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

OBJECTIVETo obtain egg yolk antibody in hen eggs laid by hens immunized with the protein of Porphyromonas gingivalis (Pg). To generate, purify IgY against Pg (anti-Pg-IgY) and identify its specificity.

METHODSPgATCC33277 was cultured under standard anaerobic conditions and harvested after proliferation. Then Pg was extracted by sonication until the cell pellets were shattered completely. After centrifugatiton, the supernatant was collected. Five-month-old Roman hens were immunized for egg antibody production. The antibody was inoculated intramuscularly and subcutaneously in the breast from multiple spot with 1.0 ml of a vaccine consisting of oil-adjuvant protein which was mixed with 1 ml protein of Pg and 1 ml Freund's adjuvant complete every 10 days, for 4 times. The eggs were collected after the first immunization and stored at 4°C. The anti-Pg-IgY was extracted and purified. The protein concentration was tested by bicinchoninic acid (BCA), the specificity of IgY analyzed by SDS polyacrylamide gel electrophoresis (SDS-PAGE), the titre of IgY and its physicochemical character were evaluated by indirect enzyme-linked immunosorbent assay.

RESULTSThe concentration of obtained anti-Pg-IgY was 2.05 g/L. SDS-PAGE analysis of the anti-Pg-IgY showed that the molecular weight of IgY was consistent with the theoretical value. Protein of anti-Pg-IgY appeared approximately 5 days after the first immunization, and reached the peak at 50 - 55 days. Antibody titres reached 1:100 000. Each egg produced more than 10 mg IgY, and its purification was up to 95% as well.

CONCLUSIONSLayer hens immuned by Pg may provide specific IgY of high titre and high concentration. The antibody has high purity and is heat, acid and alkali-resistant.

Animals ; Antibodies ; chemistry ; immunology ; Chickens ; immunology ; Egg Yolk ; chemistry ; immunology ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Immunization ; Immunoglobulins ; chemistry ; immunology ; isolation & purification ; Porphyromonas gingivalis ; immunology

OBJECTIVETo prepare the oral solution of egg yolk hepatitis B virus (HBV)-specific transfer factor (EYHBV-TF) and evaluate its immunological activity as an immune regulator against hepatitis B.

METHODSFrom hens immunized with the Hepatitis B vaccine the egg yolk was isolated to extract the specific transfer factor EYHBV-TF, and its physicochemical properties were examined. Leukocyte adhesion inhibition test (LAI) was performed to detect the immunogenic activity of EYHBV-TF. The solution of EYHBV-TF was then administered orally in normal mice, and the specific cellular immune activity induced was assayed with delayed type skin hypersensitivity test (DTH), with the non-specific immune activity assessed with immune organ index. The immune responses induced by oral EYHBV-STF solution were compared with those by EYHBV-STF injection and by different dosages (injection and oral) of porcine spleen HBV-specific transfer factor (PSHBV-STF), porcine spleen nonspecific transfer factor, and egg yolk extracts from non-immunized hens.

RESULTSThe prepared EYHBV-STF oral solution, which met the standards for biological products, could inhibit leukocyte adhesion in vitro and significantly enhance mouse foot pad swelling, demonstrating its capability of transferring antigen-specific delayed type hypersensitivity reactions to naive recipient. EYHBV-STF oral solution also significantly improved the immune organ index in mice (P<0 01) with similar effects to those caused by EYHBV-STF injections and by PSHBV-STF injection and oral solution.

CONCLUSIONOrally administered EYHBV-STF and EYHBV-STF injection both possess hepatitis B antigen-specific cellular immune activity and can significantly enhance specific cellular immune responses.

Animals ; Chickens ; Egg Yolk ; chemistry ; Hepatitis B ; drug therapy ; Hepatitis B Antigens ; Hepatitis B virus ; drug effects ; Immunity, Cellular ; Immunization ; Mice ; Swine ; Transfer Factor ; administration & dosage ; pharmacology

Glycerol monolaurate (GML) has been widely used as an effective antibacterial emulsifier in the food industry. A total of 360 44-week-old Hy-Line brown laying hens were randomly distributed into four groups each with six replicates of 15 birds, and fed with corn-soybean-meal-based diets supplemented with 0, 0.15, 0.30, and 0.45 g/kg GML, respectively. Our results showed that 0.15, 0.30, and 0.45 g/kg GML treatments significantly decreased feed conversion ratios (FCRs) by 2.65%, 7.08%, and 3.54%, respectively, and significantly increased the laying rates and average egg weights. For egg quality, GML drastically increased albumen height and Haugh units, and enhanced yolk color. Notably, GML increased the concentrations of polyunsaturated and monounsaturated fatty acids and reduced the concentration of total saturated fatty acids in the yolk. The albumen composition was also significantly modified, with an increase of 1.02% in total protein content, and increased contents of His (4.55%) and Glu (2.02%) under the 0.30 g/kg GML treatment. Additionally, GML treatments had positive effects on the lipid metabolism of laying hens, including lowering the serum triglyceride and total cholesterol levels and reducing fat deposition in abdominal adipose tissue. Intestinal morphology was also improved by GML treatment, with increased villus length and villus height to crypt depth ratio. Our data demonstrated that GML supplementation of laying hens could have beneficial effects on both their productivity and physiological properties, which indicates the potential application of GML as a functional feed additive and gives us a new insight into this traditional food additive.
Albumins/analysis* ; Animals ; Chickens ; Diet ; Dietary Supplements ; Egg Yolk/chemistry* ; Female ; Gonadal Steroid Hormones/blood* ; Intestines/cytology* ; Laurates/administration & dosage* ; Lipid Metabolism ; Monoglycerides/administration & dosage* ; Oviposition/drug effects* ; Ovum ; Oxidative Stress

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