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1.
Vet Res Commun ; 45(2-3): 101-109, 2021 Sep.
Article in English | MEDLINE | ID: mdl-33890234

ABSTRACT

The study investigated the mitigating effects of two probiotics on blood parameters of ISA Brown chicks inoculated with a very virulent infectious bursal disease virus (vvIBDV). Two hundred chicks were assigned into four groups of 50 birds each. Groups A and B were administered Antox® in water and Bactofort® in feed daily from 1 to 42 days of age and inoculated with a vvIBDV at 28 days and C and D served as positive and negative controls, respectively. Blood samples were examined for changes in packed cell volume (PCV), haemoglobin concentration (Hb), red blood cell (RBC), total white blood cell (TWBC), heterophil and lymphocyte counts seven days post inoculation. The PCV between groups A and C differed (P < 0.05) and in group B it was higher (P < 0.05) than that of group C. The Hb concentration between groups A, B and C differed (P < 0.05). There was a difference (P < 0.05) in RBC counts between groups A, B, C. Differences in TWBC between group A and C were significant (P < 0.05) and TWBC in group B was higher (P < 0.05) than that of group C. There was a significant difference in heterophil (P < 0.05) and lymphocyte (P < 0.05) count between group A and C, and B and C. Heterophil/lymphocyte ratio was significantly higher in positive control compared to groups A, B, C. Antox® and Bactofort® mitigated the deleterious effects of vvIBDV on blood parameters and can assist in cases of IBD outbreak.


Subject(s)
Birnaviridae Infections/veterinary , Chickens , Infectious bursal disease virus/physiology , Poultry Diseases/blood , Probiotics/administration & dosage , Animals , Birnaviridae Infections/blood , Birnaviridae Infections/physiopathology , Birnaviridae Infections/virology , Poultry Diseases/physiopathology , Poultry Diseases/virology
2.
Appl Microbiol Biotechnol ; 104(8): 3391-3402, 2020 Apr.
Article in English | MEDLINE | ID: mdl-32088761

ABSTRACT

Birnaviridae is a family of viruses (birnaviruses) which consists of four genera, members of which cause diseases in fish, birds, mollusks, and insects. The genome of birnaviruses encodes the highly immunogenic VP2 capsid protein. In order to demonstrate that the VP2 protein can be exploited as a diagnostic antigen for birnaviruses, we developed a lateral flow assay based on the surface-exposed VP2 protrusion domain of a representative birnavirus, infectious bursal disease virus (IBDV) of serotype 1 which causes the highly devastating infectious bursal disease in chickens. The biophysical characterization of the purified domain reveals that the domain predominantly consists of ß-sheets, exists in a trimeric form, and remains folded at high temperatures, making it suitable for diagnostic purposes. Owing to its highly immunogenic nature and excellent biophysical properties, we employed the VP2 protrusion domain in a gold nanoparticle-based lateral flow assay for rapid detection of anti-IBDV antibodies in serum samples of infected chickens. Our results indicate that the domain binds anti-IBDV antibodies with high specificity during laboratory testing and on-site testing. The lateral flow assay reported here yields comparable results in a qualitative manner as obtained through a commercial enzyme-linked immunosorbent assay (ELISA). As VP2 is a common capsid protein of birnaviruses, the lateral flow assay can be generalized for other birnaviruses, and members of Tetraviridae and Nodaviridae families which contain homologous VP2 capsid proteins.


Subject(s)
Antigens, Viral/immunology , Birnaviridae Infections/diagnosis , Birnaviridae Infections/veterinary , Birnaviridae/immunology , Viral Structural Proteins/immunology , Animals , Antibodies, Viral/blood , Antigens, Viral/genetics , Binding Sites, Antibody , Birnaviridae/genetics , Birnaviridae Infections/blood , Chickens/immunology , Circular Dichroism , Gold , Immunoassay , Metal Nanoparticles/chemistry , Poultry Diseases/diagnosis , Poultry Diseases/virology , Protein Domains , Spectroscopy, Fourier Transform Infrared , Viral Structural Proteins/genetics
3.
Vet Microbiol ; 229: 20-27, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30642595

ABSTRACT

The aim of the present study is to use Physalis mottle virus (PhMV) coat protein (CP) as a scaffold to display the neutralizing epitopes of Infectious bursal disease virus (IBDV) VP2. For this, three different chimeric constructs were synthesized by replacing the N-terminus of PhMV CP with tandem repeats of neutralizing epitopes of IBDV VP2 and expressed in Escherichia coli. Expression analysis revealed that all the three recombinant chimeric coat protein subunits are soluble in nature and self-assembled into virus-like particles (VLPs) as evidenced through sucrose density gradient ultracentrifugation. The chimeric VLPs were characterized by various biochemical and biophysical techniques and found that they are stable and structurally sound. When the chimeric VLPs were used as coating antigen, they were able to detect IBDV antibodies. These results indicated that the chimeric VLPs can be used as potential vaccine candidates for the control of IBDV, which needs to be further evaluated in animal models.


Subject(s)
Antibodies, Viral/blood , Birnaviridae Infections/veterinary , Infectious bursal disease virus/immunology , Animals , Antibodies, Neutralizing , Birnaviridae Infections/blood , Birnaviridae Infections/virology , Capsid Proteins/immunology , Capsid Proteins/metabolism , Chickens , Epitopes , Tymovirus , Viral Structural Proteins
4.
Poult Sci ; 98(1): 119-127, 2019 Jan 01.
Article in English | MEDLINE | ID: mdl-30107600

ABSTRACT

Ghrelin is a peptide hormone that plays a modulatory role in the immune system. Studies have demonstrated that mammal ghrelin level is influenced by pathological status. However, it has not been reported whether chicken ghrelin level changes during pathogen infection. This study was designed to investigate changes of ghrelin levels in chickens infected with infectious bursal disease virus (IBDV) and to explore the relationship between ghrelin changes and bursal damage, and inflammatory cells infiltration induced by IBDV. The results showed that (1) plasma ghrelin concentration increased after IBDV infection. It reached a peak at 10443.6 ± 2612.9 pg/mL on 2 dpi, which was about 100-fold as high as that of the control. Then it decreased sharply on 3 dpi, which was only 31.7% as that of 2 dpi, and remained stable until 5 dpi. Meanwhile, ghrelin and ghrelin-related gene, ghrelin-o-acyltransferase (GOAT), and growth hormone secretagogue receptor (GHSR) mRNA expression levels in bursa were also increased after IBDV infection, and reached the peak on 2 dpi at 149, 28.8, and 117.2-fold higher than that of the control, respectively. Then they decreased and remained at a higher status. Correlation analysis showed that plasma ghrelin concentration and ghrelin, GOAT, and GHSR mRNA expressions in bursa were strongly associated with IBDV VP2 mRNA expression in bursa. (2) The damage of bursa was the most severe on 5 dpi with a histopathological score of 12. It had no direct correlation with plasma ghrelin level and ghrelin, GOAT, and GHSR mRNA expressions in bursa. However, the number of inflammatory cells infiltrating into bursa, which was the highest on 2 and 3 dpi, showed significant a positive correlation with the ghrelin and GHSR mRNA expression. Presumably chicken ghrelin may function as an anti-inflammatory factor. In conclusion, IBDV infection upregulates the expression of ghrelin and ghrelin-related gene in chickens, and chicken ghrelin may play an important regulatory role during pathogen infection.


Subject(s)
Birnaviridae Infections/veterinary , Ghrelin/blood , Infectious bursal disease virus , Poultry Diseases/virology , Acyltransferases/genetics , Acyltransferases/metabolism , Animals , Birnaviridae Infections/blood , Birnaviridae Infections/virology , Bursa of Fabricius/enzymology , Bursa of Fabricius/pathology , Bursa of Fabricius/virology , Chickens , Gene Expression Profiling , Ghrelin/genetics , Poultry Diseases/blood , Poultry Diseases/metabolism , RNA, Messenger , Receptors, Ghrelin/genetics , Receptors, Ghrelin/metabolism
5.
Aust Vet J ; 96(5): 167-175, 2018 May.
Article in English | MEDLINE | ID: mdl-29691850

ABSTRACT

OBJECTIVE: To compare the effects of infectious bursal disease virus (IBDV) infection of commercial meat chickens at 0 and 16 days old (d.o.) and determine if IBDV vRNA is quantifiable in litter and dust samples. METHODS: Ross meat chickens (n = 60) were orally infected or not with IBDV at 0 or 16 d.o. Blood and faecal samples were collected longitudinally to 28 days post infection (dpi) from six chickens and tissues collected weekly from three euthanased chickens. Relative bursal weight was recorded postmortem. IBDV antibody titres in sera were measured using ELISA and VCN was determined in tissues, faeces, litter and dust using qRT-PCR. RESULTS: Chickens infected at 16 d.o. had earlier and more severe bursal atrophy, earlier and higher IBDV vRNA load in lymphoid organs and an earlier and greater antibody response to infection than those infected at 0 d.o. Faecal shedding of IBDV between 2 and 6 dpi was observed in both groups followed by cessation with the 0 d.o. group and re-initiation of shedding at 28 dpi. IBDV was readily detected and quantified in litter and dust samples. CONCLUSIONS: The presence of significant maternal antibody (MAb) titres in 0 d.o. chickens provided protection against IBDV replication and bursal atrophy at 7 and 14 days post infection. The reduced titres of MAb present at 16 d.o. did not prevent rapid IBDV replication and early marked bursal atrophy. The observed resistance of 0 d.o. chickens is likely to be a combination of MAb inhibition of IBDV and true age resistance of neonatal chicks. Measurement of IBDV in litter and dust may have research or diagnostic application.


Subject(s)
Birnaviridae Infections/veterinary , Chickens/virology , Infectious bursal disease virus/isolation & purification , Poultry Diseases/virology , Animals , Antibodies, Viral , Autopsy/veterinary , Birnaviridae Infections/blood , Birnaviridae Infections/immunology , Feces/virology , Food Microbiology , Genome, Viral , Infectious bursal disease virus/genetics , Likelihood Functions , Meat/virology , New South Wales , Poultry Diseases/blood , Tissue Distribution , Viral Load , Virus Shedding
6.
J Immunoassay Immunochem ; 36(4): 398-404, 2015.
Article in English | MEDLINE | ID: mdl-25331185

ABSTRACT

Infectious Bursal Disease Virus (IBDV) poses a great global threat to the poultry industry. Knowledge of the occurrence of the disease is important in the design and implementation of a control program, therefore this study determines the seroprevalence of IBDV in local chickens in Udu Local Government Area of Delta State. 250 chickens were bled by exsanguination and sera obtained were screened using Agar Gel Immunodiffusion (AGID) test. The seropositivity was 51.6%, which is indicates endemicity of the disease. Biosecurity and good sanitary measures are recommended. Molecular characterization of the strains should be carried out for inclusion in generic vaccines.


Subject(s)
Antibodies, Viral/blood , Birnaviridae Infections , Infectious bursal disease virus/immunology , Animals , Birnaviridae Infections/blood , Birnaviridae Infections/epidemiology , Birnaviridae Infections/immunology , Birnaviridae Infections/veterinary , Chickens , Nigeria/epidemiology , Seroepidemiologic Studies
7.
Poult Sci ; 92(4): 1040-9, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23472027

ABSTRACT

Based on the findings of a recent study suggesting a decreased cold-induced ascites incidence in broiler progeny from hyperthyroid (HYPER) breeder hens, and a controversy on the effects of hyperthyroidism on immunocompetence, the present study was conducted to determine the probable adverse effect of induced maternal hyperthyroidism on immune function in progeny chicks. Breeder hens (n = 88) were randomly allotted to the control or HYPER groups and received common or thyroxine (T4)-added (1 mg/L) water, respectively. The hens were artificially inseminated, and hatching eggs (n = 924) were incubated. Thereafter, the male hatchlings (n = 288) were reared for 42 d, and several cellular and humoral immune responses were evaluated at standard or low ambient temperature. Prevaccination antibody titers to Newcastle disease, infectious bronchitis, and infectious bursal disease virus were higher in HYPER chicks during 1 wk of age, although not different in their dams. For primary response to SRBC administered at 7 d of age, HYPER chicks recorded higher total, IgM (d 14), and IgG (d 21) anti-SRBC antibody titers. Higher cutaneous basophilic hypersensitivity response in HYPER chicks (d 10) was not observed at 35 d of age. Carbon clearance assay showed no difference, but in vitro lymphoproliferative response to concanavalin A was higher in 19-d-old HYPER chicks, independent of temperature treatment. An increase in lymphocyte percentage coincided with a decreased heterophil percentage and heterophil to lymphocyte ratio (d 14) in the HYPER group. The weight of lymphoid organs in progeny was not influenced by the oral exposure of dams to extra T4. Independent of T4 treatment, cold exposure was generally associated with decreased immune functions at early stages. The data suggested that oral exposure of broiler breeder hens to 1 mg/L of T4 not only had no adverse effect on immune function, but also modulated early adaptive immune responses in progeny chicks for which the causal mechanisms remain to be unraveled.


Subject(s)
Adaptive Immunity , Antibodies, Viral/blood , Chickens , Hyperthyroidism/veterinary , Immunity, Maternally-Acquired , Poultry Diseases/immunology , Thyroxine/administration & dosage , Animals , Antibodies, Viral/immunology , Birnaviridae Infections/blood , Birnaviridae Infections/immunology , Birnaviridae Infections/veterinary , Coronavirus Infections/blood , Coronavirus Infections/immunology , Coronavirus Infections/veterinary , Female , Hyperthyroidism/chemically induced , Male , Newcastle Disease/blood , Newcastle Disease/immunology , Poultry Diseases/blood
8.
Lipids Health Dis ; 11: 15, 2012 Jan 25.
Article in English | MEDLINE | ID: mdl-22273277

ABSTRACT

BACKGROUND: Infectious bursal disease (IBD) results in economic loss due to mortality, reduction in production efficiency and increasing the usage of antibiotics. This study was carried out to investigate the modulatory roles of dietary n-3 polyunsaturated fatty acids (PUFA) enrichment in immune response and performance of IBD challenged broiler chickens. METHODS: A total of 300 day old male broiler chicks were assigned to four dietary n-3 PUFA ascending levels as the treatment groups (T1: 0.5; T2: 8.0; T3: 11.5; T4: 16.5) using combinations of tuna oil and sunflower oil. All diets were isocaloric and isonitrogenous. On day 28, all birds were challenged with IBD virus. Antibody titer, cytokine production, bursa lesion pre and post-challenge and lymphoid organ weight were recorded. RESULTS: On d 42 the highest body weight was observed in the T2 and T3 and the lowest in T4 chickens. Feed conversion ratio of the T2 broilers was significantly better than the other groups. Although productive parameters were not responded to the dietary n-3 PUFA in a dose-dependent manner, spleen weight, IBD and Newcastle disease antibody titers and IL-2 and IFN-γ concentrations were constantly elevated by n-3 PUFA enrichment. CONCLUSIONS: Dietary n-3 PUFA enrichment may improve the immune response and IBD resistance, but the optimum performance does not coincide with the optimum immune response. It seems that dietary n-3 PUFA modulates the broiler chicken performance and immune response in a dose-dependent manner. Thus, a moderate level of dietary n-3 PUFA enrichment may help to put together the efficiency of performance and relative immune response enhancement in broiler chickens.


Subject(s)
Birnaviridae Infections/drug therapy , Chickens/immunology , Fatty Acids, Omega-3/therapeutic use , Fish Oils/therapeutic use , Immunologic Factors/therapeutic use , Infectious bursal disease virus , Plant Oils/therapeutic use , Poultry Diseases/drug therapy , Animal Feed , Animals , Avian Proteins , Birnaviridae Infections/blood , Birnaviridae Infections/immunology , Birnaviridae Infections/virology , Bursa of Fabricius/drug effects , Bursa of Fabricius/pathology , Bursa of Fabricius/virology , Cytokines/blood , Dietary Supplements , Fatty Acids, Omega-3/chemistry , Fatty Acids, Omega-3/pharmacology , Fish Oils/chemistry , Fish Oils/pharmacology , Immunologic Factors/chemistry , Immunologic Factors/pharmacology , Male , Organ Size/drug effects , Plant Oils/chemistry , Plant Oils/pharmacology , Poultry Diseases/blood , Poultry Diseases/immunology , Poultry Diseases/virology , Sunflower Oil , Viral Load
9.
J Sci Food Agric ; 91(10): 1737-45, 2011 Aug 15.
Article in English | MEDLINE | ID: mdl-21480270

ABSTRACT

BACKGROUND: A large number of plants and their isolates have been shown to potentiate immunity. Some plants exert anti-inflammatory and anti-stress effects, others hepatoprotective activity. In this study, 320 1-day-old broiler chicks were randomly divided into four major groups A, B, C and D and fed rations supplemented with 0, 15, 20 and 22.5 g Berberis lycium kg⁻¹ ration respectively. Each group was further divided into two subgroups, one vaccinated against Newcastle disease (ND) and infectious bursal disease (IBD), the other non-vaccinated. Antibody titre against IBD and ND, relative weight of lymphoid organs, post-challenge morbidity and mortality, serum hepatic enzymes and total serum protein were observed. RESULTS: Group C had higher anti-IBD and anti-ND antibody titres. Relative bursa weight in groups C and D was higher until day 28, but birds in group C performed better at later stages of examination. Relative spleen weight was highest in group C. During initial stages there was no effect on relative thymus weight, but at later stages the effect was significant. Groups C and D performed similarly in terms of relative thymus weight. The birds were challenged to field IBD through intramuscular injection at a dose rate of 0.5 mL per bird. Post-challenge morbidity was lowest in groups C and D, while treatment significantly (P < 0.001) affected mortality amongst affected (morbid) birds. Levels of serum alanine aminotransferase and alkaline phosphatase were lowest in group C. Serum protein was similar in all groups and in both vaccinated and non-vaccinated broiler chicks. CONCLUSION: Berberis lycium added to feed at 20 g kg⁻¹ is effective in improving immunity against ND and IBD as well as liver function in broiler chicks.


Subject(s)
Antibodies, Viral/blood , Berberis , Chickens/immunology , Immunologic Factors/pharmacology , Liver/drug effects , Plant Extracts/pharmacology , Poultry Diseases/immunology , Alanine Transaminase/blood , Alkaline Phosphatase/blood , Animals , Birnaviridae Infections/blood , Birnaviridae Infections/veterinary , Birnaviridae Infections/virology , Bursa of Fabricius/drug effects , Dietary Supplements , Infectious bursal disease virus/immunology , Liver/enzymology , Newcastle Disease/blood , Newcastle Disease/immunology , Newcastle Disease/virology , Newcastle disease virus/immunology , Organ Size/drug effects , Phytotherapy , Poultry Diseases/blood , Poultry Diseases/virology , Random Allocation , Spleen/drug effects , Thymus Gland/drug effects , Viral Vaccines
10.
J Vet Diagn Invest ; 23(2): 320-4, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21398455

ABSTRACT

The immunochromatographic assay is an alternative method for simple and rapid detection of Infectious bursal disease virus (IBDV) in chickens using colloidal gold-antibody conjugate. The whole-virus antigen of IBDV (UPM04190 isolate) and the high-affinity polyclonal antibodies directed against IBDV were blotted onto nitrocellulose membranes for test and control lines, respectively. Evaluation of the strip was performed using serum samples from experimentally and naturally infected chickens. The results showed that the test strip was more sensitive than the commercial enzyme-linked immunosorbent assay (ELISA) because it could detect a dilution factor up to 120,000 (250 ELISA units) for positive samples. It was also specific, in that it detected IBDV antibodies and did not cross-react with antibodies to other chicken viruses. The method was rapid (2 min) in both clinical and field environments with samples needing only a minimum amount (50 µl) of blood to produce an acceptable detection signal. The pen-site test strip proved successful in monitoring the immune status of chickens against the IBDV infection.


Subject(s)
Antibodies, Viral/blood , Birnaviridae Infections/veterinary , Chickens , Infectious bursal disease virus/isolation & purification , Poultry Diseases/virology , Reagent Strips , Animals , Antigens, Viral/chemistry , Birnaviridae Infections/blood , Birnaviridae Infections/virology , Metal Nanoparticles/chemistry , Poultry Diseases/blood , Sensitivity and Specificity
11.
J Virol Methods ; 165(2): 277-82, 2010 May.
Article in English | MEDLINE | ID: mdl-20152862

ABSTRACT

The routine technique for detecting antibodies specific to infectious bursal disease virus is a serological evaluation by enzyme linked immunosorbent assay (ELISA) with preparations of whole virions as antigens. To avoid the use of complete virus in the standard technique, in-house VP2 and VP3 based ELISAs were developed. Accordingly, four types of indirect ELISAs viz., a commercial IDEXX-ELISA kit, VP2 and or VP3 antigen based ELISAs and a whole virus ELISA were compared with the virus neutralization test. It was concluded that the sensitivity and specificity at receiver-operating characteristics (ROC) optimized cut-off of four ELISAs viz., IDEXX-ELISA, VP2-ELISA and VP3-ELISA indicated similar performance whereas whole virus antigen based ELISA showed poor performance in comparison to other ELISAs. Similarly the positive and negative likelihood ratio of four ELISAs at an optimized cut-off indicated IDEXX-ELISA to be the best among all the four ELISAs while the performance of rVP3-ELISA and rVP2-ELISA is good as compared to the whole virus ELISA. Finally, the area under the ROC curve (AUC) of four ELISAs which represented a summary statistics of the overall diagnostic performance of the test also indicated that the IDEXX-ELISA, VP3-ELISA and VP2-ELISA had similar and relatively better performance when compared to whole virus antigen-ELISA.


Subject(s)
Antibodies, Viral/blood , Birnaviridae Infections/veterinary , Enzyme-Linked Immunosorbent Assay/veterinary , Infectious bursal disease virus/isolation & purification , Poultry Diseases/diagnosis , Animals , Antigens, Viral/immunology , Birnaviridae Infections/blood , Birnaviridae Infections/diagnosis , Chickens , Infectious bursal disease virus/immunology , Poultry Diseases/blood , Recombinant Proteins/immunology , Sensitivity and Specificity , Viral Structural Proteins/immunology
12.
Avian Pathol ; 36(5): 367-74, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17899460

ABSTRACT

This study investigates the pivotal role of chicken interferon-gamma (ChIFNgamma) in the pathogenesis and immunosuppression of infectious bursal disease virus (IBDV) infection and is divided into in vivo, ex vivo and in vitro experiments. Two-week-old specific pathogen free chickens were inoculated with the 849VB very virulent strain of IBDV. The levels of systemic ChIFNgamma and chicken interleukin-6 in the serum were followed for 2 weeks during in vivo experiments. Then, splenocytes and bursal cells from infected chickens were analysed for their immunocompetence after mitogenic activation in ex vivo experiments. Finally, in vitro experiments were conducted to assess the direct immunosuppressive effect of ChIFNgamma on splenocytes and peripheral blood lymphocytes from non-inoculated specific pathogen free chickens. Our results reveal that the acute phase of infectious bursal disease coincides, on one hand, with high levels of systemic ChIFNgamma and chicken interleukin-6 and, on the other hand, with a strong inhibition of proliferation and activation of mitogen-stimulated splenocytes from infected chickens, as measured by ChIFNgamma production. Two weeks after viral inoculation, T lymphocytes infiltrating the bursa of Fabricius had recovered their activation capability. Finally, an in vitro study showed that the proliferation of naïve splenocytes and peripheral blood lymphocytes was directly and specifically inhibited by ChIFNgamma. In conclusion, a ChIFNgamma dysregulation occurs in chickens infected with IBDV and the overproduction of ChIFNgamma by T lymphocytes plays a key role in the pathogenesis and immunosuppression induced by this virus.


Subject(s)
Birnaviridae Infections/veterinary , Chickens/immunology , Infectious bursal disease virus , Interferon-gamma/metabolism , Animals , Birnaviridae Infections/blood , Birnaviridae Infections/immunology , Bursa of Fabricius/cytology , Bursa of Fabricius/immunology , Cells, Cultured , Immunosuppression Therapy , Lymphocytes/immunology , Lymphocytes/virology , Mitogens , Poultry Diseases/blood , Poultry Diseases/immunology , Specific Pathogen-Free Organisms , Spleen/cytology
13.
Avian Dis ; 50(2): 303-5, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16863087

ABSTRACT

The effect of a systemic disease on the dynamics of iron, zinc, and copper in chickens fed ad libitum was examined by infecting 10-day-old specific pathogen-free chickens with infectious bursal disease virus (IBDV). Liver, bursa of Fabricius, pancreas, spleen, and serum were sampled in 10 controls and 10 challenged chickens at 3-day intervals postinfection (PI) for 15 days. The samples were analyzed using atomic absorption spectroscopy. Serum levels were similar to that reported in the literature. Concentrations of iron and zinc did not change significantly in the pancreas, but there was an increase in copper in infected pancreatic tissue on days 9 and 15 PI. Iron concentration in the spleen showed a significant increase on days 6, 9, and 15 PI, whereas zinc was only significantly increased on day 15 PI. There was no significant change in copper concentrations in the spleens of infected chickens vs. controls. This finding is in line with previously reported data. The results showed that the liver was not a major tissue where iron and zinc were sequestered, as previous data have shown in mammals. Instead, the bursa of Fabricius had significantly increased levels of both iron and zinc in infected tissue vs. control tissue from 9 days PI on. Furthermore, the bursa had increased levels of copper in the latter portion of the study. These findings suggest that the bursa of Fabricius rather than the liver is the major organ for metallic ion sequestering during IBDV infection.


Subject(s)
Birnaviridae Infections/veterinary , Chickens/metabolism , Chickens/virology , Infectious bursal disease virus/pathogenicity , Metals/blood , Metals/metabolism , Poultry Diseases/metabolism , Animals , Birnaviridae Infections/blood , Birnaviridae Infections/metabolism , Bursa of Fabricius/chemistry , Bursa of Fabricius/metabolism , Chickens/blood , Copper/blood , Copper/metabolism , Iron/blood , Iron/metabolism , Liver/chemistry , Liver/metabolism , Pancreas/chemistry , Pancreas/metabolism , Poultry Diseases/blood , Poultry Diseases/virology , Specific Pathogen-Free Organisms , Spleen/chemistry , Spleen/metabolism , Zinc/blood , Zinc/metabolism
14.
Acta Vet Hung ; 53(4): 501-6, 2005.
Article in English | MEDLINE | ID: mdl-16363151

ABSTRACT

In the present study, specific-pathogen-free, 2-week-old Leghorn chickens were experimentally infected with strain 73688 of infectious bursal disease virus (IBDV) in order to evaluate haematological and histological changes that might suggest a pathomechanism for haemorrhages in this disease. At 96 hours post infection (hpi) a significant increase in prothrombin time was detected in the absence of visible lesions in myeloid bone marrow tissue and of significant thrombocytopenia. The aforementioned findings suggest alteration of the secondary coagulation mechanisms and not a direct effect of virus on thrombocytes or its precursors.


Subject(s)
Birnaviridae Infections/veterinary , Infectious bursal disease virus/classification , Infectious bursal disease virus/physiology , Poultry Diseases/blood , Poultry Diseases/pathology , Animals , Birnaviridae Infections/blood , Birnaviridae Infections/pathology , Birnaviridae Infections/virology , Chickens , Female , Infectious bursal disease virus/pathogenicity , Male , Poultry Diseases/virology
15.
Dis Aquat Organ ; 63(1): 43-52, 2005 Jan 25.
Article in English | MEDLINE | ID: mdl-15759799

ABSTRACT

We studied the characteristics of rainbow trout serum (RTS) inhibitory activity against infectious pancreatic necrosis virus (IPNV). Serum inhibition was related to the serum source and host cell in which the virus had been propagated. IPNV was more efficiently inhibited by RTS in salmonid cell lines than in non-salmonid cell lines, with inhibition highest in rainbow trout gonad (RTG)-2 cells. The RTS sensitivity of the virus was modified by the cell line through which the virus passed, with multiple passages through Chinook salmon embryo (CHSE)-214 cells producing a virus that was less sensitive to RTS. The RTS inhibition level was dependent on cell density: at a cell density of < or = 2 x 10(5) cells ml(-1), inhibition was insignificant (tissue culture infective dose 50% = 10(-1.1) TCID50 ml(-1) reduction); however, above a density of 3 x 10(5) cells ml(-1), the inhibition level was very high (> or = 10(-6.3) TCID50 ml(-1) reduction). The salmonid sera tested showed high inhibition, except for brook trout serum (BTS), while non-salmonid sera did not inhibit IPNV, replication on RTG-2 cells. Pretreatment of cultured cells with RTS prior to exposure did not affect inhibition of IPNV and thus did not mask a viral receptor. The RTS inhibition level was dependent on the time of serum addition, with inhibition being maintained for at least 16 h postinfection. Pretreatment of IPNV revealed that the virus is directly inhibited by RTS, and more strongly so when RTS is present during viral replication.


Subject(s)
Birnaviridae Infections/veterinary , Fish Diseases/immunology , Fish Diseases/virology , Infectious pancreatic necrosis virus/immunology , Oncorhynchus mykiss , Serum/immunology , Animals , Birnaviridae Infections/blood , Birnaviridae Infections/immunology , Cell Count , Cell Line , Fish Diseases/blood , Infectious pancreatic necrosis virus/growth & development , Species Specificity , Time Factors
16.
Vet Res Commun ; 28(4): 339-46, 2004 May.
Article in English | MEDLINE | ID: mdl-15222738

ABSTRACT

An immunocomb-based dot-ELISA, employing specially designed apparatus, was used to measure the antibody status for the three major poultry diseases--Newcastle disease, infectious bursal disease and infectious bronchitis--in single test sera. Positive samples could be classified into strong, moderate and weak positives by comparison with the colour reaction given by known strong and weak positive serum controls. The simultaneous dot-immunobinding assay gave reproducible results and allowed considerable savings on the cost of reagents compared to liquid ELISA. The antigen-coated immunocomb can be stored under refrigeration and the test can be performed rapidly under field conditions by trained personnel.


Subject(s)
Birnaviridae Infections/veterinary , Chickens , Coronavirus Infections/veterinary , Enzyme-Linked Immunosorbent Assay/veterinary , Newcastle Disease/blood , Poultry Diseases/virology , Animals , Antigens, Viral , Birnaviridae Infections/blood , Birnaviridae Infections/virology , Coronavirus Infections/blood , Coronavirus Infections/virology , Enzyme-Linked Immunosorbent Assay/instrumentation , Enzyme-Linked Immunosorbent Assay/methods , Infectious bronchitis virus/isolation & purification , Infectious bursal disease virus/isolation & purification , Newcastle Disease/virology , Newcastle disease virus/isolation & purification , Poultry Diseases/blood
17.
Vet Immunol Immunopathol ; 89(1-2): 67-81, 2002 Oct 08.
Article in English | MEDLINE | ID: mdl-12208052

ABSTRACT

In this report, we show that dimethylsulfoxide (DMSO) enhances liposome-mediated transfection of nucleic acid in chicken macrophage cells and that this could be exploited for the transcutaneous delivery of naked DNA through the intact skin of chickens. We found that DMSO enhanced transfection efficiencies of lipofectamine and polyethyleneimine in HD-11 chicken macrophage cells. Based on this principle, we showed that transcutaneous delivery of a DNA plasmid-dimethylsulfoxide mixture (1:1) to untreated skin of chickens results in a wide distribution of the plasmid in the body. Distribution studies were done using plasmids encoding enhanced green fluorescent protein (EGFP) reporter gene and a bivalent DNA vaccine coding for infectious bursal disease virus (IBDV) and Newcastle disease virus (NDV) immunogenic protein genes. This bivalent vaccine induced mucosal and systemic immune responses, as evidenced by IgA and IgM production in the tears and serum of vaccinated chickens. Mucosal immune responses in the tears after topical vaccination were significantly higher (P < 0.05) than after i.m. delivery of the same DNA vaccine and were characterized by the absence of an IgG response. The biodistribution of plasmid indicated that topical delivery with DMSO resulted in a wide distribution and persistence of the plasmid until 15 weeks post-primary vaccination. Both delivery methods resulted in insert-specific message being made in several body tissues, but after topical delivery the virus-specific mRNA could be detected in the bone marrow of one out of three chickens until 15 weeks post-primary vaccination. Furthermore, transcutaneous delivery of this DNA vaccine using DMSO conferred protection from challenge with virulent IBDV (86% survival) and NDV (86% survival). This novel transcutaneous method of delivery of a DNA vaccine shows promise as being an easy and effective way to deliver nucleic acids through intact skin for vaccination or therapeutic purposes.


Subject(s)
Chickens/immunology , Dimethyl Sulfoxide/administration & dosage , Immunization/veterinary , Plasmids/administration & dosage , Vaccines, DNA/administration & dosage , Administration, Cutaneous , Animals , Birnaviridae Infections/blood , Birnaviridae Infections/immunology , Birnaviridae Infections/veterinary , Birnaviridae Infections/virology , Chickens/virology , Immunization/methods , Immunoglobulin A/biosynthesis , Immunoglobulin A/blood , Immunoglobulin M/biosynthesis , Immunoglobulin M/blood , Infectious bursal disease virus/genetics , Infectious bursal disease virus/immunology , Injections, Intramuscular/veterinary , Neutralization Tests/veterinary , Newcastle Disease/blood , Newcastle Disease/immunology , Newcastle Disease/virology , Newcastle disease virus/genetics , Newcastle disease virus/immunology , Plasmids/genetics , Plasmids/pharmacokinetics , RNA, Viral/chemistry , RNA, Viral/genetics , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Specific Pathogen-Free Organisms , Tears/immunology , Tears/virology , Vaccines, DNA/immunology , Vaccines, DNA/standards
19.
J Virol Methods ; 85(1-2): 55-64, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10716338

ABSTRACT

A quantitative reverse transcription polymerase chain reaction (RT-PCR) protocol for assessing infectious bursal disease virus (IBDV) RNA levels in blood was developed using the ABI PRISM 7700 Sequence Detection System coupled with TaqMan chemistry. To control for variations in sampling and processing between samples 28S rRNA was co-amplified in a multiplex reaction and used to quantify total RNA. Relative quantification and standardisation was achieved using a log10 dilution series of RNA extracted from IBDV stock. A linear relationship was observed between input RNA and cycle threshold values (C(T)) over 5 log10 dilutions for the IBDV-specific product and 6 log10 dilutions for the 28S rRNA-specific product. As a test of the assay it was used to determine whether differences in susceptibility to IBDV observed between inbred lines of chickens could be detected at the level of viral load in the blood. Viral RNA levels peaked 2 days post-infection when there was significantly less viral RNA in the blood of resistant line 6(1) chickens compared with the more susceptible Brown Leghorns (P = 0.01). These results demonstrate that the course of IBDV infection can be monitored by quantifying IBDV RNA extracted from blood of infected chickens using TaqMan technology.


Subject(s)
Birnaviridae Infections/virology , Infectious bursal disease virus/genetics , RNA, Viral/blood , Animals , Birnaviridae Infections/blood , Bursa of Fabricius/virology , Chickens , RNA, Viral/analysis , Reverse Transcriptase Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/standards
20.
Vet Immunol Immunopathol ; 70(3-4): 309-16, 1999 Sep 20.
Article in English | MEDLINE | ID: mdl-10507370

ABSTRACT

Mannan-binding lectin (MBL) is a serum collectin which is believed to be an opsonin of the innate immune defence against various microorganisms. MBL is a minor acute phase reactant in man. We investigated the concentration of serum MBL in chickens infected with infectious bronchitis virus (IBV) and infectious laryngotracheitis virus (ILTV). The concentration of serum MBL increased about twofold (from approximately 6 to 12 microg/ml) due to these viral infections. The concentration peaked 3-7 days after infection with IBV, and 3-5 days after ILTV infection, depending on the ILTV strain used. The increased levels returned to normal values 6-10 days after infection. The results indicated that MBL is a minor acute phase reactant in chickens.


Subject(s)
Birnaviridae Infections/blood , Carrier Proteins/blood , Herpesviridae Infections/blood , Lectins/blood , Mannans/blood , Poultry Diseases/blood , Animals , Chickens , Collectins , Enzyme-Linked Immunosorbent Assay/veterinary , Herpesvirus 1, Gallid , Infectious bursal disease virus
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