Emoke Ferenczi

Emergence of Usutu Virus in Hungary

ABSTRACT In 2001, Usutu virus (USUV), a mosquito-borne flavivirus of the Japanese encephalitis virus serogroup related to West Nile virus and previously restricted to sub-Saharan Africa, emerged in wild and zoo birds in and around Vienna, Austria. In order to monitor the spread of the infection, a dead bird surveillance program was established in Austria and in neighboring Hungary. In Hungary, 332 dead birds belonging to 52 species were tested for USUV infection between 2003 and 2006. In the first 2 years, all birds investigated were negative. In August 2005, however, USUV was detected in organ samples of a blackbird (Turdus merula), which was found dead in Budapest, Hungary, by reverse transcription-PCR, immunohistochemistry, and in situ hybridization. In July and August 2006, a further six dead blackbirds tested positive for USUV, and the virus was isolated from organ samples of one bird. These birds were also found in urban areas of Budapest. The nearly complete genomic sequence of one Hungarian USUV strain was determined; it was found to share 99.9% identity with the strain that has been circulating in Austria since 2001. This result indicates that the USUV strain responsible for the blackbird die-off in Budapest most likely spread from Austria to Hungary instead of being independently introduced from Africa.

Tick-borne encephalitis outbreak in Hungary due to consumption of raw goat milk

A tick-borne encephalitis outbreak involving 25 patients of 154 exposed persons occurred in Hungary in August 2007. None of the patients had a history of tick-bite, however all of them drank unpasteurized raw goat milk from the same farm. The aim of this study was to identify the goats on the farm which could have spread the infection through their milk. Blood samples were taken from 75 goats on the farm and were examined by various serological methods, namely indirect immunofluorescent assay, hemagglutination inhibition, microneutralization and an ELISA adapted to testing material from goats, to determine antibody levels in the serum. The four methods have proved different levels of specificity. The least specific was the indirect immunofluorescent assay, which showed a low titre in all sera. Comparison of the results of the other three methods indicates that two sera were positive for anti-TBEV IgG and one for anti-TBEV IgM. The goat with the IgM positive serum sample could have been a source of the infected milk. It has been concluded that serological results for goats by the different methods should be compared before final diagnosis because the specificity of methods in use can differ significantly.

Co-circulation of three pathogenic hantaviruses: Puumala, Dobrava, and Saaremaa in Hungary.

Hantaviruses (Bunyaviridae) cause hemorrhagic fever with renal syndrome (HFRS) in Eurasia and hantavirus (cardio)pulmonary syndrome (HCPS) in the Americas. HFRS is caused by Hantaan virus (HTNV), Seoul virus (SEOV), Dobrava virus (DOBV), Saaremaa virus (SAAV), and Puumala virus (PUUV). Of those, only HTNV is not present in Europe. In recent years, hantaviruses, described in other parts of Europe, were also detected at various locations in Hungary. To study the genetic properties of Hungarian hantaviruses in detail, sequences of the viral S and M segments were recovered from bank voles (Myodes glareolus), yellow‐necked mice (Apodemus flavicollis), and striped field mice (Apodemus agrarius) trapped in the Transdanubian region. As expected, the sequences recovered belonged, respectively, to PUUV (two strains), DOBV (one strain), and SAAV (one strain). On phylogenetic trees two new Hungarian PUUV strains located within the well‐ supported Alpe‐Adrian (ALAD) genetic lineage that included also Austrian, Slovenian, and Croatian strains. Analysis of the Hungarian SAAV and DOBV genetic variants showed host‐specific clustering and also geographical clustering within each of these hantavirus species. Hungarian SAAV and DOBV strains were related most closely to strains from Slovenia (Prekmurje region). This study confirms that multiple hantaviruses can co‐circulate in the same locality and can be maintained side‐by‐side in different rodent species. J. Med. Virol. 81:2045–2052, 2009.

Severe tick-borne encephalitis in a patient previously infected by West Nile virus.

We describe severe tick-borne encephalitis (TBE) in a patient who had previously experienced West Nile fever, another flavivirus infection endemic in Hungary. Previous West Nile virus infection does not develop immunity either against TBE virus infection or the disease, and it does not mitigate its clinical course. The possibility of antibody-dependent enhancement is considered.

Serologic evidence of West Nile virus infection in patients with exanthema in Hungary.

The presence of WNV in Europe has been well known for decades, although the first human infections and avian outbreaks were diagnosed in Hungary only in 2003. An annual average of 6-8 cases of the neuroinvasive form of WNV infection has been detected in the region since then, but a higher number (17) of WNV associated neuroinvasive disease occurred in 2008. In 2004, a surveillance system was established for monitoring WNV-associated meningo-encephalitis cases in Hungary, but a milder type of illness (with fever, rash and/or influenza like symptoms) is not followed. Fifty-two sera of 45 patients with mild clinical symptoms (fever, exanthema) were tested for anti-WNV antibodies in 2008 in a retrospective study by immunofluorescence test and ELISA. Seven patients had antibodies against WNV, serologic evidence of recent WNV infection was found in 4 out of the 7 patients. Infections could be acquired predominantly in August and in September, which seems to be a risk period for WNV in Hungary. The possibility of a recent WNV infection should be taken into consideration in the occurrence of fever and rush at late summer. Differential diagnosis of exanthematous patients should include WNV serology tests and should be done routinely.