Salmonella Enteritidis Vaccine Research Articles

Combination of Live and Inactivated Salmonella Vaccines to Protect Against Fowl Typhoid in Laying Hens.

Fowl typhoid (FT) caused by Salmonella Gallinarum (SG) is a poultry disease distributed worldwide that has been eradicated in commercial production of many developed countries but still persists in many developing countries. Vaccination is one of the main strategies to reduce mortality, clinical signs, and vertical or horizontal transmission. The aim of this work was to assess the protection against FT conferred by vaccines based on Salmonella Enteritidis (SE), SG, or a combination. Five experimental groups of birds, vaccinated with different live or inactivated SG and SE vaccines were included in the trial: 1) two doses of a SG-SE bivalent inactivated vaccine; 2) four doses of the live attenuated SE vaccine; 3) three doses of the live attenuated SE vaccine and two doses of the SG-SE bivalent inactivated vaccine; 4) two doses of the live attenuated SG9R vaccine; and 5) unvaccinated birds. At 28 wk of age, all hens were challenged with a virulent strain of SG, and mortality was recorded during the subsequent 15 days. The results showed that the plan that included only the inactivated vaccine did not show significant protection (P = 1), while the plan based on the administration of the attenuated strain of SE significantly reduced mortality in the group of birds (P = 0.0309). However, the highest levels of protection were obtained in the group of hens immunized with the combination of the inactivated vaccine and the live attenuated SE strain (P < 0.0001), which was statistically similar to the homologous protection conferred by the SG 9R strain, a vaccine used in many countries to control FT. These results demonstrate that the combination of existing vaccines together with strict biosecurity measures on farms may help improve the control of the pathogen in countries where FT in an emerging or reemerging disease.

Chitosan-nanoparticle-based oral Salmonella enteritidis subunit vaccine elicits cross-protection against Salmonella typhimurium in broilers

Non-typhoidal Salmonella infection is a significant health and economic burden in poultry industry. Developing an oral vaccine to induce robust mucosal immunity in the intestines of birds, especially cross protection against different Salmonella serotypes is challenging. Therefore, a potent oral vaccine platform that can mitigate different serotypes of Salmonella is warranted for the poultry industry. We reported earlier that the Salmonella enteritidis (SE) immunogenic outer membrane proteins (OMPs) and flagellin (FLA) entrapped in mannose chitosan nanoparticles (OMPs-FLA-mCS NPs) administered prime-boost (d-3 and 3-wk later) by oral inoculation elicits mucosal immunity and reduces challenge SE colonization by over 1 log10 CFU in birds. In this study, we sought to evaluate whether the SE antigens containing OMPs-FLA-mCS NPs vaccine induces cross-protection against Salmonella typhimurium (ST) in broilers. Our data indicated that the OMPs-FLA-mCS NPs vaccine induced higher cross-protective antibody responses compared to commercial Poulvac ST vaccine (contains a modified-live ST bacterium). Particularly, OMPs-FLA-mCS-NP vaccine elicited OMPs and FLA antigens specific increased production of secretory IgA and IgY antibodies in samples collected at both post-vaccination and post-challenge timepoints compared to commercial vaccine group. Notably, the vaccine reduced the challenge ST bacterial load by 0.8 log10 CFU in the cecal content, which was comparable to the outcome of Poulvac ST vaccination. In conclusion, our data suggested that orally administered OMPs-FLA-mCS-NP SE vaccine elicited cross protective mucosal immune responses against ST colonization in broilers. Thus, this candidate vaccine could be a viable option replacing the existing both live and killed Salmonella vaccines for birds.

Effects of Salmonella enteritidis Vaccination on the Expression of Innate Immune Molecules and Histone Modifications in the Follicular Theca of Laying Hens.

The aim of this study was to examine whether Salmonella enteritidis (SE) vaccination affects innate immune function and histone modifications responsible for epigenetic reprogramming in the follicular theca of laying hens. White Leghorn laying hens were administered the SE vaccine or phosphate buffered saline (PBS; control) one week before sample collection. The largest follicles (F1) were collected for total RNA and histone protein extraction. Gene expression levels of immune molecules (Toll-like receptors [TLRs], cytokines, and avian β-defensins [AvBDs]), and histone modifications in the follicular thecal tissues, were examined using real-time PCR and western blot, respectively. The results showed that the expression levels of TLR1-1, 2-1, 4, and 15 were upregulated by SE vaccination. Although vaccination caused no significant change in cytokine expression, AvBDl, 2, 4, and 7 expression levels were significantly upregulated in the vaccinated group. In addition, the relative density of histone H3-lysine9 dimethylation (H3K9me2) was increased by the vaccination. These results suggest that SE vaccination enhances innate immune functions in the ovary of laying hens, including upregulating TLR and AvBD expression, and is also associated with an increase in histone H3K9me2 in thecal cells.

Necrotizing and haemorrhagic hepatitis and enteritis in commercial layer pullets

ABSTRACTThis case report describes an episode of recurring severe necrotizing and haemorrhagic hepatitis and enteritis experienced in a flock of commercial layer pullets at 12 weeks of age and again at 18 weeks of age in Indiana. Pullets had been vaccinated at 10 weeks old using a trivalent Salmonella Enteritidis (SE)/Newcastle disease/infectious bronchitis oil-emulsion-inactivated vaccine. The pullets were found dead at 12 weeks with firm but friable, enlarged, haemorrhagic livers, enlarged spleens, and necrohaemorrhagic intestines. Histopathologic findings were consistent with a necrotizing and haemorrhagic enteritis and hepatitis. Livers had multiple intra-sinusoidal thrombi, intestines contained Gram-positive bacterial colonies, and spleens had marked lymphoid depletion. The pullets seemed to improve after antibiotic treatment. Pullets were vaccinated with an inactivated SE vaccine at 14 weeks of age. A second spike of mortality occurred at 18 weeks of age. Although clostridial enteritis and hepatitis were highly suspected in the two cases based on macroscopic and microscopic findings, no significant bacterial or viral agents were isolated from the livers and intestines. In summary, lesions in the liver and intestines are speculated to be due to repetitive vaccination, leading to an anamnestic response by the immune system, and resulting in an immune-mediated response. However, much of the pathogenesis is still unclear, and other causes such as unidentified infectious aetiology, transmissible amyloidosis, and hypersensitivity may need further investigation.

Combination of competitive exclusion and immunisation with a live Salmonella vaccine in newly hatched chickens: Immunological and microbiological effects

In addition to evaluating the efficacy potential of a combined use of vaccination and competitive exclusion (CE) against Salmonella exposure in chicks at 3-days of age, a live Salmonella Enteritidis vaccine (SE-LV) and a CE culture were tested for their ability to induce parameters of the innate immunity.Whereas the invasive SE-LV induced an influx of granulocytes and macrophages as well as an increased transcription of several cytokines in the caecal mucosa, the CE culture did not demonstrate any differences in these parameters compared to controls. It is therefore highly probable that the effects observed with CE cultures are not due to the rapid stimulation of the immune system. The combined use of both preparations did not result in an additive intestinal exclusion effect of the challenge strain more pronounced than that after single administration of the CE culture. The combined use of the Salmonella live vaccine and the CE culture resulted in an additive protective effect and prevented completely the systemic dissemination of the Salmonella challenge strain. To exploit the potential of combined use of CE and vaccination further and most effectively, live Salmonella vaccines are needed that are despite their attenuation in virulence still capable to induce both intestinal colonisation- and invasion-inhibition effects against Salmonella exposure.

Construction of a recombinant-attenuated Salmonella Enteritidis strain secreting Escherichia coli heat-labile enterotoxin B subunit protein and its immunogenicity and protection efficacy against salmonellosis in chickens

A live attenuated Salmonella Enteritidis (SE) strain secreting Escherichia coli heat-labile enterotoxin B subunit (LTB) protein was constructed as a new vaccine candidate. The comparative effect of this vaccine candidate was evaluated with a previously reported SE vaccine, JOL919. An asd+, p15A ori plasmid containing eltB-encoding LTB was introduced into a ΔlonΔcpxRΔasd SE strain, and designated as JOL1364. In a single immunization experiment, group A chickens were orally inoculated with phosphate-buffered saline as a control, group B chickens were orally immunized with JOL919, and group C chickens were orally immunized with JOL1364. The immunized groups B and C showed significantly higher systemic, mucosal and cellular immune responses as compared to those of the control group. In addition, the immunized group C showed significantly higher mucosal and cellular immune responses as compared to those of the immunized group B at the 1st week post-immunization. In the examination of protection efficacy, the immunized groups B and C showed lower gross lesion scores in the liver and spleen, and lower bacterial counts of SE challenge strain in the liver, spleen, and caeca as compared to those of the control group. The number of SE-positive birds was significantly lower in the immunized group C as compared to that of the control group at the 14th day post-challenge. In addition, the number of birds carrying the challenge strain in the caeca was significantly lower in the immunized group C than those in the immunized group B and control group at the 7th and 14th day post-challenge. These results indicate that immunization with the JOL1364 vaccine candidate can induce higher mucosal and cellular immune responses than those of the JOL919 for efficient protection against salmonellosis.

Enhanced protective immune responses against Salmonella Enteritidis infection by Salmonella secreting an Escherichia coli heat-labile enterotoxin B subunit protein

Escherichia coli heat-labile enterotoxin B subunit (LTB) protein is a potent mucosal adjuvant. In this study, the effect of an attenuated Salmonella secreting LTB protein as an adjuvant strain (JOL1228) for a live Salmonella Enteritidis (SE) vaccine candidate (JOL919) was evaluated. In a single immunization experiment, chickens immunized with a mixture of JOL919 (5 parts) and JOL1228 (1 part) showed enhanced mucosal and cellular immune responses and efficient protection against salmonellosis as compared to those unimmunized control chickens. In further analysis, chickens were primed at one day of age and were boosted at the fifth week of age to prolong immune responses and to maximize the protection efficacy against salmonellosis. The immunized groups B (prime and booster with JOL919), C (prime with JOL919-JOL1228 mixture and booster with JOL919), and D (prime and booster with JOL919-JOL1228 mixture) showed significantly higher humoral and cellular immune responses as compared to those in the unimmunized control group A. In addition, immunized groups C and D showed fewer gross lesions in the liver and spleen and a lower number of SE-positive organs, with the lowest bacterial counts in the SE challenge strain as compared to the control group. These results indicate that SE vaccination with the LTB strain can have an adjuvant effect on the vaccine candidate by enhancing immune responses, and that a prime-boost strategy with the addition of the adjuvant strain can efficiently protect birds against salmonellosis.

Genetic differ in TLR4 gene polymorphisms and expression involved in Salmonella natural and artificial infection respectively in Chinese native chicken breeds

Salmonella enteritidis (SE) is a foodborne pathogen that negatively affects both animal and human health. Genetic variations in response to pathogenic SE colonization or to SE vaccination were measured in chicken resource populations. Toll-like receptor 4 (TLR4) is part of a group of evolutionarily conserved pattern recognition receptors involved in the activation of the immune system in response to various pathogens and in the innate defense against infection. In this study, TLR4 was investigated the association of TLR4 gene polymorphisms with Salmonella natural infection situation of birds from two distinct Chinese genetic breeds. One SNP G1894C in the second intron of chicken TLR4 (chTLR4) was scanned in the two hens breed, which showed significant association with Salmonella natural infection situation (P<0.05). Genetic variations in response to pathogenic SE colonization also existed in distinct Chinese chicken resource population. In this study, mRNA expression of TLR4 was detected to investigate the association with the effect of artificial SE challenge in heterophil granulocytes and spleen of chicks from two distinct Chinese genetic breeds at 1, 3 and 10 day post-infection during the acute infection period. It clearly showed that young chicks' response to SE infection was regulated by TLR4 mRNA expression. The results suggest that genetics, time, gender, and interactions among these factors, play important roles in TLR4 mRNA basic values and copies modulation of SE mediated immune response in distinct Chinese chickens.

Henhouse Feeding Style and &lt;i&gt;Salmonella&lt;/i&gt; Enteritidis Contamination in Unvaccinated Flocks of Egg Farms, April 1994-March 2001

From April 1994 to March 2001, monitoring tests (surveillance) for contamination of Salmonella Enteritidis (SE) at commercial egg farms and farm traceback inspections for cases of SE foodborne illness were carried out in the Himeji Livestock Hygiene Service Center jurisdiction of Hyogo Prefecture, Japan. SE vaccination was not performed before the outbreak of SE-associated foodborne illness, and SE contamination of a farm was recognized. In the surveillance, the year average percentages of identified SE-positive farms were 6.1% in farms with floor feeding in an open-type henhouse (F-OH) and 12.7% in farms with cage feeding in a windowless-type henhouse (C-WH), but 0% in farms with only cage feeding in an open-type henhouse (C-OH). The highest identified proportion of SE-positive farms among all styles of farms was found (4.3%) in April 1997-March 1999. Farm traceback inspections were performed in 2 of 7 farms feeding by F-OH and in 5 of 9 farms feeding by C-WH in this period. Easier contamination with SE was found for C-WH or F-OH than for C-OH. Ninety percent of the birds (3,632,000 birds at 70 farms) are fed at these easier contamination farms by C-WH (89% of birds in 13% of farms) or F-OH (1% of birds at 10% of farms). Integrated sanitary requirements and SE vaccinations are especially necessary on farms feeding by F-OH or C-WH. Since 1999, these countermeasures have been performed, and SE foodborne illnesses and affected patients in Hyogo Prefecture have gradually decreased.

In vitro evaluation of live attenuated vaccines against Salmonella enteritidis: cell-mediated immune response

The use of Salmonella enteritidis (SE) live attenuated vaccines is one of the major tool to reduce this infection in commercial poultry. In this work, techniques, evaluating the presence and the expression of some cytokines, were studied to improve the knowledge of the cellular-mediated immune response following SE vaccination. This study demonstrated that SE vaccination enhances the production of INF-γ, IL-8, iNOs, while downregulates IL-1β. Between these immunologic parameters, the evaluation of INF-γ seems to be the most significant and easy test to plan and optimize SE vaccination programs.

In vitro evaluation of live attenuated vaccines against Salmonella enteritidis: humoral immune response

Salmonella enteritidis (SE) vaccination is one of the major tool to reduce the infection in commercial poultry. In this work, techniques, evaluating the presence of Ig-G and Ig-A in serum, tears and intestinal secretions, were studied to improve the knowledge of the humoral immune response following SE live attenuated vaccination. The Ig-G and Ig-A ELISA are important and easy tests to plan and optimize SE vaccination programs.

Comparison of Immunosuppression in Dry and Lactating Awassi Ewes due to Water Deprivation Stress

In seminomadic farming practice, dry and lactating ewes are exposed to different degrees of water deprivation, leading to stress followed by various disease outbreaks. This study compares quantitatively the immunosuppression to Salmonella Enteritidis (SE) fimbriae (14 and 21 kDa) and other major polypeptides (28.9, 37.7, 42.9, 68.0, 92.6 and 96.8 kDa) in water-deprived dry and lactating ewes. Sixteen dry and lactating multiparous Awassi ewes were divided into four treatment groups (A, A', B and B'). Ewes in groups A and B were lactating, whereas ewes in groups A' and B' were dry. All ewes were administered a killed SE vaccine, subcutaneously in the neck, at the initiation of the experiment. The water availability for ewes in groups B (lactating) and B' (dry) was ad libitum, while that for ewes in groups A (lactating) and A' (dry) was once every 4 days. A serum sample was collected from the jugular vein of each ewe at zero time (initiation of the experiment, when SE bacterin was delivered) and at 2, 9, 12, 15 and 18 days post SE vaccination. The percentage reduction in the level of humoral antibody response to polypeptides of > or = 21 kDa was more apparent in water-deprived lactating ewes of group A between 9 and 18 days post initiation of thirst. In this period, immunosuppression to polypeptides > or = 21 kDa was present in 14 out of 16 observations in group A (water-deprived lactating), with significant immunosuppression in 9 observations in relation to the respective control (p<0.05), while it was present in only 4 out of 16 observations in group A' (water-deprived dry), with significant immunosuppression in 2 observations (p <0.05). In conclusion, immunosuppression to polypeptides of > or =21 kDa is more significant in lactating water-deprived ewes in the period 9-18 days post initiation of thirst, a result that will influence our future sheep welfare awareness programmes targeting an elimination of the practice of water deprivation in seminomadic sheep farming.

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-γ 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-γ (IFN-γ) 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 TCRγδ + cells at 7 days post-vaccination compared with non-vaccinated birds. In contrast, no differences were noted with CD4 +, CD8 +, or TCRαβ + 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-γ, 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.

Antigen-specific lymphocyte proliferation and interleukin production in chickens immunized with killed Salmonella enteritidis vaccine or experimental subunit vaccines.

Lymphocyte proliferation and interleukin (IL)-2 and IL-6 levels in serum were measured as indicators of cell-mediated immunity after immunization of chickens with a commercial killed Salmonella enteritidis (SE) vaccine or experimental subunit vaccines of crude protein (CP) extract or the outer membrane protein (OMP). Significantly increased proliferative responses to SE flagella, but not lipopolysaccharide, porin, CP, or OMP, were observed at 1 wk postimmunizarion in the three vaccination groups. The responses to flagella were specific because flagella-induced proliferation was not seen in chickens immunized with adjuvant alone. Of the three immunization protocols, use of the killed SE vaccine appeared most effective because it induced higher flagella-stimulated lymphocyte proliferation at 1 and 2 wk postvaccination compared with the CP- and OMP-vaccinated groups. Significantly increased IL-2 and IL-6 levels in serum were seen at 1 wk postimmunization in the three vaccination groups compared with adjuvant alone, but there were no differences between the killed vaccine and the subunit vaccines at this time, and the levels of both lymphokines returned to baseline at 2 wk postimmunization. We conclude that cell-mediated immunity to SE after vaccination with the killed bacterial vaccine or subunit vaccines is transient and mainly limited to flagella.

Candidate Gene Approach: Potentional Association of Caspase‐1, Inhibitor of Apoptosis Protein‐1, and Prosaposin Gene Polymorphisms with Response to Salmonella enteritidis Challenge or Vaccination in Young Chicks#

Salmonella enteritidis (SE) contamination of poultry products is a major cause of food‐borne disease worldwide. Caspase‐1 and inhibitor of apoptosis protein‐1 (IAP‐1) were selected as candidate genes for chicken response to SE because their proteins play critical roles in the apoptotic pathway when intracellular bacteria interact with host cells. Prosaposin (PSAP) was selected as a positional candidate gene based on a previous quantitative trait loci (QTL) linkage study using the same population. The F1 offspring of outbred sires crossed with three diverse, highly inbred dam lines (two major histocompatibility complex‐congenic Leghorn lines named G‐B1 and G‐B2, and one Fayoumi line) were used to define the phenotypes. The F1 birds were involved in either pathogenic SE challenge, in which spleen and cecum content bacterial load were quantified, or SE vaccination, in which plasma antibody level to SE vaccine was evaluated. A polymerase chain reaction‐restriction fragment length polymorphisms (PCR‐RFLP) assay was developed to identify single‐nucleotide polymorphism (SNP) in the three genes. The F1 offspring of heterozygous sires for each gene were genotyped. The sire caspase‐1 gene was significantly associated with cecum content bacterial load (P = 0.04) in the three combined dam line crosses, and with spleen bacterial load in the G‐B1 cross (P = 0.02). The sire caspase‐1 gene was also significantly associated with antibody level to SE vaccine (P = 0.03) in F1 males in the three combined dam line crosses. The sire IAP‐1 gene was significantly associated with spleen bacterial load (P = 0.04) in the three combined dam‐line crosses, and interacted with dam‐line genetics (P = 0.01) for cecum content bacterial load. The sire PSAP gene significantly interacted with sex for spleen bacterial load (P = 0.004). This study is the first to demonstrate the association of SNPs for caspase‐1, IAP‐1, and PSAP genes with SE vaccine and with pathogen challenge response in chickens. #Journal paper No. 19835 of the Iowa Agriculture and Home Economics Experiment Station, Ames, IA 50011; Projects 2237 and 3521.

Natural resistance-associated macrophage protein 1 gene polymorphisms and response to vaccine against or challenge with Salmonella enteritidis in young chicks

Salmonella enteritidis (SE) contamination of poultry products is of global food-safety concern. The natural resistance-associated macrophage protein 1 (NRAMP1) affects host innate immunity to intracellular bacteria because of its ability to transport divalent cations in late endosome/lysosomes. Studying the association of the NRAMP1 gene and chicken innate immune response to SE can, therefore, aid understanding and enhancement of chicken genetic resistance to SE. The chicken NRAMP1 gene was investigated as a candidate gene for SE response in a unique resource population. Outbred broiler sires and three diverse, highly inbred dam lines (two major histocompatibility complex-congenic Leghorn and one Fayoumi line) produced F1 progeny that were evaluated as young chicks for either bacterial load in spleen and cecum after pathogenic SE inoculation or antibody level after SE vaccination. Thirty-seven single nucleotide polymorphisms (SNP) were identified in 3.1 kb of genomic DNA of the NRAMP1 gene. A PCR-RFLP assay was developed to identify a SNP in a conserved transport motif. The sire NRAMP1 gene SNP was associated (P < 0.02) with antibody level to SE vaccine for Sire 8170 offspring in the two Leghorn crosses. In Sire 8296 offspring, NRAMP1 was associated (P < 0.02) with spleen bacterial load in the combined dam-line crosses. This study demonstrated the association of a SNP polymorphism in a highly conserved region of NRAMP1 with SE vaccine and pathogen challenge response in young chicks, indicating that either NRAMP1 or a linked gene controls these SE-response traits.

Association of INOS, TRAIL, TGF-β2, TGF-β3, and IgL genes with response to Salmonella enteritidis in poultry

Several candidate genes were selected, based on their critical roles in the host's response to intracellular bacteria, to study the genetic control of the chicken response to Salmonella enteritidis (SE). The candidate genes were: inducible nitric oxide synthase (INOS), tumor necrosis factor related apoptosis inducing ligand (TRAIL), transforming growth factor β2 (TGF-β2), transforming growth factor β3 (TGF-β3), and immunoglobulin G light chain (IgL). Responses to pathogenic SE colonization or to SE vaccination were measured in the Iowa Salmonella response resource population (ISRRP). Outbred broiler sires and three diverse, highly inbred dam lines produced 508 F1 progeny, which were evaluated as young chicks for either bacterial load isolated from spleen or cecum contents after pathogenic SE inoculation, or the circulating antibody level after SE vaccination. Fragments of each gene were sequenced from the founder lines of the resource population to identify genomic sequence variation. Single nucleotide polymorphisms (SNP) were identified, then PCR-RFLP techniques were developed to genotype the F1 resource population. Linear mixed models were used for statistical analyses. Because the inbred dam lines always contributed one copy of the same allele, the heterozygous sire allele effects could be assessed in the F1 generation. Association analyses revealed significant effects of the sire allele of TRAIL-Sty I on the spleen (P < 0.07) and cecum (P < 0.0002) SE bacterial load. Significant effects (P < 0.04) were found on the cecum bacterial load for TGF-β3-Bsr I. Varied and moderate association was found for SE vaccine antibody response for all genes. This is the first reported study on the association of SNP in INOS, TRAIL, TGF-β2, TGF-β3, and IgL with the chicken response to SE. Identification of candidate genes to improve the immune response may be useful for marker-assisted selection to enhance disease resistance.

Association of MHC class I and class II gene polymorphisms with vaccine or challenge response to Salmonella enteritidis in young chicks.

Salmonella enteritidis (SE) is a worldwide source of salmonellosis in humans, caused mainly by consumption of contaminated poultry products. The major histocompatibility complex (MHC) class I and class II molecules, which function as antigen-presentation structures, are involved in both macrophage and dendritic cell immune response to Salmonella and may, therefore, play an important role in host resistance to infections. The chicken MHC class I and class II were investigated as candidate genes for immune response to SE. We characterized the complete MHC class I cDNA sequences for an outbred broiler line and four diverse highly inbred lines: two MHC-congenic Leghorn (G-B2 and G-B1), one Egyptian Fayoumi (M15.2), and one Spanish (Sp21.1), to define the allelic sequences within these lines, so that we might study the association between particular MHC polymorphisms and response to SE. The F1 offspring of outbred broiler sires crossed with three inbred lines (G-B1, G-B2, and Fayoumi) were evaluated as young chicks for either bacterial load in spleen and cecum after pathogenic SE inoculation or antibody level after SE vaccination. Alleles defined by a Lys(148)-->Met(148) polymorphism in the MHC class I alpha(2) domain were associated with spleen bacterial load after SE challenge. These results suggest that particular MHC haplotypes may contribute to control of responses to SE, and that particular polymorphisms may serve as markers for genetic resistance to SE in the chicken.

Candidate genes for resistance to Salmonella enteritidis colonization in chickens as detected in a novel genetic cross

Salmonellosis is a zoonotic disease that is problematic for both animal production and food safety. A novel genetic cross, named the Iowa Salmonella response resource population (ISRRP), was established to elucidate the genetic control of resistance to Salmonella enteritidis (SE) colonization in young chicks, to characterize unique resistance alleles, and to estimate gene interaction effects. Outbred broiler sires were mated with dams of diverse, highly inbred, light-bodied lines to produce an F 1 generation that was informative for all heterozygous alleles of the sires. Mating F 1 sires back to dams of the corresponding inbred line produced a backcross generation. To mimic the natural route of exposure and thus afford the opportunity to investigate mucosal immunity, pathogenic SE were inoculated into the esophagus of day-old chicks. After 1 week, the SE colonizing the cecal lumen and the spleen were enumerated. Candidate genes were selected for analysis based upon one of the two criteria. Functional candidates were genes with reported activity related to the tested traits. Positional candidates were genes mapped near microsatellites that were linked, in other phases of this project, with antibody levels to SE vaccine. Broiler sire alleles of the MHC class I, NRAMP1, PSAP, and IAP1 genes showed association with SE colonization in the F 1 generation of this novel disease resistance resource population.

Evaluation of 12 stabilizers in a developed attenuated Salmonella Enteritidis vaccine

The development of a stable live attenuated Salmonella Enteritidis (SE) vaccine, resisting heat stress during transportation and storage in unequipped tropical and subtropical zones of the world, is highly recommended. Twelve stabilizers were individually supplemented into a 9 ml volume of sterile distilled water resulting in concentrations of 1, 2, 3, 4 and 5%. A volume of 1 ml of attenuated live SE vaccine is added over the 9 ml of each concentration of the stabilizers. The differently stabilized SE vaccines were stressed at 55 °C for 48 h. The lowest percent reductions in SE cell viability by specified level of each stabilizer in ascending order were: 22.3% by 2% skim milk, 55.1% by 5% bovine serum albumin (BSA), 59.2% by 4% sorbitol, 74.4% by 3% maltose, 75% by 2% honey, 91.3% by 3% histidine, 96.9% by 1% heparin, 97.5% by 4% dextrose, 97.9% by 5% lactose, 99.4% by 5% sucrose, 99.5% by 2% gelatin, and 100% by 1–5% glycerol. In narrowing the concentration levels of skim milk to include 1.00, 1.25, 1.50, 1.75, 2.00, 2.25, 2.50, 2.75, and 3.00%, the 2.50% was the optimum level resulting in minimal percent reduction in SE cell viability of 18.9% after exposure to the defined heat stress.