations using flow cytometry at 35 d in in fected and noninfected birds. Concanavalin A (ConA)-induced spleen lymphocyte proliferation was tested using dimethylthiazol diphenyltetrazolium bromide colorimetric assay. Whether challenged or not with EA, A-DEF birds had fewer IEL expressing the surface markers CD3, CD4, CD8, alphabetaTCR, and gammabetaTCR. Without EA challenge, A-DEF birds had more surface IgA-expressing cells than CONT birds. Upon challenge, A-DEF chickens showed lower CD4+ IEL than CONT chickens. Following EA infection, CD8+ IEL increased in the CONT group, whereas no change was found in CD8+ IEL of A-DEF birds. A higher number of EA oocysts was recovered from A-DEF birds than from CONT birds (9.2 x 10(8) vs 5.4 x 10(8), respectively; P < or = 0.05). Serum samples taken 10 d post challenge showed higher antibody level against a recombinant coccidial antigen in A-DEF birds than in CONT birds. The A-DEF birds showed depressed ConA-induced lymphoproliferation response and produced lower serum interferon-gamma than CONT birds. These data show that VitA deficiency compromised local immune defenses of challenged birds, as reflected in lymphocyte profiles, oocyst shedding, and interferon-gamma levels in A-DEF birds.

Animals↗