Abstract. Between July and October 1996, a West Nile (WN) fever epidemic occurred in the southern plain and Danube Valley of Romania and in the capital city of Bucharest, resulting in hundreds of neurologic cases and 17 fatalities. In early October 1996, entomologic and avian investigations of the epidemic were conducted in the city of Bucharest and nearby rural areas. Thirty (41%) of 73 domestic fowl sampled had neutralizing antibody to WN virus, including 5 of 13 ducks (38%), 1 of 1 goose, 19 of 52 chickens (37%), 1 of 1 peahen, and 4 of 6 turkeys (67%). Seroprevalence in domestic fowl (27%, or 7 of 26) from the urban Bucharest site was not significantly different (P ϭ 0.08, by Fisher's exact test) than rates at three rural sites (50%, or 23 of 46). Serum collected from one of 12 Passeriformes, an Erithacus rubecula, was positive for neutralizing antibody to WN virus. A total of 5,577 mosquitoes representing seven taxa were collected. Culex pipiens pipiens accounted for 96% of the mosquitoes collected. A single virus isolate, RO97-50, was obtained from a pool of 30 Cx. p. pipiens females aspirated from the walls and ceiling of a blockhouse located near the center of Bucharest, resulting in a minimum infection rate of 0.19 per 1,000. Antisera prepared against RO97-50 failed to distinguish among RO97-50, WN virus strain Eg101, and Kunjin (KUN) virus strain MRM16. A 2,323-basepair DNA fragment of the envelope (E) glycoprotein gene from RO97-50 and a Romanian WN virus strain obtained from a human cerebrospinal fluid sample, RO96-1030, were sequenced. Phylogenetic analyses of 23 WN virus strains and one KUN virus strain using the amino acid and nucleotide sequences for a small portion of the E gene suggest the existence of two large lineages of viruses. Bootstrap analysis of the nucleotide alignment indicated strong support (95%) for a lineage composed of WN virus strains from northern Africa, including isolates from Egypt and Algeria, and west, central, and east Africa, all of the European isolates, those from France and Romania, an Israeli isolate, and an isolate of KUN virus from Australia. The nucleotide sequence of RO97-50 was identical to the sequence of a WN virus isolate obtained from Cx. neavei mosquitoes from Senegal and Cx.

In response to the 1996 West Nile (WN) fever epidemic that occurred in Bucharest and southeastern Romania, a surveillance program was established. The surveillance system detected 39 clinical human WN fever cases during the period 1997-2000: 14 cases in 1997, 5 cases in 1998, 7 cases in 1999, and 13 cases in 2000. Thirty-eight of the 39 case-patients lived in the greater Danube Valley of southern Romania, and 1 case-patient resided in the district of Vaslui, located on the Moldavian plateau. The estimated annual case incidence rate for the surveillance area during the period 1997-2000 was 0.95 cases per million residents. Thirty-four cases were serologically confirmed, and 5 cases were classified as probable. Twenty-four case-patients presented with clinical symptoms of meningitis (62%), 12 with meningoencephalitis (31%), 1 with encephalitis (3%), and 2 with febrile exanthema (5%). Five of the 39 cases were fatal (13%). Fourteen case-patients resided in rural areas, and 25 in urban and suburban areas, including 7 case-patients who resided in Bucharest. The ages of case-patients ranged from 8 to 76 years with a median age of 45 years. Twenty-four case-patients were males and 15 were females. Dates of onset of illness occurred from May 24 through September 25, with 82% of onset dates occurring in August and September. Limited entomological surveillance failed to detect WN virus. Retrospective sampling of domestic fowl in the vicinity of case-patient residences during the years 1997-2000 demonstrated seroprevalence rates of 7.8%-29%. Limited wild bird surveillance demonstrated seroprevalence rates of 5%-8%. The surveillance data suggest that WN virus persists focally for several years in poorly understood transmission cycles after sporadic introductions or that WN virus is introduced into Romania at relatively high rates, and persists seasonally in small foci.

Background Depending on geographic location, causes of encephalitis, meningoencephalitis and meningitis vary substantially. We aimed to identify the most frequent causes, clinical presentation and long-term outcome of encephalitis, meningoencephalitis and meningitis cases treated in the Inselspital University Hospital Bern, Switzerland. Methods In this monocentric, observational study, we performed a retrospective review of clinical patient records for all patients treated within a 3-year period. Patients were contacted for a telephone follow-up interview and to fill out questionnaires, especially related to disturbances of sleep and wakefulness. Results We included 258 patients with the following conditions: encephalitis (18%), nonbacterial meningoencephalitis (42%), nonbacterial meningitis (27%) and bacterial meningoencephalitis/meningitis (13%). Herpes simplex virus (HSV) was the most common cause of encephalitis (18%); tick-borne encephalitis virus (TBEV) was the most common cause of nonbacterial meningoencephalitis (46%), enterovirus was the most common cause of nonbacterial meningitis (21%) and Streptococcus pneumoniae was the most common cause of bacterial meningoencephalitis/meningitis (49%). Overall, 35% patients remained without a known cause. After a median time of 16 months, 162 patients participated in the follow-up interview; 56% reported suffering from neurological long-term sequelae such as fatigue and/or excessive daytime sleepiness (34%), cognitive impairment and memory deficits (22%), headache (14%) and epileptic seizures (11%). Conclusions In the Bern region, Switzerland, TBEV was the overall most frequently detected infectious cause, with a clinical manifestation of meningoencephalitis in the majority of cases. Long-term neurological sequelae, most importantly cognitive impairment, fatigue and headache, were frequently self-reported not only in encephalitis and meningoencephalitis survivors but also in viral meningitis survivors up to 40 months after acute infection.

Background Depending on geographic location causes of encephalitis, meningoencephalitis and meningitis vary substantially. We aimed to identify most frequent causes, clinical presentation as well as long-term outcome of encephalitis, meningoencephalitis and meningitis cases treated in the Inselspital, University Hospital Bern, Switzerland. Methods In this monocentric, observational retro- and prospective cohort study, we performed a retrospective review of clinical patient records for all patients treated during 3 years. Patients were contacted prospectively for a telephone follow-up interview and to fill out questionnaires, especially related disturbances of sleep and wakefulness. Results We included 258 patients: encephalitis (18%), non-bacterial meningoencephalitis (42%), non-bacterial meningitis (27%) and bacterial meningoencephalitis/meningitis (13%). Herpes simplex virus (HSV) was the most frequent cause of encephalitis (18%), tick borne encephalitis virus (TBEV) of non-bacterial meningoencephalitis (46%), enterovirus of non-bacterial meningitis (21%) and Streptococcus pneumoniae of bacterial meningoencephalitis/meningitis (49%). Overall, 35% patients remained without known cause. After a median time of 16 months, 162 patients participated in the follow-up interview, thereof 56% indicated to suffer from neurological long-term sequels such as fatigue and/or excessive daytime sleepiness (34%), cognitive impairment and memory deficits (22%), headache (14%) and epileptic seizures (11%). Conclusions In the largest tertiary care University hospital in Switzerland TBEV was the overall most frequently detected infectious cause, with a clinical manifestation of meningoencephalitis in the majority of cases. Long-term neurological sequels, most importantly cognitive impairment, fatigue and headache were frequently self-reported not only in encephalitis and meningoencephalitis but also viral meningitis survivors up to 40 months after the acute infection.