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.
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.
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 Fe
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.
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.
► Addition of linseed oil in feed did not influence the egg production parameters. ► Linseed oil significantly affected the ratio of n-6/n-3 FAs in eggs. ► Shortly after adding linseed oil into feed, ...the reduction of the ratio was noticed. ► The cholesterol content wasn’t influenced by addition of linseed oil in feed.
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).
Marek's disease is a common lymphoproliferative disease of chickens, usually characterized by mononuclear cell infiltration of peripheral nerves and various other organs and tissues, including the ...iris and skin. A group of 70 newly-hatched chicks was vaccinated by means of nebulization and exposed for 60 seconds to the HVT FC 126 vaccine, while the other group of 70 chicks received the same vaccine by parenteral route (s.c.). The aim of this study was to compare the morphological patterns of the chicken's lymphocytes in their peripheral blood, before and after vaccination by means of either nebulization or parenteral injection. Image analysis was performed using the SFORM software (VAMSTEC, Zagreb, Croatia). A total of 50 blood smears from vaccinated chickens (20 by means of nebulization and 30 by parenteral injection), with an average of 100 cells per smear, were analyzed. The results showed that the peripheral blood lymphocytes of chickens vaccinated by means of nebulization, compared with parenteral vaccinated, had significantly higher values for the majority of the measured variables: area, outline, minimum and maximum radius, length, breadth and convex area, on all days after vaccination, except on day 4, indicating the significantly higher metabolic activity of those cells. In lymphocytes of chickens vaccinated parenterally, only the nucleo-cytoplasmic ratio was higher. The results show that morphological patterns of immunocompetent cell transformation could be used to evaluate immune responses to vaccination and vaccine efficacy. We conclude that nebulization as a mode of vaccination against Marek's disease stimulates the transformation of immunocompetent cells much earlier, thus shortening the time of immunosuppression and improving the immune response. This is of paramount importance for the practical application of Marek's disease vaccine. Key words: chickens, Marek's disease, nebulization, parenteral vaccination, lymphocyte morphometry Marekova je bolest limfoproliferativna zarazna bolest kokosi, obicno karakakterizirana mononuklearnim stanicnim infiltacijama u perifernim zivcima i ostalim organima i tkivima, ukljucujuci sarenicu i kozu. Skupina od 70 netom izleglih pilica cijepljena je protiv Marekove bolesti cjepivom od soja FC 126 herpesvirusa purana postupkom nebulizacije izlaganjem aerosolu tijekom 60 sekundi, dok je druga skupina od 70 pilica primila isto cjepivo parenteralnim nacinom (supkutano). Cilj ovog istrazivanja bio je usporediti morfometrijske znacajke limfocita u perifernoj krvi pilica prije i nakon cijepljenja kako postupkom nebulizacije tako i parenteralnim nacinom primjene. Kompjutorska analiza slike ucinjena je na osobnom racunalu koristenjem programa "SFORM" (VAMSTEC, Zagreb, Hrvatska). Ukupno je pretrazeno 50 obojenih razmaza periferne krvi cijepljenih pilica (20 postupkom nebulizacije i 30 parenteralno), te je analizirano prosjecno 100 limfocita po uzorku. Rezultati su pokazali da su limfociti u perifernoj krvi pilica cijepljenih postupkom nebulizacije u odnosu na parenteralno cijepljene bili znacajno veci u vecini istrazivanih pokazatelja: povrsini, opsegu, minimalnom i maksimalnom polumjeru, duzini i sirini, te ispupcenosti odnosno konveksnosti povrsine, u svim danima pokusa osim cetvrtoga, sto ukazuje na pojacanu metabolicku aktivnost tih stanica. U limfocitima parenteralno cijepljenih pilica samo je omjer jezgre i citoplazme bio veci. Postignuti rezultati pokazuju morfoloske znacajke transformacije imunokompetentnih stanica, koje se mogu rabiti u procjeni imunosnog odziva na cjepivo, ucinka cijepljenja i moguceg docjepljivanja. Zakljucili smo da nebulizacija kao metoda cijepljenja protiv Marekove bolesti stimulira transformaciju imunokompetentnih stanica mnogo ranije, skracujuci time vrijeme imunosupresije i pojacavajuci imunosni odziv. To je od osobite vaznosti za prakticnu primjenu cjepiva protiv Marekove bolesti. Kljucne rijeci : pilici, Marekova bolest, nebulizacija, parenteralno cijepljenje, morfometrija limfocita
Marek's disease is one of 'the greatest problems in poultry production 'today and vaccination is one of 'the most important ways of prevention. An innvoative method of vaccine delivery by means of ...nebulisation was used for vaccination of newly hatched chicks using HVT FC 126 and its impact on blood cell count (WBC, RBC and differential cell count) and H/L ratio was measured. The trial was performed from day 1 to day 21 of life of newly hatched male chicks. Standard blood cell count was performed in a Neubauer hemocytometer, and differential cell count on blood smears stained with MayGrunwald-Giemsa. Blood for that purpose was taken from the jugular vein. The results show a significantly higher WBC count in the group vaccinated by means of nebulisation than in the non-vaccinated groups on day 5 of the trial, and from the group that received only physiological solution, on day 14. In the RBC count there were fluctuations but without any significant differences between the groups during the trial. In differential blood cell count, there were some significant differences on days 7 and 21 of the trial, but no differences in H/L ratio. These results show that vaccination by means of nebulisation significantly influenced the blood cell count but all changes were within the physiological range. Nebulisation as a method of vaccination could probably improve the immune response to the wild Marek's disease virus, mimicking natural infection via the respiratory system and as a mass form of vaccination could be a powerful method for delivery of recombinant HVT vaccine. Key words: chicleen, Marek's disease, blood cell count, nebulisation, vaccination Marekova bolest predstavlja jedan od najvecih problema u uzgoju peradi pri cemu je cijepljenje jedan od najvaznijih postupaka suzbijanja. Koristen je novi postupak primjene cjepiva od soja FC 126 herpesvirusa purana jednodnevnim pilicima postupkom nebulizacije pri cemu je analiziran njegov utjecaj na broj stanica u krvi (broj leukocita, broj eritrocita i diferencijalnu krvnu sliku) kao i omjer neutrofila i limfocita (N/L). Pokus je proveden na jednodnevnim muskim pilicima od prvog do 21. dana zivota. Koristen je standardni postupak brojenja krvih stanica u Neubauerovoj komorici, dok je DKS nacinjen na razmazima pune krvi obojene MayGrunwald-Giemsom. Krv za navedene postupke uzeta je iz jugularne vene. Rezultati pokazuju znacajan porast broja leukocita u krvi kod skupine cijepljene postupkom nebulizacije u odnosu na necijepljene skupine petog dana pokusa, dok od skupine koja je primila samo fiziolosku otopinu ima znacajno vise leukocita 14. dana pokusa. Broj eritrocita pokazuje varijacije u pokusnih skupina, ali bez znacajnih razlika tijekom Citavog pokusa. Znacajne razlike u diferencijalnoj krvnoj slici bile su ustanovljene 7. i 21. dana pokusa, dok kod N/L omjera nije bilo znacajnih razlika. Navedeni rezultati pokazuju da je cijepljenje postupkom nebulizacije znacajno utjecalo na broj krvnih stanica te moguce potaknulo specifican imunosni odgovor, ali su sve promjene bile u fizioloskom rasponu. Nebulizacija kao postupak primjene cjepiva, iz navedenog, pretpostavlja se moze poboljsati imunosni odgovor na virus Marekove bolesti oponasajuci prirodan nacin zarzavanja putem disnog sustava, a moze biti izvrstan u masovnoj primjeni rekombinantnih cjepiva pripravljenih od herpesvirusa purana. Kljucne rijeci: pilici, Marekova bolest, broj krvnih stanica, nebulizacija, cijepljenje
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