Isolation of infectious Lloviu virus from Schreiber’s bats in Hungary

Some filoviruses can be transmitted to humans by zoonotic spillover events from their natural host and filovirus outbreaks have occured with increasing frequency in the last years. The filovirus Lloviu virus (LLOV), was identified in 2002 in Schreiber’s bats (Miniopterus schreibersii) in Spain and was subsequently detected in bats in Hungary. Here we isolate infectious LLOV from the blood of a live sampled Schreiber’s bat in Hungary. The isolate is subsequently sequenced and cultured in the Miniopterus sp. kidney cell line SuBK12-08. It is furthermore able to infect monkey and human cells, suggesting that LLOV might have spillover potential. A multi-year surveillance of LLOV in bats in Hungary detects LLOV RNA in both deceased and live animals as well as in coupled ectoparasites from the families Nycteribiidae and Ixodidae. This correlates with LLOV seropositivity in sampled Schreiber’s bats. Our data support the role of bats, specifically Miniopterus schreibersii as hosts for LLOV in Europe. We suggest that bat-associated parasites might play a role in the natural ecology of filoviruses in temperate climate regions compared to filoviruses in the tropics.

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No statistical test was used to determine sample size. Sample sizes were variable between sampling events, depending on the actual colony size and the availability of animals. During in vitro laboratory experiments, the LLOV positive sample number limited the number of experiment replicates.
No data were excluded from the study Depending on the available amount of bat sera different measurement and dilution strategies were applied: measured as duplicate with repeated experiment -bat1, 169,170,99,115,130; measured in duplicate with single experiment -bat2,102,138; measured in single in one experiment -bat98,110,118,143.
Randomization is not relevant in our study, since we sampled wild animals. This sampling is affected by multiple random factors, there is no option for randomized sampling. Animal activity, number of animals for sampling and accessibility of animals are not predictable. We sampled all animals which could be catched.
Blinding is not relevant in our study, since we sampled wild animals and performed screening on site. This sampling is affected by multiple random factors, there is no option for blinded sampling. Animal activity, number of animals for sampling and accessibility of animals are not predictable. Circumstances are not applicable for blinded sampling. Study design and relevance do not requires blinded sampling.