Gujral Naiyana, Yoo Heejoo, Bamdad Fatemeh, Lee Kwang Y, Suh Joo-Won, Sunwoo Hoon
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, 114 St-89 Ave., Edmonton, Alberta, Canada.
Center for Nutraceutical and Pharmaceutical Materials, Myongji University, Yongin, Gyeonggi, 449-728, Korea.
Curr Pharm Biotechnol. 2017;18(5):400-409. doi: 10.2174/1389201018666170425120036.
Enterotoxigenic Escherichia coli (ETEC) is the main cause of fatal diarrhea in piglets during the first week of life and over the time of weaning. Pathogenesis of ETEC-causing diarrhea involves intestinal colonization mediated by fimbriae. Although, both IgY and egg yolk phosvitin (PV) possess antimicrobial activity, their combined activity has not been explored. A combination of IgY specific for ETEC and metal-chelating PV may show synergistic effect in reducing the growth of ETEC by inhibiting bacterial proliferation and stipulating protection against ETEC infection.
The goal of this study was to determine the effects of anti-ETEC IgY and PV on in vitro growth inhibition of ETEC strains possessing K88 and K99 fimbriae prevalent in the porcine population.
Anti-K88 and -K99 IgY antibodies were obtained from egg yolks of 23-week-old Single- Comb White Leghorn hens immunized with K88 and K99 fimbriae of ETEC, respectively, with high titres sustained over 6 to 8 weeks of the immunization period. Specific IgY, PV, and PV-hydrolysate from alcalase-hydrolysis under high hydrostatic pressure (PVH-Alc-HHP) alone or in combination, were used to treat ETEC K88 and K99 cultures at optimal concentrations of 100 μg/mL, 1 mg/mL, and 1 mg/mL, respectively, for 24 h.
PVH-Alc-HHP demonstrated the highest degree of hydrolysis, 38.9%. Combined use of IgY and PVH-Alc-HHP showed the highest bactericidal effect resulting in ETEC K88 and K99 growth inhibition of 2.8 and 2.67 log CFU/mL, respectively.
Combined IgY-PVH effectively control ETEC, therefore holds a great potential for microbial control in veterinary pharmaceutical industry.
产肠毒素大肠杆菌(ETEC)是仔猪出生后第一周及断奶期间致命性腹泻的主要病因。ETEC引起腹泻的发病机制涉及菌毛介导的肠道定植。虽然,卵黄免疫球蛋白(IgY)和卵黄磷蛋白(PV)均具有抗菌活性,但它们的联合活性尚未得到探索。针对ETEC的特异性IgY与金属螯合PV的组合可能通过抑制细菌增殖和提供针对ETEC感染的保护作用,在减少ETEC生长方面显示出协同效应。
本研究的目的是确定抗ETEC IgY和PV对具有猪群中普遍存在的K88和K99菌毛的ETEC菌株的体外生长抑制作用。
分别从用ETEC的K88和K99菌毛免疫的23周龄单冠白来航母鸡的蛋黄中获得抗K88和-K99 IgY抗体,在免疫期的6至8周内维持高滴度。单独或组合使用特异性IgY、PV以及在高静水压下经碱性蛋白酶水解得到的PV水解产物(PVH-Alc-HHP),分别以100 μg/mL、1 mg/mL和1 mg/mL的最佳浓度处理ETEC K88和K99培养物24小时。
PVH-Alc-HHP表现出最高的水解程度,为38.9%。IgY和PVH-Alc-HHP联合使用显示出最高的杀菌效果,导致ETEC K88和K99的生长抑制分别为2.8和2.67 log CFU/mL。
IgY-PVH组合能有效控制ETEC,因此在兽药行业的微生物控制方面具有巨大潜力。