Yolk IgG Research Articles

Effect of dietary manipulation and vaccination of turkey breeder hens on immunoglobulin levels of yolk, yolk sac and neonate poults.

Two hundred turkey breeder hens and 24 viable toms of 30-35weeks age of small white variety were distributed into two treatment groups having four replicates of 25 hens and three toms in each treatment. First four replicates were offered a turkey breeder diet (Diet A) (Nutrient requirements of poultry, 1994, National Academic Press, Washington, DC) and the rest four replicates were maintained on a higher plane of nutrition (Diet B) for 8-week duration. After 6weeks of experimental feeding, two replicates from each treatment groups were vaccinated with ND (R2 B) vaccine. Yolk sac of embryo from birds fed Diet B had a significantly higher (p<.05) IgG, IgM level and HI titre (log 2) than those fed Diet A. HI titre values of embryonic yolk sac from the vaccinated birds fed Diet B were significantly higher (p<.05) than that of the control groups. In addition, HI titre values were significantly higher (p<.05) in the day-old poults of the birds fed Diet B than that of those fed Diet A. There was significantly (p<.01) positive correlation between serum IgG and IgM of the breeder birds and day-old chicks. Similarly, there was significantly (p<.05) positive correlation between yolk IgG and IgM after 1-month experimental feeding and yolk sac IgG and IgM. Positive correlation (p<.05) also existed between yolk sac IgM and day-old chick serum IgM. Furthermore, the HI titres of breeder birds' serum at 14days post-vaccination were positively correlated with their egg yolk after 10 and 15days post-vaccination, yolk sac and day-old chicks. Thus, the study envisaged that a higher immunity in neonate poults from turkey breeders maintained on a higher plane of nutrition may be elicited as there was maternal transfer of antibodies from the serum of breeder birds to their offsprings through their yolk sac.

Immunization of chickens with Salmonella enterica subspecies enterica serovar Enteritidis pathogenicity island-2 proteins

Several structural components of the type III secretion systems (T3SS) encoded by Salmonella pathogenicity island (SPI)-1 and SPI-2 are exposed to the host's immune system prior to/during the infection/invasion process, making them potential vaccine candidates. In this study we evaluated whether chickens vaccinated with SPI-2 T3SS components could mount a significant humoral immune response (as measured by serum IgG titres) and whether these antibodies could be transferred to progeny (as measured by egg yolk IgG titres), and whether vaccinates and progeny of vaccinates could be protected against challenge with SE. The results of our studies show that vaccinated chickens do produce high levels of SPI-2 T3SS specific serum IgG that they are able to transfer to their progeny. It was demonstrated that vaccinates and progeny of vaccinates had lower overall countable recovered Salmonella enterica subspecies enterica serovar Enteritidis (SE) per bird in most situations.

Purificação de IgG de codorna a partir de soro e gema de ovos

The purification of Japanese quail IgG from serum was performed using caprilic acid and ammonium sulfate, and from egg yolk using PEG-6000 and ethanol. After confirming the purification and the concentration of IgG, the yolk samples had twice the amount of protein compared to serum samples. The IgG extracts were analyzed by SDS-PAGE and western blot showing similar results as the ones of chicken IgG used as the standard. So, these methodologies can be used for purifying quail serum or egg yolk IgG, which would enable the development of diagnostic assays.

Structural analysist of N-glycans from gull egg white glycoproteins and egg yolk IgG

We previously showed that the expression of (Gal alpha 1-4Gal)-bearing glycoproteins among birds is related to their phylogeny. However, precise structures of (Gal alpha 1-4Gal)-containing N-glycans were only known for pigeon egg white glycoproteins and IgG. To compare structural features of (Gal alpha 1-4Gal)-containing N-glycans from other species, we analyzed N-glycans of gull egg white (GEW)-glycoproteins, ovomucoid, and ovotransferrin, and gull egg yolk IgG by HPLC, mass spectrometry (MS), and MS/MS analyses. GEW-glycoproteins included neutral, monosialyl, and disialyl N-glycans, and some of them contained Gal alpha 1-4Gal sequences. Bi-, tri-, and tetra-antennary oligosaccharides that lacked bisecting GlcNAc were the major core structures, and incomplete alpha-galactosylation and sialylation as well as the presence of diLacNAc on the branches generated microheterogeneity of the N-glycan structures. Moreover, unlike pigeon egg white glycoproteins, the major sialylation in GEW-glycoproteins is alpha2,3-, but not alpha2,6-linked sialic acids (NeuAc). In addition to the complex-type oligosaccharide, hybrid-type oligosaccharides that lack bisecting GlcNAc were also abundant in GEW-glycoproteins. Gull egg yolk IgG also contained Gal alpha 1-4Gal beta 1-4GlcNAc beta 1- sequences, but unlike pigeon IgG, no Gal alpha 1-4Gal beta 1-4Gal beta 1-4GlcNAc beta 1- sequence was detected. Bi- and tri-antennary complex-type oligosaccharides with bisecting GlcNAc and with core fucosylation as well as high-mannose-type oligosaccharides were the major structures in gull IgG. Our data indicated that some N-glycans from both GEW-glycoproteins and gull IgG contain the Gal alpha 1-4Gal beta 1-4GlcNAc beta 1- sequence, but the ratio of alpha-Gal-capped residues to non-alpha-Gal-capped residues in the nonreducing termini of N-glycans is much lower than that in those of pigeon glycoproteins.

Transgenerational immunity in a bird–ectoparasite system: do maternally transferred antibodies affect parasite fecundity or the offspring's susceptibility to fleas?

During egg formation, female birds deposit antibodies against parasites and pathogens they were exposed to before egg laying into the yolk. In captive bird species, it has been shown that these maternal immunoglobulins (maternal yolk IgGs) can protect newly hatched offspring against infection. However, direct evidence for such benefits in wild birds is hitherto lacking. We investigated (1) if nestling Great Tits Parus major originating from eggs with naturally high levels of maternal yolk IgG are less susceptible to a common, nest‐based ectoparasite, (2) if maternal yolk IgGs influence nestling development and in particular, their own immune defence, and (3) if there is a negative correlation between levels of maternal yolk IgG in host eggs and the reproductive success of ectoparasitic fleas feeding on the nestlings. Counter to expectations, we found no indication that maternally transferred yolk IgGs have direct beneficial effects on nestling development, nestling immune response or nestling resistance or tolerance to fleas. Furthermore, we found no negative correlation between host yolk IgG levels and parasite fecundity. Thus, whereas previous work has unequivocally shown that prenatal maternal effects play a crucial role in shaping the parasite resistance of nestling birds, our study indicates that other egg components, such as hormones, carotenoids or other immuno‐active substances, which bird females can adjust more quickly than yolk IgG, might mediate these effects.

Effect of bursal anti-steroidogenic peptide and immunoglobulin G on neonatal chicken B-lymphocyte proliferation

In attempts to identify antibodies for Bursal Anti-Steroidogenic Peptide (BASP), rabbit serum was observed to reduce phorbol ester-stimulated chicken B-lymphocyte proliferation comparable to BASP. These experiments investigated the effects of IgG on B-lymphocyte proliferation. In Experiment 1, 3% rabbit serum decreased B-lymphocyte proliferation. In Experiment 2, 2 mg/ml of intact rabbit IgG or 0.65 mg/ml of IgG papain digest products, Fab and Fc, decreased B-lymphocyte proliferation. The combination of BASP and either Fab or Fc was observed to have at least an additive anti-proliferative effect. In Experiment 3, 0.01 mg/ml of either rabbit or chicken IgG, or 1.0 mg/ml of rabbit or 0.01 mg/ml of chicken Fab, Fc, and the pepsin digestion product F(ab′) 2 was observed to have an anti-proliferative effect. No combined effects of BASP and IgG or IgG digest products were observed for this experiment. In Experiment 4, 12 mg/ml of chicken egg yolk IgG or 1.2 mg/ml Fab was found to suppress B-lymphocyte proliferation. Additionally, an additive effect of 12 mg/ml of IgG with BASP was again observed. The present studies suggest that IgG and its digestion products reduce phorbol-stimulated B-lymphocyte proliferation in vitro and combined treatment with IgG and BASP may have at least an additive anti-proliferative effect on B-lymphocyte proliferation.

Development of maternal IgG-free chick obtained from surgically bursectomized hen

IgG-free eggs and chicks were developed, so as to study the role of maternal IgG in the development of the immune system. Surgical bursectomy on the 18th day of incubation deprived chickens of B cells and eliminated IgG synthesis. Bursectomized chickens are usually dead before sexual maturity under conventional conditions. When surgically bursectomized chickens were housed in an isolated clean room and antibiotics were administered to them, they could survive to sexual maturity. Finally, we succeeded in obtaining IgG-free fertilized eggs and maternal IgG-free chicks from surgically bursectomized hens. The amount of yolk IgG in IgG-free eggs was one-ten thousandth less than that in normal eggs. The level of IgM in the serum of maternal IgG-free chicks reached six times higher than that of normal chicks 5 days after hatching.

Serum and Egg Yolk IgG Antibody Titers from Laying Chickens Vaccinated with Pasteurella multocida

Through determining the serum and egg yolk antibody titers in immunized laying hens to Pasteurella multocida regularly, the growth-decline trend of the egg yolk antibody levels was found to be similar to that of the serum antibody levels (r = 0.94), but the growth and decline of the egg yolk antibody seemed to be delayed 3-6 days compared with that of the serum antibody, and the egg yolk antibody titers were generally lower than those of the serum antibody (P < 0.01). Serum and egg yolk antibody levels declined 3 and 6 days, respectively, after booster immunizations. The higher the antibody levels were before booster immunization, the more they declined.

An improved chromatographic method for the separation of egg yolk IgG into subpopulations utilizing immobilized metal ion (Fe 3+) affinity chromatography

An improved methodology is described for the separation of yolk IgG into subpopulations using immobilized metal ion (Fe 3+) affinity chromatography. The yolk IgG was first extracted using a prechilled, pre-acidified method. After extraction, the yolk IgG was then fractionated using an Fe 3+ column. Using an ascending pH gradient, four IgG containing peaks were well resolved based upon the elution pH, specific activity and the relative avidity index.

Development of Preventive Measures Based on the Aetiology of Dental Caries: A Review

Mutans streptococci including Streptococcus mutans and Streptococcus sobrinus are aetiologically associated with the development of dental caries in humans and experimental animals. These organisms produce glucosyltransferases (GTases), which catalyse the synthesis of adherent glucan from sucrose, promoting the adherence of the organisms to the tooth surface. In addition, they release large quantities of acids from various dietary sugars. These are the essential virulence factors of mutans streptococci. We found that structural isomers of sucrose such as palatinose and trehalulose were not utilised by the organisms and no significant glucans or acids were produced. Another interesting sucrose substitute would be sugar alcohol; maltitol and palatinit are good examples. Other unique compounds developed recently are oligosaccharides rich in isomaltose and panose, which contain α (1→6) glucosidic linkage. They strongly inhibit glucan synthesis from sucrose by GTases, and cannot be degraded by most oral streptococci. These sucrose substitutes are now manufactured on an industrial scale. Enzymatic inhibition of GTases is an alternative measure to suppress caries development. Polymeric polyphenol compounds from oolong tea and hen egg yolk IgG specific for cell-associated GTase of mutans streptococci have been shown to exhibit marked GTase inhibition and decrease in caries development in rats. Keywords: dental caries, mutans streptococci, glucosyltransferase, palatinose, polyphenol, maltitol, passive immunity.

A model for the prediction of relative titres of avian malaria and Aspergillus spp. IgG in Jackass penguin ( Spheniscus demersus) females based on maternal IgG in egg-yolk

A model for the prediction of IgG titres in females of the Jackass penguin ( Spheniscus demersus) was developed, based on IgG which was maternally tranemitted to the yolk of urembryonated eggs produced by these females. However, prediction of the female titre based on the titre of embryonated eggs may be inadequate. Blood samples from 10 S. demersus females and their corresponding embryonated ( n=10) and unembryonated ( n=49) eggs were analysed by indirect ELISA for avian malaria ( Plasmodium relictum, P. elongatum) IgG and Aspergillus spp. IgG. There was no correlation between humoral responses to avian malaria and Aspergillus spp. in females or in their eggs. Avian malaria and Aspergillus spp. titres were significantly higher ( P<0.05) in the eggs than in the corresponding females and were significantly correlated ( P<0.01) with the blood titres, r=0.84, r=0.89, respectively. No correlation was found between titres of embryo yolk-sac (embryonated eggs) and the blood of their female parent; however, the embryo blood and the corresponding female titres were significantly ( P<0.05) correlated (avian malaria, r=0.74; Aspergillus spp., r=0.69). Blood and the corresponding egg-yolk (unembryonated eggs) IgG titres regressed significantly ( P<0.01). Female IgG titre ( y) is related to (unembryonated) egg-yolk IgG titre by the significant ( P<0.05) regression y=−0.61+1.46 X for avian malaria, and y=−0.02+0.85 X for Aspergillus spp., with ±95% prediction limits of ±0.15 and ±0.12, respectively. This model provides access to serological information on the remote free-ranging Jackass penguins and captive Jackass-peaguin colonies without the necessity of stressful blood collection.

A Two-Step Procedure for Purification of Hen Egg Yolk Immunoglobulin G: Utilization of Hydroxypropylmethylcellulose Phthalate and Synthetic Affinity Ligand Gel (Avid AL®)

A method for isolation of egg yolk IgG was performed by subjecting egg yolks to the delipidizing action of 5% hydroxypropylmethyl-cellulose phthalate (HPMCP) solution after dilutions of egg yolk with distilled water. Eightfold dilution of egg yolk with distilled water mixed with .25% (vol/vol) HPMCP was found to be effective for precipitation of yolk lipoprotein and recovery of Ig-rich supernatant. The lipid content in the supernatant after removal of the resulting precipitate was .4% of egg yolk. Immunoglobulin G remained in this supernatant, and was purified by affinity chromatography using Avid AL® synthetic ligand gel to isolate and purify the IgG. There was a high degree of purity of IgG recovered as assessed by SDS-PAGE and no loss in its biological properties.

Serological Response of Chickens to Infection with Salmonella gallinarum-S. pullorum Detected by Enzyme-Linked Immunosorbent Assay

The serological response to Salmonella pullorum and S. gallinarum infection in chickens was studied with an indirect enzyme-linked immunosorbent assay (ELISA). In broiler chickens, a more virulent strain of S. pullorum produced a significantly lower serum IgG titer than did a less virulent strain. In laying hens, the serum and egg-yolk IgG titers were very similar. In chickens infected with S. gallinarum, high IgG titers persisted for 30 weeks. In chickens reinfected with this strain, each reinfection was followed by transitory increases in IgG lasting no longer than 2 weeks. Serum samples from Brazil taken from a laying flock with evidence of fowl typhoid showed much higher antibody levels than did those from three uninfected flocks. Using lipopolysaccharide as the detecting antigen, infections caused by these salmonellae could be differentiated from those caused by other groups. Incorporation of the appropriate flagella antigen in the ELISA allowed differentiation between infections caused by S. pullorum and S. enteritidis.

Immune responses in chickens against Salmonella typhimurium monitored with egg antibodies

Three mature hens were immunized with an Aro-mutant of Salmonella typhimurium beginning with a subcutaneous dose in adjuvant followed by two oral boosters. Isotypeantibodies were measured in the white and yolk eggs collected weekly over a period of 230 days. Two hens showed a memory response to the first oral boosters, with large increases in egg yolk IgG and smaller increases in IgA and IgM antibodies in egg whites. Smaller amounts of IgA and IgM antbodies were found in egg yolks, and a slight increase in IgG occured in the whites. One hen showed an increase in serum titers of all isotypes against S. typhimurium. The second hen had serum titers before immunization was started which did not change. The third hen had a high level of IgM in the white of eggs before immunization was started. This hen showed erratic responses in egg white antibodies following immunization, no increase in IgA or IgM in yolks and only a slight increase in IgG, no increase in serum IgG, and was the only hen with a high level of IgM antibody against S. typhimurium in the bile, conditions reflecting a state of oral tolerance. With the exception of this hen, the results showed that IgA and IgM antibodies were aroused in hens by immunization with an avirulent mutant of S. typhimurium, and that these antibodies were present in the white of eggs from immunized hens.

Isolation of Egg Yolk Immunoglobulin-Rich Fractions Using Copper-Loaded Metal Chelate Interaction Chromatography

Immunoglobulins were partially purified from egg yolk by metal chelate interaction chromatography (MCIC), after initial preparation of yolk with alginate precipitation. Under the phosphate-buffered equilibration conditions chosen, chicken IgG did bind to the MCIC column when small sample sizes were applied. With increasing amounts applied to the column, IgG was found to be preferentially displaced into the unbound fractions, resulting in an increase in IgG purity in the unbound. By selectively pooling the unbound fractions from a two-thirds copper-saturated 10mL MCIC column, 104mg IgG was obtained from four eggs at 75% purity. For industrial applications, the MCIC method described in the present study appears to have several advantages over current methods of yolk IgG separation.

Serum and egg antibody responses in chickens to Escherichia coli.

The effects of Freund’s adjuvants on antibody production in chickens against E. coli whole cells were examined. The levels of anti-E. coli IgG antibodies in serum were higher when Freund’s complete (FCA) or incomplete adjuvant (FIA) was administered than that without adjuvant. Production of antibodies recognizing E. coli cells and their lipopolysaccharide was enhanced by FIA, while both FIA and FCA enhanced production of antibodies recognizing outer membrane components. In contrast, serum IgM antibody levels were higher when no adjuvant was used. Anti-E. coli IgG antibodies in serum were efficiently transferred to egg yolk, giving antibody activity in egg yolk similar to that in serum. However, anti-E. coli IgM antibodies were not detected in the egg, suggesting that egg (white) IgM was not influenced by antigenic stimulation of the humoral immune system. Antimicrobial activity of the egg yolk IgG was highest when the bacteria antigen was injected with FIA.

Eggs: Conveniently packaged antibodies. Methods for purification of yolk IgG

Eggs from immunized chickens may provide a convenient and inexpensive source of antibodies. We describe two simple and efficient methods for purification of IgG from yolk. The antibody is rendered useful for most currently employed immunological techniques. Amounts of antibody corresponding to almost half a litre of antiserum may be recovered from a chicken in one month.

IgG receptors on foetal chick yolk sac.

In the chicken, maternal IgG is deposited in the oocyte yolk and subsequently transported intact by the foetal chick yolk sac into the chick circulation. Such transport appears to require a specific membrane-localized IgG receptor. Using the assay of 125I-yolk IgG binding, we have investigated the existence and properties of such receptors in yolk sac tissue disks. We find that: (1) There are specific, 125I-IgG binding sites on yolk sac tissue; (2) specific yolk 125I-IgG binding has a distinct pH optimum at pH 6.0--6.3; (3) the apparent equilibrium dissociation constant of specific 125I-IgG binding is 2--3 X 10(-6) M; and (4) heterologous proteins compete less efficently than yolk IgG for 125I-IgG binding.