Insect emergence in relation to floods in wet meadows and swamps in the River Dalälven floodplain.

Annual variation in flood frequency and hydroperiod during the vegetation season has ecological impacts on the floodplain biota. Although many insect groups may have a lower emergence during a flood event, it is poorly known how annual emergence of insects in temporary wetlands is related to the variation in hydrology. Between May and September, we studied the weekly emergence of 18 insect taxa over six consecutive years, 2002-2007, in six temporary flooded wetlands (four wet meadows and two forest swamps) in the River Dalälven floodplains, Central Sweden. We used emergence traps to collect emerging insects from terrestrial and aquatic parts of wet meadows and swamp forests. In all wetlands, the insect fauna was numerically dominated by the orders Diptera, Hymenoptera, Coleoptera and Homoptera. On a weekly basis, 9 out of the 18 insect taxa had lower emergence in weeks with flood than in weeks with no flood, whereas no taxon had a higher emergence in weeks with flood. Over the seasons, we related insect emergence to seasonal flood frequency and length of hydroperiod. The emergence of most studied taxa decreased with increasing hydroperiod, which suggests that emergence after floods do not compensate for the reduced emergence during floods. Only Culicidae and the aquatic Chironomidae sub-families Tanypodinae and Chironominae showed an increase in emergence with increasing hydroperiod, whereas Staphylinidae peaked at intermediate hydroperiod. We conclude that a hydroperiod covering up to 40% of the vegetation season has a significant negative effect on the emergence of most taxa and that only a few taxa occurring in the temporary wetlands are actually favoured by a flood regime with recurrent and unpredictable floods.

The arbovirus vector Culex torrentium is more prevalent than Culex pipiens in northern and central Europe.

Two species of arbovirus vector, Culex torrentium and Culex pipiens (Diptera: Culicidae), occur in several European countries, but difficulties in their accurate identification and discrimination have hampered both detailed and large-scale distribution and abundance studies. Using a molecular identification method, we identified to species 2559 larvae of Cx. pipiens/torrentium collected from 138 sites in 13 European countries ranging from Scandinavia to the Mediterranean coast. In addition, samples of 1712 males of Cx. pipiens/torrentium collected at several sites in the Czech Republic were identified to species based on the morphology of their hypopygia. We found that the two species occur together in large areas of Europe, and that Cx. torrentium dominates in northern Europe and Cx. pipiens dominates south of the Alps. The transition in dominance occurs in central Europe, where both species are roughly equally common. There was a strong correlation between the length of the growing season at different sites and occurrences of the two species. As the growing season increases, the proportion and detection of Cx. torrentium decrease, whereas those of Cx. pipiens increase. The present findings have important consequences for the interpretation of the results of studies on major enzootic and link-vectors of mosquito-borne bird-associated viruses (i.e. Sindbis, West Nile and Usutu viruses), especially in central Europe and Scandinavia.

Francisella tularensis subspecies holarctica occurs in Swedish mosquitoes, persists through the developmental stages of laboratory-infected mosquitoes and is transmissible during blood feeding.

In Sweden, mosquitoes are considered the major vectors of the bacterium Francisella tularensis subsp. holarctica, which causes tularaemia. The aim of this study was to investigate whether mosquitoes acquire the bacterium as aquatic larvae and transmit the disease as adults. Mosquitoes sampled in a Swedish area where tularaemia is endemic (Örebro) were positive for the presence of F. tularensis deoxyribonucleic acid throughout the summer. Presence of the clinically relevant F. tularensis subsp. holarctica was confirmed in 11 out of the 14 mosquito species sampled. Experiments performed using laboratory-reared Aedes aegypti confirmed that F. tularensis subsp. holarctica was transstadially maintained from orally infected larvae to adult mosquitoes and that 25% of the adults exposed as larvae were positive for the presence of F. tularensis-specific sequences for at least 2 weeks. In addition, we found that F. tularensis subsp. holarctica was transmitted to 58% of the adult mosquitoes feeding on diseased mice. In a small-scale in vivo transmission experiment with F. tularensis subsp. holarctica-positive adult mosquitoes and susceptible mice, none of the animals developed tularaemia. However, we confirmed that there was transmission of the bacterium to blood vials by mosquitoes that had been exposed to the bacterium in the larval stage. Taken together, these results provide evidence that mosquitoes play a role in disease transmission in part of Sweden where tularaemia recurs.

A six-year study of insect emergence from temporary flooded wetlands in central Sweden, with and without Bti-based mosquito control.

In temporary wetlands in the River Dalälven floodplains, recurrent but irregular floods induce massive hatching of the flood-water mosquito Aedes sticticus, which causes enormous nuisance. Flood-water mosquito control using the biological larvicide Bacillus thuringiensis var. israelensis (Bti) was commenced in parts of the floodplains during 2002, and here we report the first six years of full-season monitoring of general insect emergence from temporary wetlands with and without treatment. Emergence traps, which were emptied weekly, were used from May to September each year. A total of 137,153 insects of 13 taxonomic orders were collected. Diptera was highly dominating and especially the sub-order Nematocera with 18 families was a very prominent taxon. Bti-treatment effects were analysed by taxonomic order, by sub-order in Diptera and Hemiptera, and by family for Nematocera and Coleoptera for the whole study period. We found no significant negative effects of Bti treatments on the production of insects by taxonomic order, with the exception of Coleoptera in the long term. However, no significant negative effects were found for the Coleoptera families, neither in the short term nor in the long term. There was no significant negative treatment effect on Nematocera production, neither when analyzed for the whole sub-order nor when analyzed by family. However, abundance of Ceratopogonidae was significantly higher in experimental than in reference wetlands. We conclude that Bti-treatment effects on insect production may be minute in comparison to other environmental factors structuring the insect fauna of the temporary wetlands studied.

A sensitive and reliable restriction enzyme assay to distinguish between the mosquitoes Culex torrentium and Culex pipiens.

Culex pipiens pipiens Linnaeus and Culex torrentium Martini (Diptera: Culicidae) are closely related vector species that exist sympatrically in Europe. The two species are morphologically almost identical and can only be distinguished with certainty by characters of the male genitalia. Hence, correct species identification and conclusions on distribution and vector status are very difficult and often neglected. Therefore, we developed a reliable and simple mitochondrial cytochrome c oxidase subunit I (COI) gene restriction enzyme assay to discriminate between Cx. pipiens and Cx. torrentium, based on the analysis of morphologically identified male specimens. We sequenced approximately 830 bp in the 3' region of the mitochondrial COI gene of 18 morphologically identified males of Cx. pipiens and Cx. torrentium. Two restriction enzymes (FspBI and SspI) that could distinguish between the two species according to species-specific differences in these sequences were chosen. The restriction enzymes were tested on 227 samples from Sweden and verified by sequencing 44 of them. The enzyme FspBI correctly identified all investigated samples; the enzyme SspI identified all samples except one Cx. torrentium. We hope the method and the findings presented here will help to shed light on the true distribution and relative proportions of the two species in Europe.

High species richness of Chironomidae (Diptera) in temporary flooded wetlands associated with high species turn-over rates.

Species richness and species turn-over of Chironomidae was studied in irregularly flooded wetlands of the River Dalälven flood-plains in central Sweden. The chironomid fauna, sampled with emergence traps in six wetlands over six summers, contained as much as 135 species, and the cumulative species curves indicated that the regional species pool contain several more species. Recurrent irregular floods may have induced this high chironomid species richness and the high species turn-over in the temporary wetlands, as the dominance between terrestrial and aquatic species shifted between years. Half of the wetlands were treated with Bacillus thuringiensis var. israelensis (Bti) against larvae of the flood-water mosquito Aedes sticticus. These treatments had no significant effect on chironomid species richness, but there was a higher species turn-over between years of primarily low abundance species in the treated wetlands. The cumulative number of species was also higher in the Bti-treated experimental wetlands than in the untreated reference wetlands. Thus, Bti treatment against mosquito larvae seemed to have only small effects on chironomid species richness but seemed to increase the colonisation-extinction dynamics.

Production of wetland Chironomidae (Diptera) and the effects of using Bacillus thuringiensis israelensis for mosquito control.

Massive mosquito nuisance problems, caused by the flood-water mosquito Aedes sticticus, occur after floods in the flood plains of the River Dalälven, central Sweden. Since 2002, the biological mosquito larvicide Bacillus thuringiensis var. israelensis (Bti) has been used to control these mosquitoes. Here, we report results from the first six years of monitoring Chironomidae, the most susceptible non-target organisms, in three wetlands with Bti-treatment against mosquitoes and in three wetlands without treatment. Emergence traps were used for continuous insect sampling from May to September each year, 2002-2007, and were emptied once a week. A total of 21,394 chironomids of 135 species were collected, and the subfamily Orthocladiinae dominated the fauna followed by Chironominae and Tanypodinae. The annual chironomid production in the wetlands was low, with an average of 1917 individuals per m(2), and 42 g ash-free dry weight per m(2). We found no reduced production of chironomids at neither family, nor subfamily level, in Bti-treated as compared to untreated wetlands. This is the first long-term follow-up study of the possible effects of Bti-based mosquito larval control on chironomid species production. In the short-term view, one species had higher production in treated areas. In the long-term view, four species had higher and one species had lower production in treated areas. We conclude that the Bti-based control of flood-water mosquitoes does not cause any major direct negative effects on chironomid production, and therefore does not seem to induce any risk for indirect negative effects on birds, bats or any other predators feeding on chironomids.

Comparison of mosquito (Diptera: Culicidae) populations by wetland type and year in the lower river Dalälven region, Central Sweden.

We studied adult mosquito assemblages in six wetlands, representing three types (wet meadow, alder swamp, and bog), in the lower part of the River Dalälven in Central Sweden during three consecutive years (2000-2002) and evaluated the influence of wetland type and year. Mosquito abundance differed significantly between years but not between wetland types. Mosquito species richness showed no significant variation between years or wetland types. Cluster analysis based on percentage of similarity resulted in three clusters, with high similarity between all wetlands in 2000. Ordination analysis showed that mosquito assemblages were mainly correlated with wetland type and water level increase in the previous month. Hydrological conditions varied between the years and between the wetland types, and our collections also included a year (2000) with extreme flood situations. The floodwater mosquito species Ochlerotatus sticticus was the predominant species with a strong influence on the whole study due to its long-range dispersal ability. The entire region suffered from enormous numbers of Oc. sticticus in 2000. The data from this study provided the basis for the initiation of a mosquito control project in the region.

Different responses of two floodwater mosquito species, Aedes vexans and Ochlerotatus sticticus (Diptera: Culicidae), to larval habitat drying.

Insect larvae that live in temporary ponds must cope with a rapidly diminishing resource. We tested the hypothesis that floodwater mosquitoes would react to diminishing water levels by accelerating larval development time and emerging as smaller adults. Since a reduction in habitat size leads to increased larval densities, we also included two larval densities. Newly-hatched floodwater mosquito larvae, Aedes vexans (87.9% of emerged adults) and Ochlerotatus sticticus (12.0% of emerged adults), were taken from the field and randomly assigned to one of three water level schedules. Survival to adult emergence was significantly affected by the water level schedule. Ae. vexans adults emerged later in the decreasing schedule than the constant water schedule, but time to emergence was not affected by larval density. In the drying water schedule, Ae. vexans adults emerged 6 to 14 days after complete water removal. Adult size was significantly affected by both water level schedule and larval density. Adults of Oc. sticticus emerged earlier in the decreasing than the constant water schedule which was in accordance with our hypothesis, but size was not affected. Our results indicate two different responses of two floodwater mosquito species to diminishing larval habitat. Oc. sticticus accelerated larval development while Ae. vexans larvae showed remarkable survival in humid soil. Both species are often numerous in inundation areas of large rivers, and climatic conditions after a flood might influence which species dominates the adult mosquito fauna.

Biological diversity versus risk for mosquito nuisance and disease transmission in constructed wetlands in southern Sweden.

In southern Sweden, many wetlands have been constructed, and maintaining or increasing biological diversity is often included in the aims. Some wetlands are constructed near human settlements, thus raising the problem of wetlands being associated with mosquitoes (Diptera: Culicidae). Increased biodiversity (including mosquito diversity) is considered desirable, whereas mosquito nuisance from a human point of view is not. Adult mosquito abundance, diversity and species assemblages of constructed wetlands were compared to natural wetlands. The potential of constructed wetlands for mosquito nuisance and transmission of mosquito-borne viruses was evaluated. The study areas included five constructed and four natural wetlands. Mosquito abundance and species richness were higher in the natural than in the constructed wetlands, and showed a positive correlation with wetland size. Mosquito species assemblages formed three clusters, which were not explained by origin, size and water permanence of wetlands. In a redundancy analysis, however, mosquito faunas showed significant relationships with these variables, and size and origin of wetlands were most important. Major nuisance species (multivoltine species feeding on mammals and laying eggs on soil) were found in all wetlands, although in relatively low numbers. Risk assessment for Sindbis virus transmission showed moderate risk for two constructed wetlands near human settlements. It is concluded that small size of constructed wetlands has the advantage of low mosquito numbers from a human point of view. The use of functional groups is recommended as a tool for presenting mosquito data to the public, and for helping communication between scientists and administrative decision makers.

Prevalence of sindbis virus neutralizing antibodies among Swedish passerines indicates that thrushes are the main amplifying hosts.

The Sindbis virus uses birds as vertebrate hosts in the summer amplification cycle, and the virus is transmitted by ornithophilic Culex species. Previous field and experimental studies have shown that mainly passerine birds are involved in the amplification. To delineate the pattern of Sindbis virus infections among passerines, we collected and sampled birds for blood at five study sites located in northern, central, and southern Sweden. All study sites were lowland forested wetlands and humid forests. The blood samples were assayed for Sindbis neutralizing antibodies, and we tested if the prevalence of Sindbis antibodies varied in relation to bird characteristics (i.e., species, body-mass, sex, and age), and environmental factors (i.e., year, month, and location). We found that Sindbis virus infections occurred in almost all passerine species sampled, but that the infection prevalence was unequally distributed among species. The fieldfare, the redwing, and the songthrush each had significantly higher prevalence than the average for all species. Large passerine species had higher infection prevalence than small species. The infection was less prevalent in hatching-year birds than in older birds during June and July, but not in August. Males and females had the same infection prevalence. The prevalence of Sindbis antibodies was higher in central than in southern Sweden, which coincided with a higher proportion of fieldfare-redwing-songthrush samples in the central region of the country. Thus, it is possible that regional and annual variations in the prevalence of Sindbis antibodies in Swedish passerine species depend on the number of fieldfares, redwings, and songthrushes available for feeding by vector mosquitoes.

The effects of testosterone on a viral infection in greenfinches (Carduelis chloris): an experimental test of the immunocompetence-handicap hypothesis.

The immunocompetence-handicap hypothesis suggests that the honesty of quality signals could be guaranteed if testosterone (T) suppresses immune function while enhancing male ornaments. In addition, it has been proposed that the cost of enhancing ornaments should be highest for males with small ornaments. Recently, the assertion that T causes obligate immunosuppression has been questioned. In this study, we tested whether elevated T levels would increase susceptibility to a viral infection, and whether this hypothesized effect would be most pronounced in males with small ornaments. We surgically inserted T implants into 15 male greenfinches (Carduelis chloris) and control implants into a further 15 males. All birds were then infected with a naturally occurring virus (Sindbis virus, Alphavirus genus), and each bird's daily viraemia (blood virus concentration) was measured for seven days. The specific antibody response was measured for eight weeks. T-implanted males did not exhibit increased viraemia or decreased antibody response, and males with small and large ornaments did not respond differently to T implantation. We did, however, find that T implantation decreased viraemia early in the course of the infection and increased viraemia late in the infection. Thus, our results demonstrate that T may act both to increase and to decrease viraemia.

Patterns of avian seroprevalence to western equine encephalomyelitis and Saint Louis encephalitis viruses in California, USA.

Temporal and spatial changes in the enzootic activity of western equine encephalomyelitis (WEE) and St. Louis encephalitis (SLE) viruses were monitored at representative wetland study sites in the Coachella, San Joaquin, and Sacramento valleys of California from 1996 to 1998 using three methods: (1) virus isolation from pools of 50 host-seeking Culex tarsalis Coquillett females, (2) seroconversions in flocks of 10 sentinel chickens, and (3) seroprevalence in wild birds collected by mist nets and grain baited traps. Overall, 74 WEE and one SLE isolates were obtained from 222,455 Cx. tarsalis females tested in 4,988 pools. In addition, 133 and 40 seroconversions were detected in 28 chicken flocks, and 143 and 27 of 20,192 sera tested from 149 species of wild birds were positive for antibodies to WEE and SLE, respectively. WEE was active in all three valleys, whereas SLE only was detected in Coachella Valley. Seroconversions in sentinel chickens provided the most sensitive indication of enzootic activity and were correlated with seroprevalence rates in wild birds. Avian seroprevalence rates did not provide an early warning of pending enzootic activity in chickens, because positive sera from after hatching year birds collected during spring most probably were the result of infections acquired during the previous season. Few seroconversions were detected among banded recaptured birds collected during spring and early summer. Age and resident status, but not sex, were significant risk factors for wild bird infection, with the highest seroprevalence rates among after hatching year individuals of permanent resident species. Migrants (with the exception of mourning doves) and winter resident species rarely were positive. House finches, house sparrows, Gambel's quail, California quail, common ground doves, and mourning doves were most frequently positive for antibodies. The initial detection of enzootic activity each summer coincided closely with the appearance of hatching year birds of these species in our study areas, perhaps indicating their role in virus amplification. Bird species most frequently positive roosted or nested in elevated upland vegetation, sites where Cx. tarsalis host-seeking females hunt most frequently. These serosurveys provided important background information for planned host competence and chronic infection studies.

Mosquito-borne viruses in western Europe: a review.

Several mosquito-borne arboviruses belonging to the genera Alphavirus, Flavivirus, and Bunyavirus have been reported to occur in mosquitoes and to infect humans and other vertebrates in western Europe. These zoonotic viruses circulate in nature either in an Aedes-mammal, Anopheles-mammal, or Culex-bird transmission cycle. Infected humans normally do not contribute to the virus circulation. West Nile virus (Flavivirus) caused an outbreak of fever, malaise, pain in eyes and muscles, and headache and encephalitis in southern France during 1962-1965, and an outbreak of encephalitis with a high case-fatality rate in Romania during 1996. West Nile virus has been isolated from birds, horses, and mosquitoes in Portugal, France, the former Czechoslovakia, and Romania. These data, together with reports of antibodies to West Nile virus in birds, domestic mammals, and humans in several other countries, show virus activity in southern and central Europe. Sindbis virus (Alphavirus) caused outbreaks of fever, rash, and arthralgia in northern Europe during 1981-1982, 1988, and 1995. Two California group viruses (Bunyavirus), Tahyna virus and Inkoo virus, have been identified in western Europe. Tahyna virus causes fever and respiratory symptoms and sometimes also central nervous system involvement. It occurs in most countries of central and southern Europe, and is most common in central Europe. Inkoo virus has not been associated with disease in humans in western Europe although Russian studies indicated that it can cause encephalitis. Inkoo virus occurs in northern Europe, especially in the far north. Batai virus of the Bunyamwera-group (Bunyavirus) occurs in southern, central, and northern Europe, most frequently in central Europe. The antibody prevalence in humans generally is very low, indicating that the potential of this virus as a human pathogen is probably low in Europe. The Lednice virus (Bunyavirus) has been reported only from the former Czechoslovakia and Romania, and apparently is not transmitted to humans. In addition to the six mosquito-borne viruses documented in western Europe, there is serological evidence of infection with a Semliki Forest complex virus (Alphavirus) in central and southern Europe. Although mosquito-borne viruses presently are not considered to be the cause of major health problems in western Europe, the morbidity caused by Sindbis virus, and the morbidity and mortality caused by West Nile virus, merit further studies on the ecology, epidemiology, and medical importance of these viruses. The California group of viruses and a virus of the Semliki Forest complex may be the cause of unrecognized health problems in western Europe. Specific sampling of potential vectors for virus isolation, detailed characterization of virus strains, and the use of fully characterized strains for serological diagnosis will help to elucidate the present and future potential of mosquito-borne viruses as human pathogens in Europe.

Genetic relatedness of Sindbis virus strains from Europe, Middle East, and Africa.

The relatedness of 40 strains of Sindbis virus (SIN) from Europe, the Middle East, and Africa was investigated by limited sequencing within the gene encoding the E2 glycoprotein corresponding to amino acid residues 117 to 229 and encompassing one of the major neutralization epitopes. Phylogenetic analyses using distance matrix and parsimonious methods identified two major genetic clusters of western SIN strains, although the variability was less than that of the corresponding region for Venezuelan equine encephalomyelitis virus with a maximum divergence of 12.4% versus 28.5%, respectively. One cluster comprising 19 strains included the HR derivate of the Egypt SIN prototype, AR339, and strains from Israel, Saudi-Arabia, Italy, Slovak Republic, Azerbaijan, as well as three Swedish strains. Another cluster of 17 strains included the Ockelbo virus (OCK) prototype, Edsbyn 5/82, and the majority of SIN strains from northern Europe including strains from Sweden, Norway, and Karelia, as well as two strains from South Africa. A third cluster, supported by the Neighbor joining method, was made up of four strains from South Africa, Uganda, and Cameroon. Residue 212, either Ser or Thr, previously appointed important for the differences in neutralization assays between SIN and Edsbyn 5/82, respectively, correlated with the two major genetic clusters, but was a Thr for two of the three Swedish strains in the SIN prototype cluster, and a Ser in one Swedish and one Karelian strain in the OCK cluster. The finding of strains similar to prototype SIN in Middle Sweden and of strains in South Africa relating to the northern cluster of SIN strains supports the notion of the dispersal of SIN by migrating birds as previously suggested for New World alphaviruses.

Ockelbo virus (Togaviridae: Alphavirus) neutralizing antibodies in experimentally infected Swedish birds.

The ability of native Swedish birds to produce and maintain production of Ockelbo virus neutralizing (Nt) antibodies were evaluated experimentally between 6 June 1990 and 27 July 1991. After experimental infection of 57 birds of the orders Anseriformes and Passeriformes with Ockelbo virus, these birds were examined for Ockelbo virus Nt antibodies at 5 days, and at 1, 3, 6, 9, and 12 mo after inoculation. One month after inoculation, Nt antibodies were more prevalent in birds with a detectable viremia (100%, n = 22) than in non-viremic birds (65%, n = 26). The Nt antibody prevalence varied over time among taxa; detectable antibodies occurred earlier after inoculation and for a longer time in Anseriformes than in Passeriformes. By 5 days after inoculation, antibodies could be detected in 22 (71%) of 31 Anseriformes but in none of 16 Passeriformes. However, at 1 mo the antibody prevalences at 1 mo were similar: 84% among the Anseriformes and 73% among the Passeriformes; at 3 mo the prevalences were 50% in Anseriformes and 15% in Passeriformes. Forty-two percent of the Anseriformes had detectable antibodies even 12 mo after inoculation.

Antigenic comparison of Ockelbo virus isolates from Sweden and Russia with Sindbis virus isolates from Europe, Africa, and Australia: further evidence for variation among alphaviruses.

The plaque-reduction neutralization test (PRNT) was used to compare 15 isolates of Ockelbo virus from Sweden, one isolate of Ockelbo virus from Russia, the Egyptian prototype Sindbis virus, and Sindbis-like viruses from Slovakia, South Africa, Cameroon, and Australia. Strains from northern Europe (Sweden and Russia) were indistinguishable by PRNT. We observed some antigenic variation between isolates of Sindbis virus from Europe, Africa, and Australia. An Australian strain (C-377) was shown to be distinct from prototype Sindbis virus, and the Acrocephalus strain from Slovakia was shown to be identical to prototype Sindbis virus. All other strains, including Ockelbo virus isolates, were shown to be subtypes of prototype Sindbis virus.

Viremia in three orders of birds (Anseriformes, Galliformes and Passeriformes) inoculated with Ockelbo virus.

One-hundred six birds of 14 species were inoculated with approximately 10(2.7) plaque-forming units of Ockelbo virus and bled daily for 5 days to determine viremia levels. Virus was detected in birds of all 14 species tested (four Anseriformes, one Galliformes and nine Passeriformes). The onset of viremia occurred earlier and viral titers were higher in very young anseriforms and galliforms than in older birds. Adult passeriforms had Ockelbo viremias of higher titer and longer duration than did adult anseriforms. Viremia titers in adult birds of all three orders tested were sufficient to induce high transmission rates in enzootic mosquito vectors, and viremias in passeriforms could induce high transmission rates in bridging vectors as well. Passeriforms of the genera Turdus and Fringilla could serve as amplification hosts for Ockelbo virus based on the presently demonstrated viremia of high titer and long duration in these birds, and the previously demonstrated high prevalence of Ockelbo virus neutralizing antibodies in free-ranging individuals and great population size compared to birds of other taxa. Bird species of all three orders tested, however, could function as incidental hosts of the virus.

Antibodies to Ockelbo virus in three orders of birds (Anseriformes, Galliformes and Passeriformes) in Sweden.

Sera from 324 birds collected in an Ockelbo virus disease endemic area in central Sweden were examined for the presence of specific antibodies to Ockelbo virus by a plaque reduction neutralization test. Birds examined belonged to the orders Anseriformes (n = 207), Galliformes (n = 66) and Passeriformes (n = 51). Ockelbo virus neutralizing antibodies were detected in 26 (8%) of the specimens, including species from each of the three orders tested. Specific antibodies found in caged birds and in 6- to 10-week-old birds suggested local transmission. The highest antibody prevalence (27%, 14/51) was observed in the Passeriformes in which 5 of 9 species tested contained antibodies. The high antibody prevalence in passeriforms and the very large population of this group in relation to other avian groups in Sweden gives them a high potential as amplification hosts for Ockelbo virus.

Geographical and temporal distribution of Ockelbo disease in Sweden.

The incidence of Ockelbo disease and the prevalence of Ockelbo virus neutralizing antibodies were investigated in a sample of the Swedish population. The disease occurs throughout most of Sweden but with higher incidence and antibody prevalence rates in the central part of the country. It generally affects middle-aged men and women, with equal incidence between sexes, and is uncommon in people younger than 20 years of age. The disease occurs during a short period each year between the third week of July and the first week of October, with a peak during the second half of August. During the 8 years studied (1981-8), an average of 31 Ockelbo patients/year were diagnosed. The antibody prevalence rates in the oldest age groups were 20-40 times higher than the accumulated life-risk of being diagnosed and reported as an Ockelbo disease patient, which suggests that many cases are asymptomatic and/or unreported.