Immunoglobulin IgY Research Articles

Review Article: Chicken Immunoglobulin IgY Preparation, and Its Applications in Prevention and / or Control of Some Microbial Affections.

Review Article: Chicken Immunoglobulin IgY Preparation, and Its Applications in Prevention and / or Control of Some Microbial Affections.

Inhibition of sexual stage-specific proteins results in reduced numbers of sexual stages and oocysts of Cystoisospora suis (Apicomplexa: Coccidia) in vitro

Parasites of the order Coccidia (phylum: Alveolata, subphylum: Apicomplexa) have sophisticated life cycles that include a switch from asexual to sexual development, characterised by distinct cell types. During the development of gametes (gamogony), substantial changes occur at the cellular and subcellular levels, leading to cell fusion of micro- and microgametes, and the development of a zygote that forms a protective outer layer for environmental survival as an oocyst, the transmissible stage. Studies on the porcine coccidian Cystoisospora suis already identified changes in transcription profiles during different time points in the parasite’s development and identified proteins with potential roles in the sexual development of this parasite. Here, we focus on three proteins that are possibly involved in the sexual development of C. suis. Enkurin and hapless protein 2 (HAP2) play important roles in signal transduction and gamete fusion during the fertilisation process, and oocyst wall forming protein 1 (OWP1) is a homologue of oocyst wall forming proteins of related parasites. We evaluated their locations in the different life cycle stages of C. suis and their inhibition by specific antibodies in vitro. Immunolocalization detected enkurin in merozoites and sporulated oocysts, HAP2 in merozoites and microgamonts, and OWP2 in merozoites, macrogamonts, oocysts and sporozoites. Up to 100% inhibition of the development of sexual stages and oocyst formation with purified chicken immunoglobulin IgY sera against recombinant enkurin, HAP2, and especially OWP1, were demonstrated. We conclude that the three investigated sexual stage-specific proteins constitute targets for in vivo intervention strategies to interrupt parasite development and transmission to susceptible hosts.

Influence of designer diet and holy basil (Ocimum sanctum) leaves on yolk colour, yolk carotenoid, and immunity status of layers

A biological experiment of six weeks duration, followed by several laboratory investigations was carried out to study the effect of designer layer mash (DLM) containing full fat flaxseed, oil rich sardine fish, holy basil leaf meal (BLM), vitamin E and organic selenium (Sel-plex), on yolk colour, carotenoid and immune status of layers. Both designer diet and BLM had significantly improved the yolk colour and pigment levels. They also exhibited a synergistic effect. The DLM had significantly increased the vitamin E and selenium levels in the egg; but BLM had no such effect. The immune status of the layers, expressed as Immunoglobulin IgY and HI titre, indicated that both designer diet as well as the BLM had significantly improved the IgY levels and HI titre.

Monoclonal antibodies against loggerhead sea turtle, Caretta caretta, IgY isoforms reveal differential contributions to antibody titers and relatedness among other sea turtles

Serum from loggerhead sea turtles, Caretta caretta, was collected from the southeast Atlantic Ocean during routine summer monitoring studies in 2017. Serum immunoglobulin IgY was purified and used to develop IgY isoform-specific monoclonal antibodies (mAb). mAb LH12 was developed against the 66 kDa heavy chain of IgY, mAb LH1 was developed against the truncated heavy chain of approximately 37 kDA, and mAb LH9 was developed against the 23 kDa light chains. mAb LH9 reacts with the light chains of all sea turtles, mAb LH12 reacts with the long heavy chain of all sea turtles within the family Cheloniidae, and mAb LH1 reacts with the truncated form of IgY in both olive and Kemp's ridley turtles. Circulating IgY antibodies against three different marine bacterial pathogens were determined in 16 loggerhead samples using these mAbs. mAb LH12 detects higher titers than mAb LH1, and mAb LH9 detects the highest titers.

Assessment of persistent organic pollutants and their relationship with immunoglobulins in blood of penguin colonies from Antarctica

ABSTRACT: Persistent organic pollutants (POPs) may affect the immune system of seabirds, and some field studies have examined this effect. There are evidences that POPs cause negative effects on the immune system, but in Antarctic penguins data are scarce. In order to assess the risks that POPs may have on wild animals, some immunological studies constitute a good alternative. Determining the levels of immunoglobulins can deliver relevant information about the potential impact of POPs in seabirds from different environments. However, there are very few records in penguins so far. The aim of the present work was to quantify the concentrations of polychlorinated biphenyls (PCBs), hexachlorobenzene (HCB), dichlorodiphenyltrichloroethane (LDDT), endosulfans, and immunoglobulin IgY in blood samples of colonies of chinstrap penguins (Pygoscelis antarctica) that inhabit in the Antarctic Peninsula area. Analysis of POPs in blood samples was carried out through extraction with QuEChERS method, followed by gas chromatography. Detection of IgY concentrations was achieved by ELISA test, using anti-chicken antibodies. There was some significant correlation between immunoglobulin and the presence of PCBs and endosulfans. Even though the levels of these pollutants are low to cause any biological effect on the birds, the results are indicating some influence on the IgY concentrations in penguin blood.

Impact give Specific immunoglobulin IgY isolated from the egg yolk in different ways in the productive performance of Ross 308 of experimental infected with Eimeria tenella parasite

The current study was conducted in the laboratory of the Faculty of Science / University of Anbar, for the period from 09/27/2012 until 11/17/2012 day for the purpose of comparison methods give treat disease Coccidosis in the productive performance of broiler through the use of 135 chick is one day old unsexal ( Ross strain 308) where distributed randomly to nine treatment and by five replicates per treatment and 3 chicks each repeater (15 chicks / treatment), gavage chicks in treatment first with distilled water (control negative) , and treatment second chicks for by gavage (Intra - Crop) challenge dose for the E. tenella locally isolated and by 5x 104 Oocyst / chick (2.5 ml) at 28 days for the treatment of positive control For the purpose of comparison methods give immunoglobulin IgY locally isolate, has been giving him a concentration of 11.013748 mg / ml amount of 0.5 ml / chick on all transactions, and in the third treatment given by antibodies capsule was given in the fourth treatment by gavage (Intra - Crop) after administration of 1 ml of acidity equalizer after preparing a tea spoon of equivalent acidity in 10 ml of distilled water (Sodium bicarbonate 2.80 gm, Tartaric acid 1.33gm, Citric acid 0.82 gm, Flavour to 5gm) . In the fifth treatment injected antibodies peritoneal membrane, and the sixth-treatment antibody was injected in the muscle (thigh, chest, neck) and in the seventh treatment injected objects counterrevolution in the yolk sac one day old and in the eighth treatment antibody injected under the skin (thigh, chest, neck) and in the ninth treatment antibody was injected intravenously. Notes from the current results of the experiment that the antibody injection in peritoneal membrane has led to a significant increase in body weight and the weighted increase in the seventh week of life compared to the rest of the transactions. In addition, got a significant increase (p> 0.05) in cumulative feed conversion ratio while significantly decreased the proportion of cumulative mortality (1-49 days) in the treatment of peritoneal injection and yolk sac injection treatment compared to the rest of the treatment. Inferred from the current study that giving intravenous IgY peritoneal membrane was significantly better than the rest of the transactions in its effect on the productive performance down Representative proportion of cumulative mortality and improve feed conversion efficiency, which is reflected in the body weight and increase the cumulative weighted.

IgY Antibody: A Promising Diagnostic and Therapeutic Tool

Immunoglobulin IgY, which is produced by chicken, reptiles and amphibians, has attracted the interest ominously in scientific community due to its unique properties and wider applicability. Moreover, IgY production in chicken favours reduced use of lab animals and is in complete concordance to animal welfare. Consequently, in this review, we aim to provide a brief overview about structural and biochemical properties of IgY, as well as advantages of its usage as diagnostic and therapeutic or prophylactic reagents.

Use specific immunoglobulin IgY purified from egg yolk and see its effect on the weights and lengths of the internal organs in chickens infected parasite Eimeria tenella

Been obtained for the isolation of a local parasite Eimeria tenella by taking the cecca of 850 samples from slaughterhouses chickens in the city of Ramadi, and after purification and In vitro Sporolated diagnosed through the shape and size of the parasite and the infection, as has been confirmed diagnosis of the sample in the veterinary hospital / Ramadi , he received the strain challenge by strengthening the isolation and written consent of Chicken type Ross 308 five times, has been cracking the parasite by an ultrasound Sonicater and extract Sporocyst and Sporozoite in fourth Dose , injected Challenge Strain then in laying hens Type Luhman in different areas under the skin and muscle of the chest and thigh muscle and the muscle of the neck after mixing with the solution of Freund's incomplete oil adjuvant, was then the collection of eggs from the first day of the first dose until the end of the fourth dose of the control sequence to increase immunoglobulin IgY in egg yolk, extracted immunoglobulin IgY from egg yolk on a daily basis and has Dialysis and the deportation of protein extract crude by Vertical Electrophoresis with some standard protein, and measuring the concentration of crude protein by means of a spectroscope optical Spectrophotometer at wavelength 540, depending on Baurit it reached its highest level 137.12604 mg/egg , were separated IgY using Technology Column Chromatography packed gel Sephacryl S300 HR 95 cm high and 1.6 cm diameter and Determenation of molecular weight by separating the dye Blue Dextran 2000 and seven proteins that record different molecular weights and the work of the curve index, were also measured concentration of IgY extracted after separation column by Spectrophotometer it reached its highest level 117.61764 mg/egg, was deported IgY by Electrophoresis with standard IgG for the detection of a package Kama Globulin , conducted revealed clumping of IgY with strain fierce by the use of Single radial immunodiffusion test and Double immunodiffusion Ouchterlony test to determine the extent of specialization against the type of isolation and extraction of focus appropriate for use as a treatment in subsequent experiments, you saved the sample IgY by freezing degree zero -4 C The results showed significant differences between the treatments especially in infection region ( ceacum ) and that the best way to cure by giving injection therapy in the peritoneum and egg yolk inside body compared with negative control and the rest of the transactions .

CORAL SNAKE ANTIVENOM PRODUCED IN CHICKENS (Gallus domesticus)

The production of anti-snake venom from large mammal's blood has been found to be low-yielding and arduous, consequently, antivenom immunoglobulins for treatment are achieved regularly as polyvalent serum. We have standardized an undemanding technique for making purified immunoglobulin IgY antivenom consisting of polyclonal antibodies against coral snake venom in the egg yolk of immunized hens. We have adapted a reported process of antibody purification from egg yolks, and achieved 90% antibody purity. The customized technique consisted of the removal of lipids from distilled water-diluted egg yolks by a freeze–thaw sequence. The specific immunoglobulins were present in the egg yolk for up to 180 days postimmunization. Therefore, by means of small venom quantities, a significant amount of immunoglobulins were found in an adequately purified state (The obtained material contained about 90% pure IgY). The antigen binding of the immunoglobulins was detected by a double immunodiffusion test. Titers of antibodies in the yolk were estimated with a serum protection assay (Median effective dose = ED50) (ED50= 477 mg/kg). Given that breeding hens is economically feasible, egg gathering is noninvasive and the purification of IgY antibodies is quick and easy, chicken immunization is an excellent alternative for the production of polyclonal antibodies. To the best of our knowledge, this is the first coral snake antivenom prepared in birds.

Peptides accompanying chicken egg yolk IgY – alternative methods of isolation and immunoregulatory activity

In the hen immune system the egg content plays as significant a role in the development of the chick as colostrum does in newborn mammals. One of the most important proteins in this system seems to be the main yolk immunoglobulin IgY. It has been shown that IgY is accompanied by an immunostimulatory polypeptide complex named yolkin. In this report the biological activities of yolkin separated by means of four different procedures are presented. It was shown that yolkin acts as an inducer rather than a modulator of cytokine and nitric oxide release, and does not participate in the protection of cells against destructive effects of reactive oxygen species. However, using the perchloric acid procedure it is possible to obtain a peptide fraction with higher inducing activity, stronger antioxidant properties and ability to decrease the NO level induced by lipopolysaccharide. The results obtained show that it is feasible to select one of the presented methods of yolkin isolation that yields a product of particular activity. The properties of yolk peptides not only indicate their roles in the development of chicks, but can also be useful for the regulation of some immunological disturbances.

Electrochemical immunosensor based on ensemble of nanoelectrodes for immunoglobulin IgY detection: Application to identify hen's egg yolk in tempera paintings

A nanostructured electrochemical biosensor for detecting proteins of interest in work of art, in particular in tempera paintings, is presented. To determine egg yolk we focus here on the determination of immunoglobulin IgY. The transducers are nanoelectrode ensembles (NEEs), prepared via membrane templated electroless deposition of gold. Because of their geometrical and diffusion characteristics, NEEs are characterized by significantly low detection limits, moreover they display the capability of capturing proteins by interaction with the polycarbonate membrane of the NEE. At first, the proteic component of the paint is extracted by ultrasonication in an aqueous buffer, then IgY is captured by incubation on the NEE. The immunoglobulin is detected by treatment with anti-IgY labeled with horse radish peroxidase (Anti-IgY-HRP). The binding of the Anti-IgY-HRP is detected by recording the electrocatalytic signal caused by addition of H2O2 and methylene blue. The sensor detection capabilities are tested by analyzing both paint models, prepared in the lab, and real samples, from paintings of the XVIII–XX century. Multivariate exploratory analysis is applied to classify the voltammetric patterns, confirming the capability to differentiate egg-yolk tempera from other kind of tempera binders as well as from acrylic or oil paints.

Development of goose monoclonal IgY antibodies for use as a passive therapeutic. (113.6)

Abstract When vaccination is not cost effective or possible, one of the primary alternatives is passive immunotherapies. Passive therapeutic antibodies have been shown to be effective in the treatment, not only of infectious agents, but also for toxins, venoms, and cancer targets. Recent research in our laboratory as well as others, has demonstrated the potential of avian-derived passive therapeutic antibodies. We have demonstrated goose-derived polyclonal antibodies, from either sera or eggs, are therapeutic for several viral diseases. The avian immunoglobulin IgY is the primary avian antibody isotype. IgY is similar to mammalian IgG, but lacks the ability to interact with mammalian Fc receptors, complement, and other factors that promote inflammation. IgY’s potential as a therapeutic agent is boosted by its higher avidity than IgG and recognizes more epitopes than mammalian antibodies. These advantages are due to the evolutionary distance between mammalian and avian species. In this work we present initial studies to develop goose mIgY, utilizing traditional mammalian monoclonal protocol that will be commercialized against viral or other targets responsive to single epitope treatment, with the product displaying the inherent advantages of avian antibodies.

Evaluation the effect of the use Immunoglobulin IgY immunity of infected chickens with the Eimeria tenella parasite

Been obtained for the isolation of local Eimeria tenella by taking the Cecca of 850 samples from chickens slaughterhouses in the city of Ramadi, and after purification was diagnosed through the shape and size of the parasite and the infection , obtained the challenge strain by strengthening the isolation in Chicken type Ross 308 five times, injected challenge strain in laying hens Type Luhman under the skin and muscle of the chest and thigh after mixing with Freund's incomplete oil adjuvant solution, then get IgY from egg yolk, which began collected three days after the third dose, detection of IgY by using singular and double immunodiffusion test, use the IgY against Eimeria tenella in the treatment concentration of 1/4 with a dose of 5x 104 of the challenge strain and three replicates of treatment, showed the experiment results of the treated chicks with IgY, three days after giving the challenge dose has given the proportion of immunity amounted to 51.12%, while the results given the challenge dose with a solution of IgY given the proportion of immunity amounted to 75.55%, and finally give the challenge dose after giving the solution IgY in three days gave the proportion of immunity amounted 88.88% without any damage in chicks treated in the experiment compared with the control treatment of positive and which gave the proportion of immunity amounted 95.55% compared to control negative, which gave the proportion of immunity amounted 17.78%, as demonstrated by the study of statistical significant differences at the level of probability of 0.05 for all transactions except for the first treatment with the treatment of five .

A new purification process for goose immunoglobulin IgY(ΔFc) with hydrophobic charge-induction chromatography

Hydrophobic charge-induction chromatography (HCIC) is a novel bioseparation technique, especially for antibody purification. In this study, HCIC was used for the purification of goose immunoglobulin IgY(ΔFc). IgY(ΔFc) is a kind of unique immunoglobulin existing in waterfowl, which natively lacks C H3 and C H4 domains of heavy chains and has some particular therapeutic applications. A new HCIC ligand, 2-mercapto-1-methyl-imidazole (MMI), was coupled to the divinyl-sulfone-activated agarose matrix to prepare the HCIC gel. The adsorption isotherms of IgY(ΔFc) were investigated at different pHs. The saturated capacity of IgY(ΔFc) at pH 5.0 could reach 187.5 mg/ml gel, and the pH-dependent adsorption behaviors were found. After the goose plasma was pre-treated with caprylic acid to precipitate some impurities, the supernatant could be directly loaded onto the HCIC column for IgY(ΔFc) separation. The operation conditions were optimized, including the loading pH, elution pH and the loading volume. High separation efficiency was obtained with final purity of 98.6% and the yield of 85.0%. The results indicate that the new process with HCIC can be a promising technology for the cost-effective separation of IgY(ΔFc) from plasma.

Anti-enteropathogenic Escherichia coli immunoglobulin Y isolated from eggs laid by immunised Leghorn chickens

IgY, the egg yolk immunoglobulin, equivalent to the IgG from mammals, has been used in veterinary practice for passive immunisation against bacterial or viral infectious diseases. Enteropathogenic Escherichia coli (EPEC) is the main etiological agent of infantile diarrhoea in Brazil and other developing countries. Our aims were to isolate immunoglobulin IgY from egg yolk laid by EPEC -immunised Leghorn chickens and to study its reactivity to the antigens from this pathogen, including some virulence factors. Leghorn chickens were immunised with a bacterial suspension intramuscularly (three hens) or intravenously (three hens) or with PBS (two hens). Eggs were collected over a period of 17 weeks. IgY isolation procedures were carried out by salt precipitation (ammonium sulphate, in solid form) followed by centrifugations and dialysis. Final preparations were submitted to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS - PAGE), enzyme-linked immunosorbent assay (ELISA) and immunoblotting. All immunised animals developed good levels of antibodies reactive to whole bacteria or lipopolysaccharide (LPS), in contrast to the control ones. Immunoblottings allowed the recognition of several antigenic fractions of bacterial antigens, some of which had a molecular weight compatible with bacterial virulence factors, confirming the efficacy of the immunisation and the adequacy of the method.

Randomized, Placebo‐Controlled, Clinical Trial of Hyperimmunized Chicken Egg Yolk Immunoglobulin in Children With Rotavirus Diarrhea

ABSTRACTBackgroundHyperimmunized bovine colostrum containing antibodies has been shown to be effective in the treatment of rotavirus diarrhea. Antibodies derived from eggs of immunized hens may be a less expensive and more practical alternative. In this study, children with proven rotavirus diarrhea were treated with immunoglobulin extracted from eggs of chicken immunized with human rotavirus strains.MethodsIn a randomized, double‐blind study, 79 children with known rotavirus diarrhea were assigned to receive either 10 g hyperimmune egg yolk (HEY) daily in four equally divided doses for 4 days (HEY group) or a similar preparation obtained from nonimmunized chicken (placebo group). The daily stool frequency and amount, oral rehydration solution (ORS) intake, and presence of rotavirus in the stool were monitored for 4 days.ResultsIn the HEY‐treated group. there was significant reduction in stool output (in grams per kilogram per day; HEY vs. placebo; 87 ± 59 vs. 120 ± 75, P = 0.03), and significant reduction of ORS intake (in milliliters per kilogram per day) (HEY vs. placebo; 84 ± 46 vs. 122 ± 72, P = 0.008) on day 1 and clearance of virus on day 4 (HEY vs. placebo; 73% vs. 46%, P = 0.02). There was, however, no difference in diarrheal duration between the groups.ConclusionsTreatment with HEY against four human rotavirus strains resulted in modest improvement of diarrhea associated with earlier clearance of rotavirus from stools. These results indicate an encouraging role of HEY in the treatment of rotavirus‐induced diarrhea in children. Further studies are needed to optimize the dose and neutralization titer and thus improve the efficacy of egg yolk immunoglobulin IgY derived from immunized hens.

Application of an ELISA-elution assay as a screening tool for dissociation of yolk antibody–antigen complexes

A modified enzyme-linked immunosorbent assay termed ELISA-elution assay was used as a screening tool to compare the efficiency of eluents for the dissociation of hen yolk immunoglobulin IgY–bovine IgG complexes. The potential denaturing effects of the eluents were also monitored. Different buffers (pH 2.3–7.5), containing various types and concentrations of salts (NaCl, (NH 4) 2SO 4 and MgCl 2) as well as polyols (ethylene glycol (EG) and glycerol) were compared to the commonly reported glycine·HCl (pH 2.8) buffer and to a commercially available eluent, Actisep™. Acidic pH buffers, Actisep™ and MgCl 2 (3.5 M with EG or 4 M without EG) all successfully dissociated IgY from immobilized IgG. However, some denaturation was apparent using MgCl 2 and, to a lesser extent, Actisep™. Furthermore, these same eluents demonstrated a diminished ability for liberating IgG from immobilized IgY IgG. Information on eluent efficacy obtained by the ELISA-elution assays was applied to selectively isolate lower affinity antibodies for immunoaffinity column chromatography.