Quist-Rybachuk G V, Nauwynck H J, Kalmar I D
Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium; Veterinary R&D - Veos, Akkerstraat 4A, B-8750 Zwevezele, Belgium.
Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
Vet Microbiol. 2015 Dec 31;181(3-4):283-8. doi: 10.1016/j.vetmic.2015.10.010. Epub 2015 Oct 22.
Emergence of porcine epidemic diarrhea virus (PEDV) resulted in massive neonatal mortality in the North-American and Asian pork industry. Measures to prevent its geographical spread are of utmost importance to safeguard susceptible porcine populations. The major infection route is direct or indirect faecal-oral contact. Adequate biosafety measures should be in place at all levels of the swine production chain, including feed and feed ingredients. Present study aimed to investigate the sensitivity of PEDV to thermal inactivation at neutral and alkaline pH in presence or absence of porcine plasma. Cell culture medium and porcine plasma at different pH (7.2, 9.2, 10.2) and temperature conditions (4 °C, 40 °C, 44 °C, 48 °C) were inoculated to a final titer of 5.5 log10 TCID50 PEDV/ml, incubated for up to 120 min and the residual infectivity was determined by endpoint dilution assay. Irrespective of presence of plasma, PEDV was not sensitive to pH 7.2-10.2 at 4 °C. At moderate temperatures (≥40 °C), both alkaline pH and presence of plasma potentiated thermal inactivation. Inactivation of 8 log10 TCID50/ml plasma within 30 min (8D value<30 min) by moderate pH and temperature would denote potential industrial processing conditions that ensure safety towards PEDV while limiting denaturation of bioactive components. Virus-spiked plasma required heat treatment of 40 °C and alkalinization to pH 9.2 to achieve 8 log10 reduction within such time. At pH 10.2 and 48 °C, the 8D value was 4.6 min in plasma and 15.2 min in MEM. Here we propose heat-alkalinity-time (HAT) pasteurization as a highly efficient method to inactivate PEDV during industrial processing of porcine plasma.
猪流行性腹泻病毒(PEDV)的出现导致北美和亚洲养猪业大量新生仔猪死亡。采取措施防止其地理扩散对于保护易感猪群至关重要。主要感染途径是直接或间接的粪口接触。在养猪生产链的各个层面,包括饲料和饲料原料,都应采取适当的生物安全措施。本研究旨在调查在有或没有猪血浆存在的情况下,PEDV在中性和碱性pH值下对热灭活的敏感性。将不同pH值(7.2、9.2、10.2)和温度条件(4℃、40℃、44℃、48℃)下的细胞培养基和猪血浆接种至最终滴度为5.5 log10 TCID50 PEDV/ml,孵育长达120分钟,并通过终点稀释法测定残余感染性。无论血浆是否存在,PEDV在4℃时对pH 7.2 - 10.2均不敏感。在中等温度(≥40℃)下,碱性pH值和血浆的存在均会增强热灭活作用。通过适度的pH值和温度在30分钟内使8 log10 TCID50/ml血浆失活(8D值<30分钟),将意味着潜在的工业加工条件既能确保对PEDV的安全性,又能限制生物活性成分的变性。加毒血浆需要在40℃进行热处理并碱化至pH 9.2才能在该时间内实现8 log10的降低。在pH 10.2和48℃时,血浆中的8D值为4.6分钟,MEM中的8D值为15.2分钟。在此,我们提出热 - 碱 - 时间(HAT)巴氏灭菌法作为一种高效方法,用于在猪血浆的工业加工过程中灭活PEDV。