Specific Chicken Egg Yolk Antibodies against Enterotoxigenic Escherichia coli K99 in Serum and Egg Yolk of Immunized Laying Hens

Subject Areas : Camel

ا.  کرم زاده-دهاقانی ¹ (Department of Animal Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran)
آ.  توحیدی ² (Department of Animal Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran)
م.  ژندی ³ (Department of Animal Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran)
ن.  مژگانی ⁴ (Department of Biotechnology, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran)

Received: 2019-04-23 Accepted : 2019-08-31 Published : 2020-03-01

Keywords: Passive immunity, chicken antibodies, <i>E. coli</i>, IgY,

Abstract :

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 suspension 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&rsquo;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 assay (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 &lt;0.05). The level of binding activity of specific IgY in serum and EY increased in immunized hens (p &lt;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 antibodies could inhibit E. coli K99 in vitro growth.

References:

Amro W.A., Al-Qaisi W. and Al-Razem F. (2018). Production and purification of IgY antibodies from chicken egg yolk. J. Genet. Eng. Biotechnol. 16, 99-103. 

Bi Y., Yang C., Diao Q. and Tu Y. (2017). Effects of dietary supplementation with two alternatives to antibiotics on intestinal microbiota of preweaned calves challenged with Escherichia coli K99. Nature. 7, 5439-5443.

Bradford M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254.

Cai Y.C., Guo J., Chen S.H., Tian L.G., Steinmann P. and Chen M.X. (2012). Chicken egg yolk antibodies (IgY) for detecting circulating antigens of Schistosoma japonicum. Parasitol. Int. 61, 385-390.

Diraviyam T., Zhao B., Wang Y., Schade R., Michael A. and Zhang X. (2014). Effect of chicken egg tolk antibodies (IgY) against diarrhea in domesticated animals: A systematic review and meta-analysis. PLoS. One. 9, e97716.

Hashish E.A., El Damaty H.M., Tartor Y.H. and Abdelaal A.M. (2016). Epidemiological study of diarrheagenic Escherichia coli virulence genes in newborn calves. Pakistan Vet. J. 36(1), 54-58.

Hatta H., Tsuda K., Akachi S., Kim M., Yamamoto T. and Ebina T. (1993). Oral passive immunization effect of anti Human rotavirus IgY and its behavior against proteolytic enzymes. Biosci. Biotechnol. Biochem. 57, 1077-1081.

He J., Wang Y., Sun S. and Zhang X. (2015). Evaluation of chicken IgY generated against Canine parvovirus viral-like particles and development of enzyme-linked immunosorbent assay and immunochromatographic assay for Canine parvovirus detection. Viral. Immuol. 28, 489-494.

Jiang Z., Jiang X., Li C., Xue H. and Zhang X. (2016). Development of an IgY antibody-based immunoassay for the screening of the CYP2E1 inhibitor/enhancer from herbal medicines. Front. Pharmacol. 7, 502-510.

Kubo R., Zimmerman B. and Grey H. (1973). Phylogeny of immunoglobulins. Pp. in 417-477 in The Antigens.M. Sela, Ed. Academic Press, New York.

Kuby J. (1997). Overview of the immune system. J. Immuol. 3, 3-24.

Lee E., Sunwoo H., Menninen K. and Sim J. (2002). In vitro studies of chicken egg yolk antibody (IgY) against Salmonella enteritidis and Salmonella typhimurium. Poult. Sci. 81, 632-641.

Li C., Ren H., Schade R. and Zhang X. (2017). A novel and efficient immunoglobulin Y extraction method using poloxamer-polyethylene glycol. Prep. Biochem. Biotechnol. 47, 739-743.

Li X., Jing K., Wang X., Li Y. and Zhang M. (2016). Protective effects of chicken egg yolk antibody (IgY) against experimental Vibrio splendidus infection in the sea cucumber (Apostichopus japonicus). Fish Shellfish Immunol. 48, 105-111.

Li X., Nakano T., Sunwoo H., Paek B., Chae H. and Sim J. (1998). Effects of egg and yolk weights on yolk antibody (IgY) production in laying chickens. Poult. Sci. 77, 266-270.

Liou J., Lin Y., Lien T., Shiau J., Tailiu J. and Tai C. (2010). Production of egg yolk immunoglobulin against Escherichia coli from white Leghorn and Lohmann layer. J. Anim. Vet. Adv. 10(18), 2349-2356.

Mahdavi A., Rahmani H., Nili N., Samie A. and Soleimanian-Zad S. (2010). Chicken egg yolk antibody (IgY) powder against Escherichia coli O78: K80. J. Anim. Vet. Adv. 9, 366-373.

Owusu-Asiedu A., Nyachoti C. and Marquardt R. (2003). Response of early-weaned pigs to an enterotoxigenic Escherichia coli (K88) challenge when fed diets containing spray-dried porcine plasma or pea protein isolate plus egg yolk antibody, zinc oxide, fumaric acid, or antibiotic. J. Anim. Sci. 81(7), 1790-1798.

Pereira E.P.V., Van-Tilburg M.F., Florean E.O.P.T. and Guedes M.I.F. (2019). Egg yolk antibodies (IgY) and their applications in human and veterinary health: A review. Int. Immunopharmacol. 73, 293-303.

Razali N.M. and Wah Y.B.J. (2011). Power comparisons of shapiro-wilk, kolmogorov-smirnov, lilliefors and anderson-darling tests. J. Stat. Model. Anal. 2, 21-33.

Reddy P., Ramlal S., Sripathy M.H. and Batra H.V.J. (2014). Development and evaluation of IgY immunocapture PCR ELISA for detection of Staphylococcus aureus enterotoxin A devoid of protein A interference. J. Immunol. Methods. 408, 114-122.

Sădziene A., Rosa P., Thompson P., Hogan D. and Barbour A.J. (1992). Antibody-resistant mutants of Borrelia burgdorferi: In vitro selection and characterization. J. Exp. Med. 176, 799-809.

SAS Institute. (2004). SAS^®/STAT Software, Release 9.4. SAS Institute, Inc., Cary, NC. USA.

Schade R., Calzado E.G., Sarmiento R., Chacana P.A., Porankiewicz-Asplund J. and Terzolo H.R. (2005). Chicken egg yolk antibodies (IgY-technology): A review of progress in production and use in research and human and veterinary medicine. Altern. Lab. Anim. 33, 129-154.

Shahrani M., Dehkordi F.S. and Momtaz H. (2014). Characterization of Escherichia coli virulence genes, pathotypes and antibiotic resistance properties in diarrheic calves in Iran. Biol. Res. 47, 28-39.

Sun S., Mo W., Ji Y. and Liu S.J.R. (2001). Preparation and mass spectrometric study of egg yolk antibody (IgY) against Rabies virus. Rapid Commun. Mass Spectrom. 15, 708-712.

Sunwoo H., Lee E., Menninen K., Suresh M. and Sim J.J. (2002). Growth inhibitory effect of chicken egg yolk antibody (IgY) on Escherichia coli O157: H7. J. Food Sci. 67, 1486-1494.

Vega C., Bok M., Chacana P., Saif L., Fernandez F. and Parreno V. (2011). Egg yolk IgY: protection against rotavirus induced diarrhea and modulatory effect on the systemic and mucosal antibody responses in newborn calves. Vet. Immunol. Immunopathol. 142(3), 156-169.

Weber K. and Osborn M. (1969). The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J. Biol. Chem. 244, 4406-4412.

Zhang X., Diraviyam T., Li X., Yao G. and Michael A. (2016). Preparation of chicken IgY against recombinant E2 protein of bovine viral diarrhea virus (BVDV) and development of ELISA and ICA for BVDV detection. Biosci. Biotechnol. Biochem. 80, 2467-2472.

Zhen Y.H., Jin L.J., Guo J., Li X.Y., Lu Y.N. and Chen J. (2008). Characterization of specific egg yolk immunoglobulin (IgY) against mastitis-causing Escherichia coli. Vet. Microbiol. 130, 126-133.