Saline environments are enriched with halophilic archaea and number of virus-like particles. Viruses infecting halophilic archaea are still the worst studied group of viruses. The aim of our studies was to elucidate the infection of Haloarcula hispanica with various morphology viruses and to perform electrochemical on-line monitoring for viral infection analysis. Culture-dependent approaches were used to study Haloarcula hispanica infection with various morphology halophilic viruses: SH1, HHIV2, HHPV1, His1, His2, HHTV1 and HHTV2. Electrochemical measurements were performed to evaluate the changes in membrane voltage (ΔΨ), the efficiency of infected cells respiration and changes in membrane permeability.
For evaluation of the influence of the metabolic poisons to the Har. hispanica growth rate KCN, NaN3 and NaAs were used. The results of experiments showed that the inhibitors of respiration repressed the growth. 10 mM of KCN blocked the growth totally, while Har. hispanica growed slowly in 40 mM of NaN3.
Various morphology viral infection studies showed that the multiplication of archaeal viruses is effectiveness on soft medium then in a liquid one. SH1 virus titer on soft medium - 4,5*1011 pfu/ml liquid - 3,7*1011 pfu/ml; HHPV1: on soft medium - 1,7*1011 pfu/ml, in liquid - 4,0*1010 pfu/ml; His2 – 1,2*1012 pfu/ml and 2,3*1011 pfu/ml. We determined the adsorption rates of SH1, HHIV2, His1 and His2 viruses too. Halovirus adsorption rate constants varied from 4,5*10-10 to 2,9*10-13 ml/min and the influence of virus morphology on the adsorption rate was negligible. From one-step experiments we found out that the life cycle of halophilic viruses infecting Har. hispanica continues 3 – 10 h depending on the virus. The entry mechanism of virus SH1 differs from His2. SH1 depolarised the membrane of infected cells. At the same time His2 did not cause any changes in depolarisation or permeabilization.
Raktažodžiai: Haloarcula hispanica, haloviruses, viral infection