Enterotoxigenic Escherichia coli K99 is one of the dominant pathogens associated with diarrhea of calves. Immunoglobulin Y (IgY), has been used as an inexpensive alternative to antibiotics for the prevention and therapy of several bacterial infections. The study aimed to prepare IgY antibodies against E. coli K99 and to investigate its in vitro effectiveness. E. coli K99 was grown in the tryptic soy broth, and the bacterial sus-pension was inactivated by formaldehyde. Thirty White Leghorn hens allocated to control and treatment groups. 1 mL of the prepared bacterial suspension or sterilized physiological serum emulsified with Freund’ s complete adjuvant was injected at two weeks interval to the hens in treated or control group. The total IgY was purified with polyethylene glycol (PEG) 6000 from EGG YOLKs (EY). Purified IgY fractions were processed for protein concentrations by Bradford assay. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to analyze purified IgY fractions. An indirect enzyme-linked immunosorbent as-say (ELISA) method was used to measure the specific IgY titers in serum and EY. The ability of non-specific and anti-E. coli K99 IgY antibody at 100 and 200 mg/mL concentration was evaluated by growth inhibition assay in vitro. According to the ELISA data, total IgY concentration in serum and EY of control was relatively constant, while increased in the treatment group (P<0. 05). The level of binding activity of specific IgY in serum and EY increased in immunized hens (P<0. 05). Specific IgY had the highest activity for bacterial inhibiting growth at the level of 200 mg/mL. These results suggested the prepared IgY antibod-ies could inhibit E. coli K99 in vitro growth.

The aim of the present study was to investigate effect of additive genetic on EGG weight, YOLK weight and YOLK fatty acid content and estimation of heritability for these traits. In this research, 150 white Japanese quails (50 males and 100 females) were used as the base population (without any pedigree information). In the F2 generation, 100 female quails were randomly selected at the fifth week of age and transferred to laying cages. The traits included EGG weight, YOLK weight and YOLK fatty acid contents. (Co) Variance components and genetic parameters were estimated using multiple animal models by Wombat software. The estimated heritability for EGG weight and YOLK weight were 0.45 and 0.38, respectively. These estimates for YOLK fatty acids ranged from 0.27 (Palmitoleic acid) to 0.45 (Palmetic acid). Genetic correlations between EGG weight and YOLK fatty acids were low, ranging from 0.01 (between EGG weight and Linoleic acid) to 0.10 (between EGG weight and Stearic acid). The higher genetic correlations were between YOLK weight and YOLK fatty acids, varying from 0.10 (between YOLK weight and Linoleic acid) to 0.51 (between YOLK weight and Stearic acid). The results showed that selection for increasing the YOLK weight could lead to producing EGGs with more contents of YOLK fatty acids.

Oxidative damage to membrane lipid is one of the prime events occurring in aging and other undesirable physiological processes. In this study experiments were performed on liposomes (prepared either from crude erythrocyte PHOSPHOLIPIDS or purified EGG YOLK phosphatidylcholine) as models of lipid bilayer portion of biomembranes. The effects of β-carotene, and phospholipid composition on peroxidation process, initiated by Fe2+, were studied. It was found that β-carotene does not show any noticeable antioxidant effect on the peroxidation process initiated by Fe2+ in liposomes prepared from erythrocyte phosphatides, whereas it effectively suppressed the same process in EGG YOLK phosphatidylcholine (EYPC). It is concluded that the anti-/pro-oxidant activity of β–carotene is also dependent on the membrane lipid composition and this may provide an explanation about the conflicting reports on its role in ordinary or promoted oxidation experiments.

This study was conducted to assess the physical and morphological characteristics of fresh, pre-freeze and post-thaw black Bengal buck (Capra hircus) semen processed in Tris-EGG YOLK-citrate-fructose-glycerol (TEYCFG) extender containing 4 different concentrations (2.5, 5, 7.5 and 10%) of EGG-YOLK. Ejaculates were collected once a week for eight weeks from six mature bucks (48 ejaculates) using an artificial vagina. It was found that a general decrease in values of pre-freeze and post-thaw semen parameters such as progressive sperm motility (%), live sperm (%), acrosomal integrity (%) and positive sperm (%) in the hypo-osmotic swelling (HOS) test, but an over-all increase in abnormal spermatozoa compared to fresh semen for extenders containing different concentrations of EGG-YOLK. Best values for all the semen parameters were obtained using extender with 2.5% EGG-YOLK. Of all EGG-YOLK concentrations, the 10% EGG-YOLK supplement caused the highest percentage of abnormal sperm, which was significant (P<0.05), compared to 2.5% and 5% EGG-YOLK containing extenders, but non-significantly (P>0.05) different compared with 7.5% EGG-YOLK containing extender. Only progressive motility was significantly different (P<0.05) within extenders containing 2.5%, 5%, 7.5% and 10% EGG-YOLK. We conclude that Black Bengal buck semen can be cryopreserved effectively with tris-EGG YOLK-citrate-fructose-glycerol extender containing 2.5% EGG YOLK (V/V).

The current study was conducted to evaluate the effect of pigeon plasma EGG YOLK (PPEY) and chicken plasma EGG YOLK (CPEY) compared to combination of PPEY+CPEY in tris-citric acid-fructose diluent to preserve the ram semen quality during liquid-cold storage. Semen samples were collected using the artificial vagina from four Qezel rams twice a week. Then, in case if the samples met the criteria, they were pooled and used for the experiment. Tris-citric acid-fructose based extenders were prepared using PPEY (28%, v/v), CPEY (28%), and the combinations of PPEY (7, 14 and 21%) + CPEY (21, 14 and 7 %), and used for the experiment. Following dilution of samples with extenders, total and forward progressive motility of spermatozoa (evaluated by computer assisted sperm analysis), viability and plasma membrane integrity was assessed at 0, 24, 48 and 72 of after cooling. Furthermore, amounts of malondialdehyde (MDA), as an oxidative indicator, were measured at mentioned time points. Results revealed that forward progressive motility (at 24, 48 and 72 h) and viability (at 72 h) were higher in PPEY (28%) group compared to all combination groups. Moreover, membrane integrity (at 24, 48 and 72 h) was greater in PPEY (28%) relative to combination groups containing 14 and 21% CPEY. Amounts of MDA did not differ among treated groups. In conclusion, combinations of PPEY+CPEY was not effective as PPEY (28%) and CPEY (28%) alone to preservation of ram semen during liquid-cold storage.

Background: Cryptosporidium parvum is a parasitic protozoan that functions as important causative agent of diarrhea in human and animals. The host's immune response to surface antigens of C. parvum has been previously demonstrated. In this respect, the role of humoral immunity in the development of host protective immunity against this protozoon has been well demonstrated. Methods: The effect of specific chicken EGG YOLK antibody (IgY) against recombinant C. parvum P23 was examined. IgY sample was prepared from EGGs of chickens immunized with recombinant C. parvum protein p23 and analyzed with C. parvum lysate and recombinant P23. Results: The anti P23 specific IgY was recognized a protein band with approximately 23 kDa in lysates prepared from the C. parvum oocysts. Also dot blot analysis of recombinant P23 showed that it could be recognized by the anti P23 specific IgY up to 1/1000 dilution of antibody. But the best antibody dilution for immunological studies was determined as 1:200.Conclusion: Since P23 is an immunodominant surface glycoprotein expressed in the early phase of infection, specific IgY against recombinant p23 could be recommended as a favorable candidate for passive immunization against C. parvum infection in human and animals.

Introduction: Chicken is the only avian species in which polyclonal antibodies, like IgG is transported from the hen to the EGG YOLK in a similar manner as the transport of mammalian IgG from the mother to the fetus. Immunoglobulin Y in the chicken is transported to the EGG and accumulates in the EGG YOLK in large quantities. IgY is an EGG YOLK antibody that has been used widely for treatment and prevention of infections in humans and animal. IgY is used for passive protection of the pathogen infections such as Escherichia coli, bovine and human rotavirus, bovine coronavirus, salmonella, staphylococcus and Pseudomonas. IgY is a promising candidate as an alternative to antibiotics. Eschericha coli strains of serotype O157: H7 belongs to a family of pathogenicE. coli called enter hemorrhagic E. coli (EHEC) strains responsible for hemorrhagic colitis, bloody or non-bloody diarrhea, and hemolytic uremic syndrome in humans. This strain of E. coli pathogenises by adhering to host intestinal epithelium and forming bacterial colonies. The purpose of this study was to produce and purify immunoglobulin Y against E. coli O157: H7 and develop specific polyclonal antiE. coli antibody in the EGG YOLK.Materials and Methods: Sixteen-week-old laying hens (Mashhad, Iran) were kept in individual cages with food and water adlibitum. Immunization of hens was performed by intramuscularly injecting killed E. coli O157: H7with an equal volume of Freund’s complete adjuvant into two sides of chest area (Sigma, USA) for the first immunization. Two booster immunizations followed up using complete and incomplete Freund’s adjuvants in two weeks interval. Freund’s adjuvant without antigen was injected to the control group. Two weeks after the last injection, the EGGs were collected daily for eight weeks, marked and stored at 4oC. In order to IgY purification, EGGs were collected. Purification of IgY from EGG YOLK was based on Polson and using PEG6000. Finally, the presence of antibody IgY was confirmed using SDS-PAGE. Purification of IgY was carried out by polyethylene glycolprecipitation method using PEG 6000 powder (Merck, Germany) based on method of Polson. The purified IgY against E. coli was separated using 10% SDS-PAGE. In order to investigate the effect of the specific anti-E. Coli antibody, mice (Razi, Institute of Iran) were randomly distributed into five experimental groups (6mice/group). The mice were kept in conventional animal facilities and received water and food ad libitum. All animal care and procedures were in accordance with institutional policies for animal health and well-being. Experimental groups were including group 1 (mice received IgY orally in drinking water 72 hours before intraperitoneal injection of bacteria and then injected intraperitoneally with 0.5 ml of bacteria E. coli O157: H7), group 2 (mice received IgY orally in drinking water 72 hours before the injection and then injected intraperitoneally with 0.5 ml of deionized water), group 3 (0.5 ml of E. coli O157: H7 incubated with 0.5 ml of the specific anti-E. coli IgY and then 0.5 ml of the incubated solution injected to mice intraperitoneally), group 4 (mice injected with 0.5 ml of IgY) and group 5 (mice received 0.5 ml of E. coli O157: H7).Results and Discussion: We obtained specific EGG YOLK antibody against E. coli O157: H7 by immunizing hens with the killed E. coli O157: H7 antigen. The results showed that the IgY was successfully purified from EGG YOLK. SDS-PAGE analysis showed presence of protein bands 27kDa and 67 kDa of IgY, which correspond to IgY light and heavy chains. Effects of IgY on mice showed that mice received IgY orally in drinking water 72 hours before intraperitoneal injection were protected against bacteria. Also, when specific anti-E. coli IgY was incubated with E.coli O157: H7 for 24 hours and then it was injected to mice led to mice protected against bacteria. The results of our study were agreement with the results of Chae et al (2007). We indicated mice immunized with specific anti-E. coli IgY could be protected against E. coli O157: H7. This phenomenon could be due to specific binding activity of IgY with bacteria that led to the inhibition of bacterial growth E. coli O157: H7.Conclusion: The effectiveness of IgY in suppressing the activity of E. coli O157: H7 was indicated in our study. This could be inferred from the results of the current study that IgY in the EGG YOLK could prevent greater economic losses due to human and animal health from pathogenic bacteria such asE. coli O157: H7. These finding indicated that EGG from immunized hens are potentially useful source of passive immunity.