Comparison of Chicken Immune Responses to Immunization with Vaccine La Sota or ZG1999HDS Strain of Newcastle Disease Virus.

Newcastle disease (ND) is a highly contagious avian disease. Global control of ND is mainly based on vaccination of poultry; however, reported outbreaks of ND in vaccinated flocks indicate a constant need to re-evaluate the existing vaccines and a development of the new ones. In this study, 4-week-old male chickens of the layer commercial hybrid were immunized oculonasally with a commercial NDV live La Sota vaccine (LS group), a suspension of lyophilized NDV strain ZG1999HDS (ZG group), or saline (Control (K) group). Antibody response was determined by haemagglutination inhibition (HI) assay. Cell-mediated immunity (CMI) was characterized by immunophenotyping of leukocyte's and T-lymphocyte's subpopulations (flow cytometry). Applied NDV strains did not cause any adverse reaction in treated chickens. Both strains induced the significantly higher HI antibody response in comparison to the control group, and overall antibody titer was higher in ZG group than in LS group. CMI, manifested as a higher proliferation of B- and T-helper cells, yielded better results in the ZG groups than in the LS group. Based on the obtained results, we conclude that the strain ZG1999HDS is immunogenic and is a suitable candidate for further research and development of poultry vaccines.

The Enhancing Effects of 10% PBS Washout of Holocene Minerals on HuIFN-αN3 Inducing Capacity of NDV ZG1999HDS or Sendai virus (Cantell Strain).

Different strains of Newcastle disease viruses (NDV) or Sendai viruses (SV) are used to induce the production of human leukocyte multi subtype interferon-alpha (HuIFN-αN3). Their inducing capacity can be enhanced in different ways. One includes 10% PBS washout of Holocene minerals (HM). The presented study aims to compare the HuIFN-αN3 inducing capacity of NDV ZG1999HDS or SV (Cantell strain) strain in vitro, and to evaluate the enhancing effect of 10% PBS washouts of HM on both viruses. The NDV strains' ZG1999HDS interferon inducing capacity (483.23 ± 4.5 pg/mL) was similar to that of the SV (Cantell strain) (584.16 ± 5.9 pg/mL). It was shown that the HuIFN-αN3 inducing capacity of the strain of NDV ZG1999HDS can be strongly enhanced with 10% PBS washout of HM to 3818.21 ± 41.9 pg/mL and 4790.34 ± 33.5 pg/mL with SV (Cantell strain), u. The RP-HPLC analyses of such HuIFN-αN3 induced with the strain of NDV ZG1999HDS show the difference to SV (Cantell strain) induced HuIFN-αN3 in the absence of subtype α14 and the lower level of the subtype α1. The possible ways of such enhancement were also studied and it was postulated that the Fe2+ ions from 10% PBS washouts of HM, while stimulating the reactive oxygen species (ROS) and nitric oxide (NO) formation, activate the transcription factor NF- κB and consequently the production of HuIFN-αN3.

Immunogenicity of Newcastle disease virus strain ZG1999HDS applied oculonasally or by means of nebulization to day-old chicks.

Newcastle disease (ND) is one of the classic viral infections of poultry which resists all the efforts of eradication. Newcastle disease virus (NDV) strain ZG1999HDS was isolated during the outbreak in 1,999 at a broiler farm in Croatia. Previous trials in chickens confirmed it to be a lentogenic pathotype and immunogenic by stimulating humoral and cell mediated immunity. Further characterization by deduced amino acid sequence at the cleavage site of fusion protein confirmed its lentogenic nature, and in vitro tests its oncolytic capacity. Owing to its immunogenicity, strain ZG1999HDS is considered for vaccine development. In this study, 1-day-old chicks were vaccinated using strain ZG1999HDS oculonasally or by nebulization. Strain ZG1999HDS induced humoral immune response in both immunized groups The cell-mediated immune response occurred earlier in the group immunized by nebulization, as shown by a higher frequency rate of T and B lymphocytes, and significantly higher expression of IFN-α in respiratory organs and IFN-γ expression in the spleen. Viral genomic RNA was not detected in investigated organs. Thus, NDV strain ZG1999HDS is immunogenic when administered by means of nebulization or oculonasally without any adverse effects and is therefore suitable for further research and vaccine development. Further research is needed regarding its tropism.

Immunogenicity and safety of Queensland V4 and Ulster 2C strains of Newcastle disease virus given to maternally immune, newly hatched chickens by nebulization.

Commercial chickens with a high level of maternal antibodies for Newcastle disease were vaccinated when newly hatched with Queensland V4 or Ulster 2C Newcastle disease virus (NDV) strains by nebulization. The exposure time to a fine aerosol of vaccine produced with an ultrasonic nebulizer was 60 sec. The chickens were challenged oculonasally with virulent NDV strain Texas GB in weekly intervals up to the 49th day of life. Although protected for several weeks by maternal antibody, they were sufficiently protected thereafter by active immune response to the vaccines. Vaccinal reactions were not observed. Queensland V4 produced higher titers than Ulster 2C and provided better protection to challenge.

Enhancing Effect of 100.414-kHz Electromagnetic Field Produced by Defender's Pulse Generator on the ChIFN γ-Like Molecule Inducing Capacity of Lens culinaris Agglutinin and 10% PBS Washouts of Different Holocene Minerals.

Macrophages play key role in host defense and tissue repair, and thus understanding regulation of their function is important. For instance, our previous results have shown that in chicken macrophage system (CoMA cell line), application of a pulse of electromagnetic fields of frequencies 0.618, 1.054, 5.229, and 100.414 kHz induces production of interferon γ-like molecules. In this study, we have shown that the electromagnetic field of 100.414 kHz is the most effective in inducing synthesis of chicken interferon γ and chicken interferon γ-like molecules in CoMA cells, especially when combined with Lens culinaris agglutinin and 10% phosphate-buffered saline washouts of different Holocene minerals. A 2-minute pulse of electromagnetic field was produced by Defender's pulse generator. Both chicken interferon γ and chicken interferon γ-like molecules from the cell supernatant were evaluated by an antiviral assay and were also analyzed with reverse-phase high-performance liquid chromatography on Phenomenex, Aeris peptide columns. Our results show that application of a single inducing factor ( Lens culinaris agglutinin, 100.414 kHz electromagnetic field, 10% phosphate buffer saline washout) or combined usage of 2 of them moderately stimulated production of chicken interferon γ-like molecules (from 1.550 to 48.028 IU/mL), whereas the combination of 10% phosphate-buffered saline washout of Koprivnica rock + Lens culinaris agglutinin + 100.414 kHz/9 V resulted in an output of 162.122 IU/mL. Hence, we may conclude that a combined use of electromagnetic field, Holocene minerals, and Lens culinaris agglutinin greatly stimulates synthesis of chicken interferon γ-like molecules in CoMA cells.

Enrichment of eggs in n-3 polyunsaturated fatty acids by feeding hens with different amount of linseed oil in diet.

The production of n-3 polyunsaturated fatty acid enriched eggs by addition of linseed oil to the laying hens' diet has been evaluated in terms of production parameters and n-6/n-3 ratio. A total of 150 18weeks old Lohmann Brown laying hens were housed in cages and fed with basal diet and four experimental diets containing 1%, 2%, 3% or 4% of linseed oil added to the basal diet. The effect of the altered level of linseed oil on hens laying performance, fatty acid content and composition and cholesterol content in egg yolk has been evaluated during 13weeks of experiment. Egg weight, yolk fat content, yolk weight, yolk percentage and shape index were not influenced by dietary treatment. The ratio between n-6 and n-3 polyunsaturated fatty acids in eggs decreased in first 5weeks and then remained stable until the end of the experiment for all experimental groups. Different contents of linseed oil in feed highly influenced the n-6/n-3 ratio (P<0.0001). Addition of linseed oil did not influence the cholesterol content in yolks (P=0.5200) while the only factor affecting the cholesterol content was the hens age (P<0.0001).