Evaluation of high hydrostatic pressure effect on human adenovirus using molecular methods and cell culture

Abstract

Human adenoviruses (HAdV) are shed in human faeces and can consequently contaminate environmental waters and possibly be transferred to foods by irrigation. Therefore, efficient inactivation technologies for water and foods are needed. High hydrostatic pressure (HHP) processing is a non-thermal, energy-efficient and rapid emergent inactivation technology, which has been widely studied to eliminate pathogenic microorganisms in foods. We have applied HHP to HAdV-2 in water and cell culture medium (CCM) and measured the effect on virus infectivity and genome and capsid integrity, by using infectivity assay, real-time PCR (qPCR) and qPCR with prior enzymatic treatment (ET-qPCR) with Proteinase K and DNase I. While lower pressures did not provide satisfactory inactivation levels, 400 and 600 MPa treatments were estimated to reduce virus infectivity by approximately 6 log₁₀ units when effectively applied for 93s and 4s, respectively (i.e., excluding come up times of the pressure unit). However, virus genome remained intact even when higher pressures were applied. While acidic pH protected HAdV-2 from inactivation with HHP, no baroprotective effect was observed when 1% sucrose was added to the CCM. On the other hand, 10 mM CaCl₂ added to the CCM was estimated to protect HAdV-2 from HHP with longer treatment times (>10 min). When virus was treated in bottled mineral water, significantly higher infectivity reduction was observed compared to the same treatment in CCM. In conclusion, HHP was shown to effectively reduce HAdV-2 infectivity up to 6.5 log₁₀ units within 4s and can thus contribute to public health protection for food- and water-borne virus transmission. However, its precise effect is matrix dependent and therefore matrix-specific evaluations need to be considered for assuring reliable inactivation in practice.