PURPOSE: House dust mites (HDMs) are an important source of indoor allergens associated with asthma, rhinitis and atopic dermatitis. Chicken immunoglobulin (Ig) Y is known to be a good alternative to mice and rabbit antibody production. In this study, we produced IgYs specific to HDMs and investigated their IgE immunoreactivities. MATERIALS AND METHODS: Total IgYs were isolated from the yolks of White Leghorn hens immunized with either Dermatophagoides pteronyssinus or D. farinae protein extract. Control antibodies were separated from the yolks of immunized hens with phosphate buffered saline. IgYs specific to HDMs were analyzed using enzyme-linked immunosorbent assay and Western blotting analysis. RESULTS: The concentration of egg IgY specific to D. farinae in an immunized hen increased and the highest achieved was 661.3 ug/mg (per an egg) on day 47, compared with 760 ug/mg IgY specific to D. pteronyssinus on day 16. The D. pteronyssinus or D. farinae-specific IgY was detected by binding of each mite proteins, and their immunoreactivities were elevated dependent of the specific IgY concentration. CONCLUSION: IgY specific to HDMs may be a promising antibody for immunological diagnosis as well as identification of possible resistance relating to HDM allergy.
Allergens/*immunology ; Animals ; Antibodies/*immunology ; Chickens ; Egg Yolk/*immunology ; Female ; Immunoglobulins/*immunology ; Pyroglyphidae/*immunology

IgY antibodies, also called egg yolk immunoglobulins, are the only immunoglobulins in egg yolk and transferred in the female from serum to egg yolk to confer passive immunity to embryos and neonates. Using hens instead of mammals as the immunization host brings a number of advantages: Eggs are cheap and readily available; antibody levels in yolks are high; IgY isolation is fast and simple; and what is more; IgY neither binds the rheumatoid factor nor reacts to the mammalian complement factor. All these differences make IgY technology more widely applicable, such as in the production of polyclonal antibodies against various antigens, immunodiagnostics and immunotherapy, and many medical areas in both animals and human. IgY also has a good prospect in human immunocontraception.
Animals ; Chickens ; Egg Yolk ; immunology ; Female ; Humans ; Immunoglobulins ; isolation & purification ; therapeutic use ; Vitellogenins ; immunology

OBJECTIVETo prepare artificial antigens and anti-citrinin egg yolk-derived immunoglobulin (IgY) to build an enzyme-linked immunosorbent assay (ELISA) for citrinin (CTN).

METHODSCTN was conjugated with bovine serum albumin (BSA), ovalbumin (OVA) with formaldehyde condensation method to prepare artificial antigens and identified by ultraviolet (UV) spectrometry and Infrared (IR) spectrometry. Artificial antigens for CTN and anti-CTN IgY were purified with polyethylene glycol two-step precipitation method and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). ELISA with IgY was established. Cross-reactivity of IgY with various structural similarities to CTN and possible co-occurrence with CTN in agricultural commodities were studied.

RESULTSUV and IR absorption spectra suggested that CTN was correlated with the carrier protein of BSA or OVA. SDS-PAGE patterns showed that the anti-CTN IgY was almost pure with a molecular weight of approximate 100 KD. The indirect competitive ELISA showed that the detection limit of CTN was 10 ng x mL(-1), with a good linearity ranging 20-640 ng x mL(-1).

CONCLUSIONArtificial antigens of CTN can be successfully synthesized. The established ELISA can be used to determine CTN- contaminated samples.

Animals ; Antibody Specificity ; Antigens ; chemistry ; Chickens ; Citrinin ; chemistry ; Egg Yolk ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Immunoglobulins ; immunology

AIMTo improve specificity and accuracy of endogenous ouabain measurement assay.

METHODSAnti-ouabain polyclonal antibody egg yolk (IgY) and anti-ouabain rabbit antibody (IgG) were prepared respectively. In the presence of two kinds of antibody, then the specificity and accuracy of enzyme-linked immunosorbent assay (ELISA) were compared.

RESULTSThe ELISA, in the presence of IgY, provided a sensitivity of the average intraassay coefficient of variation(CV) was 2.03%, and the inter-assay CV was 2.34% respectively. In contrast, IgG were 2.83% and 3.29%. No significant interferences were observed with hydrocortisone and dexamethasone. There was 3.45% vs. 5.95%, 3.20% vs. 5.20% of crossreaction with cedilanid and digoxin.

CONCLUSIONThe specificity and accuracy of ELISA, in which IgY was used, were more better than IgG.

Animals ; Antibody Specificity ; Chickens ; immunology ; Cross Reactions ; Egg Yolk ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Immunoglobulin G ; immunology ; Immunoglobulins ; immunology ; Male ; Ouabain ; analysis ; Rabbits

OBJECTIVETo study the effects of different administrations of antibodies against adipocyte plasma membrane proteins on growth and fat deposition in rats.

METHODNinety six female SD rats that weighed approximately 140 g were allotted randomly into four groups which were given negitive control yolk and positive yolk containing antibody (IgY) against adipocyte plasma membrane (APM) proteins by intragastric administration (i.g.) and subcutaneous injection (s.c.) respectively. Rats were given 1 ml of yolk for every three days in i.g. groups. Rats were given 1 ml of yolk for 4 consecutive days, and the procedure was repeated after one month. The trial lasted for 75 days after which rats were slaughtered for carcass analysis and sampling.

RESULTSThe body weight gain and food intake of rats were not different between treatment and control groups. In i.g. goups, positive yolk decreased mesemteric fat index, paramertrial fat index and perirenal fat index (P < 0.05), reduced serum triglycerides (P < 0.05) and increased serum free fatty acids (FFA) (P < 0.01), and also decreased serum leptin, insulin and TNF-alpha levels ( P < 0.01 or P < 0.05), but did not affect gastrocnemius muscle index and serum cholesterol. In s.c. groups, positive yolk increased gastrocnemius muscle index (P < 0.05), reduced serum triglycerides and serum leptin (P < 0.01), increased serum TNF-alpha (P < 0.05), but did not affect adipose tissue depots, serum FFA, cholesterol and insulin.

CONCLUSIONAdministration of yolk antibody against APM proteins could effectively improve body composition of rats, and the treatment by intragastric administration could give better effect than by subcutaneous injection.

Adipose Tissue ; growth & development ; Animals ; Antibodies ; pharmacology ; Body Composition ; Egg Yolk ; immunology ; Female ; Growth and Development ; physiology ; Immunization ; Membrane Glycoproteins ; immunology ; Rats ; Rats, Sprague-Dawley

Swine respiratory diseases induce severe economic losses in the swine industry worldwide. Several methods have been developed and applied to control these diseases. However, there are still problems of disease control in the swine industry. Recently, egg yolk antibodies have been found to offer several advantages for disease control in animals and humans. In a previous study (24), antibodies to several causative pathogens of swine respiratory diseases were developed. However, several problems remained, especially in terms of reduced laying rates. Therefore, experimental vaccines were reformulated with various bacterial antigens of the swine respiratory diseases. After immunizing hens with the antigens, antibody profiles and other effects including laying rates were investigated and compared to those of the previous study. Profiles of antibody titers were very similar with those of the previous study. However, side effects, such as depression, weakness, reduction of laying rates and mortality, were dramatically lowered and laying rates were increased in hens injected with certain experimental vaccines. In particular, laying rates of hens injected with vaccines against atrophic rhinitis were increased up to 84% by injecting a vaccine composed of only the DNTs of B. bronchiseptica and P. multocida D:4. Efficacies of the vaccines against swine pneumonic pasteurellosis and pleuropneumonia were very similar with those of the previous study. These results suggest that new vaccines could be effective in the production of egg yolk antibodies against the causative agents of swine respiratory diseases.
Actinobacillus pleuropneumoniae/classification/genetics/*immunology ; Animals ; Antibodies, Bacterial ; Antibody Formation ; Bacterial Outer Membrane Proteins/genetics/isolation & purification ; Bordetella bronchiseptica/classification/genetics/*immunology ; Egg Yolk/microbiology ; Female ; Immunoglobulins/*genetics ; Oviposition ; Pasteurella multocida/classification/genetics/*immunology ; Serotyping ; Swine

Infectious bursal disease (IBD) is an acute and highly contagious disease of young chickens caused by Birnavirus. Mortality of infected birds can be best prevented if injected with antibodies. The present study was an attempt to raise specific hyper-immune polyclonal antibodies against IBD virus in Pakistan. Commercial layers divided into four groups were injected with IBD vaccine subcutaneously according to four different treatment regimens. Eggs were collected daily and antibodies were purified from yolk with dextran sulphate. Titers of antibodies in serum and yolk were evaluated with enzyme linked immunosorbant assay and agar gel precipitation test. Antibody titers were significantly higher in yolk than serum. Eggs collected at 28 days post-vaccination had maximum antibody titers. Of treatment regimens, T3 was found to be most effective for hyperimmunization. Lyophilized antibodies stored at 4oC did not lose their activity till the end of experiment. IBD virus infected birds were injected with purified antibodies which induced 92% recovery as compared to control birds. The study implicates that the purified antibodies may be useful as a therapeutic agent to cure IBD infected birds.
Animals ; Antibodies, Viral/blood ; Birnaviridae Infections/immunology/*therapy/*veterinary/virology ; *Chickens ; Egg Yolk/immunology/virology ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Immunization/methods/*veterinary ; Immunoglobulins/*immunology ; Immunotherapy/methods/veterinary ; Infectious bursal disease virus/*immunology ; Poultry Diseases/immunology/*therapy/*virology ; Precipitin Tests/veterinary ; Viral Vaccines/*immunology/therapeutic use

Specific immunoglobulin (IgY) from egg yolk against Aeromonas hydrophila was produced by immunization of White Leghorn hens with formalin-killed whole cells of A. hydrophila. ELISA test using A. hydrophila as the coating antigen revealed that the specific antibody titer started to increase in the egg yolk at the 13th day post-immunization (P/N=2.18), reached the peak at the 56th day (P/N=13.82), and remained at high level until day 133 (P/N=7.03). The antibody was purified by saturated ammonium sulphate with a recovery rate of 63.5%. The specific IgY inhibited the growth of A. hydrophila at a concentration of 1.0 mg/ml during the 18 h incubation. Pre-treatment of polyploid gibel carps Carassius auratus Gibelio with specific IgY had a protection rate of 60% (6/10) against challenge with A. hydrophila, while none of the fishes in the control groups receiving sterile phosphate buffered saline (PBS) or non-specific IgY survived the challenge. Treatment of fishes with the specific IgY 4 h after the challenge also had lower mortality (70%, 7/10), a 30% reduction against the control PBS or non-specific IgY groups (10/10). These results indicate that specific IgY antibodies could be obtained easily from hens immunized with an inactivated A. hydrophila and could provide a novel alternative approach to control of diseases in fishes caused by this organism.
Aeromonas hydrophila ; drug effects ; growth & development ; immunology ; Animals ; Antibody Specificity ; Antigen-Antibody Reactions ; Chickens ; immunology ; Dose-Response Relationship, Drug ; Egg Yolk ; chemistry ; Goldfish ; immunology ; microbiology ; Gram-Negative Bacterial Infections ; immunology ; prevention & control ; Immunoglobulins ; pharmacology ; therapeutic use ; Microbial Sensitivity Tests ; Survival Rate ; Time Factors