Immunization of turkeys with Clostridium septicum alpha toxin-based recombinant subunit proteins can confer protection against experimental Clostridial dermatitis.

Clostridial dermatitis (CD), caused by Clostridium septicum, is an emerging disease of increasing economic importance in turkeys. Currently, there are no effective vaccines for CD control. Here, two non-toxic domains of C. septicum alpha toxin, namely ntATX-D1 and ntATX-D2, were identified, cloned, and expressed in Escherichia coli as recombinant subunit proteins to investigate their use as potential vaccine candidates. Experimental groups consisted of a Negative control (NCx) that did not receive C. septicum challenge, while the adjuvant-only Positive control (PCx), ntATX-D1 immunization (D1) and ntATX-D2 immunization (D2) groups received C. septicum challenge. Turkeys were immunized subcutaneously with 100 µg of protein at 7, 8 and 9 weeks of age along with an oil-in-water nano-emulsion adjuvant, followed by C. septicum challenge at 11 weeks of age. Results showed that while 46.2% of birds in the PCx group died post-challenge, the rate of mortality in D1- or D2-immunization groups was 13.3%. The gross and histopathological lesions in the skin, muscle and spleen showed that the disease severity was highest in PCx group, while the D2-immunized birds had significantly lower lesion scores when compared to PCx. Gene expression analysis revealed that PCx birds had significantly higher expression of pro-inflammatory cytokine genes in the skin, muscle and spleen than the NCx group, while the D2 group had significantly lower expression of these genes compared to PCx. Peripheral blood cellular analysis showed increased frequencies of activated CD4+ and/or CD8+ cells in the D1 and D2-immunized groups. Additionally, the immunized turkeys developed antigen-specific serum IgY antibodies. Collectively, these findings indicate that ntATX proteins, specifically the ntATX-D2 can be a promising vaccine candidate for protecting turkeys against CD and that the protection mechanisms may include downregulation of C. septicum-induced inflammation and increased CD4+ and CD8+ cellular activation.

Cellular and molecular insights on the regulation of innate immune responses to experimental aspergillosis in chicken and turkey poults.

Across the world, many commercial poultry flocks and captive birds are threatened by infection with Aspergillus fumigatus. Susceptibility to aspergillosis varies among birds; among galliform birds specifically, morbidity and mortality rates seem to be greater in turkeys than in chickens. Little is known regarding the features of avian immune responses after inhalation of Aspergillus conidia, and to date, scarce information on inflammatory responses during aspergillosis exists. Thus, in the present study, we aimed to improve our understanding of the interactions between A. fumigatus and economically relevant galliform birds in terms of local innate immune responses. Intra-tracheal aerosolization of A. fumigatus conidia in turkey and chicken poults led to more severe clinical signs and lung lesions in turkeys, but leukocyte recovery from lung lavages was higher in chickens at 1dpi only. Interestingly, only chicken CD8+ T lymphocyte proportions increased after infection. Furthermore, the lungs of infected chickens showed an early upregulation of pro-inflammatory cytokines, including IL-1ß, IFN-γ and IL-6, whereas in turkeys, most of these cytokines showed a downregulation or a delayed upregulation. These results confirmed the importance of an early pro-inflammatory response to ensure the development of an appropriate anti-fungal immunity to avoid Aspergillus dissemination in the respiratory tract. In conclusion, we show for the first time that differences in local innate immune responses between chickens and turkeys during aspergillosis may determine the outcome of the disease.

Aspergillus fumigatus infection may cause mortality in poultry, depending on species sensitivity. This study confirms the earlier activation of chickens' pro-inflammatory effectors to control Aspergillus dissemination, whereas turkeys' immune response enables the exacerbation of lung lesions.

Development of A recombinant nucleocapsid based indirect ELISA for the detection of antibodies to avian metapneumovirus subtypes, A, B, and C.

Nucleocapsid (N) protein is the most highly expressed of all avian metapneumovirus (aMPV) viral proteins and stimulates a substantial immune response in infected animals. Codon optimized recombinant N (rec-N) protein from aMPV subtypes A, B, and C were expressed using the baculoviral expression system in Trichoplusia ni (Tni) insect cells. A mixture of purified rec-N antigens from each subtype was used as a coating antigen and was evaluated in indirect ELISA (iELISA) to assess antibody response in serum samples collected from experimentally infected chickens and turkeys with different aMPV subtypes. Also, archived field serum samples that were collected from different poultry submissions were used. Receiver operating characteristic (ROC) analysis was performed using chicken and turkey serum samples that were confirmed by indirect fluorescent antibody (IFA) test for serostatus (positive n = 270, negative n = 610). The ROC analysis showed sensitivity and specificity of 97 % at a cut-off value of 0.25. The rec-N iELISA was compared with a commercial whole virus-based APV kit. The rec-N iELISA showed comparable results in detecting antibody response in aMPV infected chicken sera but was more sensitive in detecting early antibody response in aMPV infected turkey serum samples. Our results further confirm the presence of aMPV antibodies in Canadian domestic poultry populations. The developed aMPV-rec N iELISA offers a safe and valuable alternative to whole virus-based iELISA for serodiagnosis and seroepidemiological surveillance of the disease in domestic poultry.

Comparative investigation of IFN-γ-producing T cells in chickens and turkeys following vaccination and infection with the extracellular parasite Histomonas meleagridis.

The re-emerging disease histomonosis is caused by the protozoan parasite Histomonas meleagridis that affects chickens and turkeys. Previously, protection by vaccination with in vitro attenuated H. meleagridis has been demonstrated and an involvement of T cells, potentially by IFN-γ production, was hypothesized. However, comparative studies between chickens and turkeys on H. meleagridis-specific T cells were not conducted yet. This work investigated IFN-γ production within CD4+, CD8α+ and TCRγδ+ (chicken) or CD3ε+CD4-CD8α- (turkey) T cells of spleen and liver from vaccinated and/or infected birds using clonal cultures of a monoxenic H. meleagridis strain. In infected chickens, re-stimulated splenocytes showed a significant increase of IFN-γ+CD4+ T cells. Contrariwise, significant increments of IFN-γ-producing cells within all major T-cell subsets of the spleen and liver were found for vaccinated/infected turkeys. This indicates that the vaccine in turkeys causes more intense systemic immune responses whereas in chickens protection might be mainly driven by local immunity.

Effects of different levels of arginine and methionine in a high-lysine diet on the immune status, performance, and carcass traits of turkeys.

We postulated that the use of appropriate levels and proportions of arginine (Arg) and methionine (Met) in compound feed with high lysine content (Lys) would make it possible to fully exploit the growth potential of modern fattening turkey crossbreds, without compromising their immune system. The aim of this study was to determine the effect of different ratios of Arg and Met in diets with high Lys content on the performance and immune status of turkeys. The turkeys were assigned to 6 groups with 8 replicates per group and 18 birds per replicate. Six feeding programs, with 3 dietary Arg levels (90, 100, and 110%) and 2 dietary Met levels (30 and 45%) relative to dietary Lys content, were compared. During each of 4 feeding phases (weeks 0-4, 5-8, 9-12, and 13-16), birds were fed ad libitum isocaloric diets containing high level of Lys, approximately 1.83, 1.67, 1.49, and 1.20%, respectively. The dietary treatments had no effect on daily feed intake or body weight at any stage of the study. The protein content of the breast meat was higher in the treatments with the highest Arg level (110%) compared with the lowest Arg level (90%). Similarly, protein content was higher in the treatments with the higher Met level compared with the lower Met level. Higher plasma levels of tumor necrosis factor, interleukin 6 (IL-6), and immunoglobulin Y were found in turkeys fed diets with the lowest Arg content. An increase in Met content resulted in a decrease in plasma content of IL-6. In growing turkeys fed diets high in Lys, an Arg level of 90% relative to Lys can be used without negatively affecting production results and immune system. Regardless of dietary Arg levels, an increase in Met content does not stimulate the immune defense system and shows no effect on growth performance of turkeys in current trial.

Herpesvirus of turkey-vectored avian influenza vaccine offers cross-protection against antigenically drifted H5Nx highly pathogenic avian influenza virus strains.

Among the different vaccines used to control highly pathogenic avian influenza, an HVT vector-based live recombinant avian influenza vaccine, expressing the haemagglutinin gene of an H5N1 HPAI virus, has been used by the poultry industry since 2012. The objective of the study presented in this paper was to test the efficacy of the commercially available HVT-based recombinant H5 vaccine against antigenically drifted H5N1, H5N8 and H5N2 HPAI virus circulating in Egypt recently. Groups of SPF chicks vaccinated at day-old with the HVT-based recombinant H5 vaccine were challenged, along with non-vaccinated controls, with 106 EID50 each of H5N1, H5N2 or H5N8 HPAI virus at 28 days of age. The birds were monitored for clinical protection and virus shedding during a 10-day postchallenge period. Clinical protection levels were 90%, 90% and 80% following challenge with the H5N1, H5N2 and H5N8 field isolates, respectively. Challenge virus shedding was significantly reduced in vaccinated groups, with up to 40%, 30% and 20% of non-shedders, and 3.8, 3.3 and 2.8 log10 reduction in the amount of excreted virus following challenge with H5N1, H5N2 and H5N8 viruses, respectively. Analyses of the amino acid sequences of the HA proteins of challenge viruses and serological relatedness with the vaccine insert revealed significant antigenic divergences between the vaccine and the challenge viruses. These results provide further evidence of the potential of HVT-based recombinant H5 vaccine to provide cross-protection against antigenically drifted HPAI H5Nx viruses with strong control on virus shedding.

The effect of different dietary ratios of arginine, methionine, and lysine on the performance, carcass traits, and immune status of turkeys.

The research hypothesis postulated that the optimal dietary inclusion levels and ratios of lysine (Lys), arginine (Arg), and methionine (Met) can increase the growth potential of hybrid turkeys and limit metabolic disorders that weaken immune function. The experiment was carried out in a full rearing cycle, from 1 to 16 wk of age, in a two-factorial randomized design with 3 levels of Arg and 2 levels of Met (90, 100 and 110% of Arg, and 30 or 45% of Met, relative to the content of dietary Lys), with 6 groups of 8 replicates per group and 18 turkeys per replicate. In the first and second month of rearing, a significant dietary Arg-by-Met interaction was noted for daily feed intake and body weight gain, and a more beneficial effect was exerted by higher Met content and medium Arg content. Throughout the experiment, the higher dietary Met level increased the final body weight (BW) of turkeys (P = 0.001). Different dietary Arg levels had no influence on the growth performance of turkeys, but the lowest level decreased dressing yield (P = 0.001), and the highest level increased the percentage of breast muscles in the final BW of turkeys (P = 0.003). The lowest Arg level (90% of Lys content) undesirably increased the concentration of the proinflammatory cytokine IL-6 (P = 0.028) and decreased globulin concentration (P = 0.001) in the blood plasma of turkeys. The higher dietary Met level (45% of Lys content) increased plasma albumin concentration (P = 0.016). It can be concluded that higher dietary levels of Met (45 vs. 30% of Lys content) and Arg (100 and 110 vs. 90% of Lys content) have a more beneficial effect on the growth performance and immune status of turkeys.

The immune response of young turkeys to haemorrhagic enteritis virus infection at different levels and sources of methionine in the diet.

BACKGROUND: Haemorrhagic enteritis (HE) of turkeys was first described in 1937 in the USA, while in Poland it was first diagnosed in 1987. Polish haemorrhagic enteritis virus (HEV) isolates are usually low pathogenic and trigger a subclinical disease. Unfortunately, even the low- pathogenic HEV strains cause severe immunosuppression leading to secondary bacterial infections and huge economic losses. The objective of this study was to evaluate if the influence of Met on HEV infected turkeys immune response can be differentiated by both its level and source. Met is one of the amino acids that not only play a nutritional role but also participate in and regulate key metabolic pathways and immune response. In our study, the birds were assigned to 4 dietary treatments which differed in Met levels (0.55 and 0.78% in weeks 1-4 of age and 0.45 and 0.65% in weeks 5-8 of age, respectively) and sources (DL-methionine (DLM) or DL-methionine hydroxy analogue (MHA)). RESULTS: The HEV added the percentage of CD4+ cells and decreased the percentage of IgM+ cells in the blood, spleen and caecal tonsils (CTs) of turkeys. In addition, it increased the percentage of CD4+CD25+ cells in blood, and interleukin-6 (IL-6) level in plasma. The higher dose of Met led to a significant decrease in the percentages of CD4+, CD8+ and CD4+IL-6+ cell subpopulations in the blood of HEV-infected and uninfected turkeys and to an increase in the percentage of IgM+ B cells in CTs. Turkeys administered feeds with an increased Met content displayed a decrease in plasma IL-6 levels and an increase in plasma IgA levels. CONCLUSIONS: The results of this study indicate that HEV infection impairs the immune function in turkeys. Met content in the feed has a moderate effect on the immune response in HEV-infected turkeys. The source of this amino acid appears not be as important as its dose, because value of the analysed parameters did not differ significantly between turkeys receiving feeds with DLM or MHA. In the uninfected turkeys, the higher by 40% (than recommended by NRC) level of Met in the feeds had a positive effect on humoral immunity parameters.

Newcastle disease virus vectors expressing consensus sequence of the H7 HA protein protect broiler chickens and turkeys against highly pathogenic H7N8 virus.

Continuous outbreaks of highly pathogenic avian influenza (HPAI) viruses in commercial poultry have caused devastating losses to domestic poultry with a raising public health concern. The outbreaks of HPAI viruses have increased worldwide, including the North America. Therefore, vaccination has been considered as an alternative strategy for an efficient control of HPAI viruses. In this study, we aimed to generate Newcastle disease virus (NDV) vectored H7 serotype-specific vaccines by expressing the consensus sequence of the HA protein. Conventional NDV strain LaSota vector and a chimeric NDV vector containing the avian paramyxovirus type-2 F and HN protein were able to express the consensus sequence of HA protein. The protective efficacy of vaccines was evaluated in broiler chickens and in turkeys. One-day-old poults were prime immunized with the chimeric vector expressing the HA protein followed by boost immunization with LaSota vector expressing the HA protein or co-expressing the HA and NA proteins. Our vaccine candidates provided complete protection of broiler chickens from mortality and shedding of H7N8 HPAI challenge virus. Turkeys were better protected by boosting with the LaSota vector co-expressing the HA and NA proteins than the LaSota vector expressing only the HA protein. Our study demonstrated a potential use of heterologous prime and boost vaccination strategy to protect poultry against H7 HPAI viruses.

Effect of a post-hatch lipopolysaccharide challenge in Turkey poults and ducklings after a primary embryonic heat stress.

The effect of embryonic thermal manipulation on the post-hatch immune response to a lipopolysaccharide (LPS) challenge was studied in Pekin ducklings and turkey poults. Commercial duck and turkey eggs were distributed among four treatments: SS-Control (37.5 °C from embryonic day [ED] 1 to 25); SS-LPS (37.5 °C from ED1 to 25 + LPS at D0 [hatch]); HH-LPS (38 °C from ED1 to 25 + LPS at D0; SH-LPS (37.5 °C from ED1 to 10 and 38 °C from ED 11 to 25 + LPS at D0). At ED16 and ED24, the egg shell temperature of the duck and turkey eggs in the HH and SH treatments were higher (P ≤ 0.01) than the SS treatment. Ducklings and poults in the HH treatment had the lowest yolk free body weight at hatch (P ≤ 0.05). At 24, 48, and 72 h post-LPS injection, ducklings and poults in the HH-LPS treatment had significantly reduced BW compared with the SS-Con treatments (P ≤ 0.05). Ducklings and poults in the SH-LPS and HH-LPS treatments had increased plasma heat shock protein 70 (HSP70) and lower splenic HSP70 mRNA amounts than the SS-LPS treatments at 24, and 48 h post-challenge (P ≤ 0.05). At 48 and 72 h, macrophage nitric oxide (NO) production in ducklings and poults in the SH-LPS and HH-LPS treatments was lower than in the SS-LPS treatments (P ≤ 0.05). Ducklings and poults in the SH-LPS treatment had increased thymocyte proliferation compared to the SS-LPS treatment at 24, 48 and 72 h (P ≤ 0.05). At 24 h, ducklings in the SH-LPS treatment had increased splenic IL-10 and reduced IFNγ and IL-6 mRNA abundance. However, both ducklings and poults in the HH-LPS treatment had increased IFNγ, and IL-10 mRNA abundance compared to the SS-LPS treatment (P ≤ 0.05). At 48 h, SH-LPS ducklings and poults had lower splenic IL-10 mRNA abundance (P ≤ 0.05) while the HH-LPS treatment resulted in comparable splenic IL-10 mRNA compared to the SS-LPS treatment (P ≥ 0.05). Ducklings and poults in the SH-LPS treatment had increased thymic and splenic CD8+/CD4+ ratios at 24 h versus the SS-LPS treatment (P ≤ 0.05). In conclusion, embryonic thermal manipulation from ED11-25 increased extracellular HSP70 release, thymocyte proliferation and IL-10 but decreased splenic HSP70 and IFNγ mRNA amounts at 24 h post-LPS injection. This suggests that mild heat stress during the later stages of incubation could potentially prime the embryonic immune system thereby enhances the immune response as earlier than 24 h to eliminate the inflammatory response without affecting the growth performance by increase the extracellular release of HSP70 in both ducklings and poults. Continuous exposure to the small increase in temperature from ED 1-25 (HH) caused an imbalance between pro (IFNγ)- and anti-inflammatory cytokines(IL-10) which affects hatchling responses to an inflammatory challenge and increased mortality. The amount of extracellular HSP70 could potentially play an important role in modulating the immune response against inflammatory challenges.

Field evaluation of maternal antibody transfer from breeder turkey hens to egg yolks, egg whites, and poults.

Immunoglobulins, which are passed vertically from hens to their progeny, are first present in the eggs but with time also in the developing embryos and eventually in the serum of hatching chicks, and have protective function during embryogenesis and in the first few weeks of birds' life, before the immune system becomes fully efficient. Considering the above fact, the aim of this study was to determine total levels of IgM and IgY as well as specific IgY antibody titers against selected pathogens in the serum of breeder turkeys and their progeny, as well as in egg yolks and egg whites. Study results demonstrated that the level of IgY antibodies in the serum of turkey breeder hens reached 22.04 mg/mL on average in the whole egg laying cycle. In addition, the mean transfer percentage of IgY antibodies from turkey layers to their progeny reached approximately 31.4%, but the level of this transfer differed depending on pathogen character and accounted for 33.2%, 51.9%, 45.1%, and 44.3% in the case of antibodies against avian metapneumoviruses, Newcastle disease virus, Ornithobacterium rhinortacheale, and Pasteurella multocida, respectively. Antibody percentage transfer differed also as affected by the stage of the egg production cycle. Study results confirmed the earlier observed dependency concerning the class of antibodies transferred to eggs from laying hens, and while the IgY were mainly detected in the egg yolk extracts, the IgM were found only in egg white extracts; in comparison to IgY, the IgM antibodies were not transferred to the serum of turkey poults. To our best knowledge, this is the first study that describes in detail the phenomenon of maternal antibody transfer in turkeys.

Cross-protective Salmonella vaccine reduces cecal and splenic colonization of multidrug-resistant Salmonella enterica serovar Heidelberg.

Salmonella vaccine strategies for food-producing animals have typically focused on a specific serovar that either causes production losses due to morbidity/mortality or is an important food safety pathogen for a particular food commodity. The Salmonella enterica serovar Typhimurium BBS 866 vaccine strain was designed to reduce serovar specificity to provide cross-protection against diverse Salmonella serovars, thereby broadening its applicability for multiple animal and poultry species. We reported cross-protection of the BBS 866 vaccine in swine [Vaccine 34:1241-6]. In the current study, we extend the efficacy of the Salmonella vaccine to a poultry commodity by revealing significant reduction of multidrug-resistant Salmonella enterica serovar Heidelberg colonization of the cecum and systemic dissemination to the spleen in BBS 866-vaccinated turkeys. Transcriptional analysis of whole blood from BBS 866-vaccinated turkeys revealed down-regulation of metabolic and immune genes (KCNAB1, ACOD1, GPR17, ADOR2AB, and IL-17RD), suggesting limited leukocyte activation without an overt peripheral inflammatory response to vaccination.

Effect of 25-hydroxycholecalciferol supplementation on turkey performance and immune cell parameters in a coccidial infection model.

This study was conducted to identify the effects of 25-OH cholecalciferol supplementation to turkeys on the immune cells parameters, fecal coccidial oocyst shedding, macrophage nitric oxide production, T regulatory cell cytokine production, and production parameters during a coccidial challenge. A total of 200 1-day-old turkey poults were supplemented with 27.5, 55, 82.5, or 110 µg/kg 25-OH cholecalciferol and challenged or not challenged with coccidial oocysts in a 4 × 2 factorial set up of treatments. Birds fed 110 µg/kg of 25-OH cholecalciferol and infected with coccidial oocyst had 41% lower (P < 0.05) fecal oocyst and 53% higher (P < 0.05) macrophage nitric oxide production than the birds fed 27.5 µg/kg of 25-OH cholecalciferol and infected with coccidial oocyst at 5 d post-coccidial infection. Birds fed 82.5 µg/kg 25-OH cholecalciferol and infected with coccidial oocyst had 5-fold higher (P < 0.05) IL-1 mRNA amounts than the birds fed 27.5 µg/kg of 25-OH cholecalciferol and infected with coccidial oocyst. Birds fed 110 µg/kg 25-OH cholecalciferol and infected with coccidial oocyst had 5.3-fold higher (P < 0.05) IL-10 mRNA amounts than the birds fed 27.5 µg/kg of 25-OH cholecalciferol and infected with coccidial oocyst at 5 d post-coccidial infection. CD4+CD25+ cells from birds fed 110 µg of 25-OH cholecalciferol and infected with coccidial oocyst had 12-fold higher (P < 0.05) IL-10 mRNA than that from the birds fed 27.5 µg/kg of 25-OH cholecalciferol and infected with coccidial oocyst. In conclusion, supplementing birds with 101 µg/kg 25-OH cholicalciferol decreases coccidial oocyst shedding in the feces and could be a nutritional strategy to reduce the coccidial infection and spread in turkeys.

The effect of the source and dose of manganese on the performance, digestibility and distribution of selected minerals, redox, and immune status of turkeys.

The aim of this study was to determine the effect of various levels of manganese added to the diet of growing turkeys in the conventional form of MnO or in the form of NP-Mn2O3 nanoparticles on growth performance, absorption, and accumulation of Mn, Zn, and Cu, and antioxidant and immune status. The experiment was conducted on 1080 one-day-old Hybrid Converter turkeys randomly assigned to 6 groups with 10 replications, in a two-factor design with three dosages of manganese - 100, 50, and 10 mg/kg, and two sources-manganese oxide (MnO) and manganese nanoparticles (NP-Mn2O3). Neither reducing the addition of Mn from 100 to 50 or even 10 mg/kg of the diet nor replacing MnO with NP-Mn2O3 had a negative effect on the growth performance of the turkeys. Replacing MnO with NP-Mn2O3 in the turkey diet improved ileal digestibility of Mn and decreased accumulation of Cu in the liver and breast muscle. The study showed that irrespective of the form used, reducing the level of Mn supplementation of the diet from the 100 mg/kg recommended by British United Turkey to 50 or 10 mg/kg decreased its ileal digestibility and increased its accumulation in the liver, breast muscle, and skin. Reducing the addition of Mn to the turkey diet increased intestinal absorption of Zn and reduced accumulation of Zn and Cu in the liver, breast muscle, and skin. It did not increase oxidation processes in the liver or breast muscle of the turkeys. Reducing the addition of Mn to the turkey diet stimulated the immune system, which was manifested by stimulation of B cells to produce immunoglobulin M and by the release of the cytokine IL-6, but did not intensify apoptosis. The results of the study indicate that the recommended manganese supplement in turkey diets can be reduced. The use of manganese nanoparticles in turkey feeding requires further study.

The effect of manganese nanoparticles on apoptosis and on redox and immune status in the tissues of young turkeys.

The aim of the study was to determine whether the use of Mn nanoparticles would make it possible to reduce the level of this micronutrient added to turkey diets without adversely affecting growth performance, antioxidant and immune status, or apoptosis. The experiment was conducted on 6 groups of turkeys with 10 replications in a two-factor design with 3 dosages of manganese, 100, 50 and 10 mg/kg, and 2 sources, manganese oxide (MnO) and manganese nanoparticles (NP-Mn2O3). The study showed that irrespective of the form of Mn used, reducing the Mn level recommended by British United Turkeys for supplementation of the diet of young turkeys from 100 mg/kg to 10 mg/kg increases the content of this element in the blood with no adverse effect on growth performance or the immune system. The reduction in Mn supplementation in the form of NP-Mn2O3 from 100 to 50 and even to 10 mg/kg of turkey diet has no negative effect on antioxidant defence in young turkeys. A 50% reduction of the recommended Mn level in the form of MnO enhances lipid oxidation processes. Replacing MnO with NP-Mn2O3 in the turkey diet probably can increase apoptosis in young turkeys. On the other hand, irrespective of the form of Mn used, reducing supplementation of the turkey diet with this element from 100 to 50 and even to 10 mg/kg probably can reduce apoptosis.

Antibody titers in turkeys increase after multiple booster vaccinations with an attenuated Salmonella live vaccine.

OBJECTIVE: Human Salmonellosis is one of the most frequently reported foodborne zoonoses in the European Union. The most common source of human infections is the consumption of poultry products. Besides management and hygiene practices vaccination of poultry livestock is seen as one way to reduce Salmonella infections in humans. Turkey flocks in Europe are frequently infected with Salmonella and until recently there was no live vaccine for turkeys available. The aim of the present study was to examine the development of humoral antibodies after repeated vaccination with a bivalent live Salmonella vaccine containing attenuated Salmonella Typhimurium and Salmonella Enteritidis strains. Furthermore the colonization of the caecum with the vaccine strains and their spread to liver and spleen as well as the course of their fecal excretion was observed. RESULTS: Antibody production was hardly detectable after the first vaccination but increased after booster vaccinations. Both the Salmonella Enteritidis and the Salmonella Typhimurium vaccine strain were reisolated from caecum contents and organ samples. After booster vaccinations the re-isolation rates were reduced. The shedding of the vaccine strains was most pronounced after the first vaccination.

Allocation of Interferon Gamma mRNA Positive Cells in Caecum Hallmarks a Protective Trait Against Histomonosis.

Histomonosis is a parasitic disease of gallinaceous birds characterized by necrotic lesions in cacum and liver that usually turns fatal in turkeys while it is less severe in chickens. Vaccination using in vitro attenuated Histomonas meleagridis has been experimentally shown to confer protection against histomonosis. The protective mechanisms that underpin the vaccine-induced immune response are not resolved so far. Therefore, the actual study aimed to evaluate the location and quantitative distribution patterns of signature cytokines of type 1 [interferon gamma (IFN-γ)] or type 2 [interleukin (IL)-13] immune responses in vaccinated or infected hosts. An intergroup and interspecies difference in the spatial and temporal distribution patterns of cytokine mRNA positive cells was evident. Quantification of cells showed a significantly decreased percentage of IFN-γ mRNA positive cells at 4 days post-inoculation (DPI) in caeca of turkeys inoculated exclusively with the attenuated or the virulent inocula, compared to control birds. The decrement was followed by a surge of cells expressing mRNA for IFN-γ or IL-13, reaching a peak of increment at 10 DPI. By contrast, turkeys challenged following vaccination showed a slight increment of cecal IFN-γ mRNA positive cells at 4 DPI after which positive cell counts became comparable to control birds. The increase in infected birds was accompanied by an extensive distribution of positively stained cells up to the muscularis layer of cecal tissue whereas the vaccine group maintained an intact mucosal structure. In chickens, the level of changes of positive cells was generally lower compared to turkeys. However, control chickens were found with a higher percentage of IFN-γ mRNA positive cells in cecum compared to their turkey counterparts indicating a higher resistance to histomonosis, similar to the observation in immunized turkeys. In chickens, it could be shown that the changes of cytokine-positive cells were related to variations of mononuclear cells quantified by immunofluorescence. Furthermore, gene expression measured by reverse transcription quantitative real time PCR confirmed variations in organs between the different groups of both bird species. Overall, it can be concluded that a proportionally increased, yet controlled, allocation of IFN-γ mRNA positive cells in caeca hallmarks a protective trait against histomonosis.

The effect of raw and fermented rapeseed cake on the metabolic parameters, immune status, and intestinal morphology of turkeys.

This study evaluated the effects of including 15% of raw or fermented rapeseed cake (RRC and FRC, respectively) in turkey diets on the growth performance, antioxidant and immune status, and intestinal morphology of birds. Rapeseed cake was fermented using the commercial 6-phytase enzyme preparation, and dried. A total of 1,350 day-old female Hybrid Converter turkeys were randomly allocated to 3 dietary treatments with 9 replicates per treatment and 50 birds per replicate. All diets were isonitrogenous and isocaloric, and contained various protein sources. In the control group (C), soybean meal was the main source of dietary protein, and the remaining groups were fed diets containing 15% of RRC or FRC. Fermentation considerably reduced the concentrations of phytate-phosphorus and glucosinolates in rapeseed cake. In comparison with RRC, turkeys receiving FRC achieved higher average final body weight (BW), comparable with that noted in the control group. Both RRC and FRC stimulated the antioxidant system of turkeys, which was reflected in a decrease in the concentrations of lipid hydroperoxides (LOOH) and malondialdehyde (MDA), and an increase in the total antioxidant potential (FRAP) and the concentration of total glutathione (GSH + GSSG) in blood plasma, compared with the control group. Turkeys fed diets with RRC were characterized by the highest blood vitamin C concentrations, the highest activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx), and lower catalase (CAT) activity. The dietary inclusion of rapeseed cake contributed to an increase in villus height and mucosal thickness in the duodenum, and a more beneficial influence was exerted by RRC. It can be concluded that the fermentation of rapeseed cake considerably reduces the concentrations of glucosinolates and phytate-phosphorus, and increases the final BW of turkeys. Diets containing 15% of RRC and FRC do not compromise metabolic parameters or immune function, and exert positive effects on antioxidant status and intestinal histomorphology in turkeys.

Propagation of an Avirulent Turkey Hemorrhagic Enteritis Virus Isolate in Chickens.

A series of studies were undertaken to optimize the propagation of hemorrhagic enteritis virus (HEV) in specific-pathogen-free (SPF) chickens. A total of 562 SPF chickens were orally inoculated with an Australian avirulent HEV isolate of turkey origin at 9, 14, 21, or 28 days of age with 5, 6, 7, or 8 log 10 genomic copies (GC), while 102 chickens served as uninfected controls. No clinical signs were observed in infected chickens. There was an inoculum-dose-dependent increase in the relative spleen and liver weight ( P < 0.01). Relative spleen weight 7 days post-HEV inoculation was up to 85% higher in chickens that were inoculated with 6 to 7 GC compared with controls, with no further increase at higher doses. Relative liver weight increased up to 14% in chickens inoculated with 6 GC, with no further increase. Birds inoculated with a 7 GC dose had a higher frequency of HEV DNA-positive birds (77% to 86%) than birds inoculated with lower doses (33% to 59%; P < 0.01). The most efficient dose for live passage propagation was 7 GC inoculated in 9-to-14-day-old birds, yielding an infection rate of 81%. Livers and spleens from infected birds at all doses were processed to produce a putative vaccine with a final GC recovery in the vaccine material of 8.6 GC/bird. In summary, HEV of turkey origin can be readily propagated in SPF chickens, and conditions to maximize viral retrieval were established.

La Sota vaccination may not protect against virus shedding and the lesions of velogenic Newcastle disease in commercial turkeys.

The aim of this project is to study the clinical signs and lesion of velogenic Newcastle disease (vND) in commercial turkeys, and also to find out if La Sota vaccination offered protection against these signs and lesions. The cockerels were included as positive controls. One hundred and twenty turkey poults and cockerels were divided into eight groups as follows: unvaccinated unchallenged turkeys (UUT), unvaccinated challenged turkeys (UCT), vaccinated unchallenged turkeys (VUT), vaccinated challenged turkeys (VCT), and along the same lines, the cockerel groups were UUC, UCC, VUC and vaccinated challenged cockerels (VCC). Vaccination was at 3 weeks of age while challenge was at 6 weeks of age. The unvaccinated turkeys and cockerels (UCT and UCC) showed different degrees of depression, diarrhoea and later paralysis at challenge. Total mortality was 100% in cockerels within 6 days, but 60% in turkeys. Similar but milder clinical signs were found in the VCC with a total mortality of 13.3%. The VCT showed mild drop in feed and water consumption, and no mortality. All the challenged groups had significant (p < 0.05) loss of weight when compared with their controls. Necropsy showed that while the UCC had severe proventricular haemorrhages, intestinal and caecal tonsil ulcers, the UCT had no digestive tract lesion. There was severe atrophy of the lymphoid organs in all the challenged groups. Histopathological sections of the bursa, spleen and thymus in all the challenged groups with special emphasis on the vaccinated and unvaccinated turkeys with mortalities of 0 and 60%, respectively, had very severe necrosis and depletion of the lymphoid tissue. Virus was isolated from the cloacal swabs. The haemagglutination inhibition antibodies were significantly higher (p < 0.05) in the challenged groups than the unchallenged. The above observations in the intestinal tracts of UCT are of diagnostic significance while the gross and microscopic lesions in the UCT and VCT show that La Sota vaccination may not protect turkeys against the destruction of the lymphoid organs by vND as earlier reported in chickens. This may lead to immunosuppression and production problems in areas where vND is enzootic.