A new arbovirus from Aedes albopictus, an Asian mosquito established in the United States.

Ten strains of a new arbovirus belonging to the Bunyamwera group (Bunyaviridae) were recovered from field-collected Aedes albopictus mosquitoes in Potosi, Missouri. This evidence indicates that this species may serve as an arbovirus vector in the United States. The urban-suburban distribution, aggressive biting behavior, and broad viral susceptibility of Ae. albopictus may lead to the transmission of viruses of known public health importance and perhaps of viruses hitherto not transmitted to humans because of the feeding pattern of their usual vectors.

Brus Laguna virus, a Gamboa bunyavirus from Aedeomyia squamipennis collected in Honduras.

A virus isolate from Aedeomyia squamipennis collected in Honduras in 1967 was identified as a member of the Gamboa serogroup (family Bunyaviridae, genus Bunyavirus). This is the ninth Gamboa serogroup virus and the eighth shown to be a distinct serotype.

Identification of hitherto unrecognized arboviruses from Ecuador: members of serogroups B, C, Bunyamwera, Patois, and Minatitlan.

Three hundred seventy-nine virus isolates were obtained from mosquitoes collected and sentinel hamsters exposed in coastal Ecuador from 1974 to 1978. These included four alphaviruses [Venezuelan equine encephalitis 1B (1), Venezuelan equine encephalitis 1D (35), western equine encephalitis (1) and eastern equine encephalitis (4)]; two flaviviruses [St. Louis encephalitis (3) and Naranjal (6)]; 11 bunyaviruses [Maguari (243), Playas (3), Vinces (33), Turlock (2), Abras (5), Babahoyo (3), Acara (2), Guajara (3), San Juan (6), Pueblo Viejo (3), 18 unspecified Gamboa serogroup viruses, Palestina (7)]; and one vesiculovirus (vesicular stomatitis New Jersey). All but Venezuelan equine encephalitis virus were new to Ecuador, and Naranjal (serogroup B), Playas (Bunyamwera serogroup), Vinces (serogroup C), Abras and Babahoyo (Patois serogroup), San Juan and Pueblo Viejo (Gamboa serogroup) and Palestina (Minatitlan serogroup) are newly recognized viruses. These isolates have enabled us to 1) expand our knowledge of the geographic distribution of recognized viruses, 2) expand our knowledge of the members of certain serogroups and 3) establish two new serogroups (Gamboa and Minatitlan).

Reevaluation of the western equine encephalitis antigenic complex of alphaviruses (family Togaviridae) as determined by neutralization tests.

Fourteen viruses closely related to the Fleming strain of western equine encephalitis (WEE) virus were cross-tested by serum dilution-plaque reduction neutralization. The results demonstrate that strains McMillan, R-43738, AG80-646, BeAr 102091, and Y62-33 are subtypes or varieties of western equine encephalitis virus strain Fleming. Ockelbo, Kyzylagach, and Babanki are subtypes of the prototype strain (EgAr 339) of Sindbis virus. Fort Morgan and Buggy Creek viruses are closely related to each other, whereas Highlands J and Aura viruses are distinct from other members of this antigenic complex. There appear to be parallels between geographic distribution and antigenic relatedness. We hypothesize that birds, the principal vertebrate hosts for these viruses, spread the progenitor viruses north and south and from continent to continent. Viruses of the WEE complex with lesser antigenic differences may develop in discrete ecologic conditions.

Human infections with Tensaw virus in south Florida: evidence that Tensaw virus subtypes stimulate the production of antibodies reactive with closely related Bunyamwera serogroup viruses.

Maguari virus, a member of the Bunyamwera serogroup (family Bunyaviridae, genus Bunyavirus) has not been isolated north of Trinidad. Anecdotal information from other investigators has indicated the presence of antibody to Maguari virus in human residents of south Florida. We attributed such antibody to either cross-reactivity with Tensaw virus, the only Bunyamwera serogroup virus known in south Florida, or to cross-reactivity to an antigenic subtype or variant of Tensaw virus. Five strains, identified as Tensaw virus when they were isolated from mosquitoes collected in south Florida more than 20 years ago, were retrieved from storage. They were compared by serum dilution-plaque reduction neutralization tests with Bunyamwera serogroup prototypes Tensaw, Maguari, Cache Valley, and Tlacotalpan viruses. The south Florida isolates were shown to be most closely related to prototype Tensaw virus and most distantly related to prototype Maguari virus. One isolate could not be distinguished from prototype Tensaw virus, and the other 4 appeared to be subtypes of prototype Tensaw virus. More than 300 serum samples from humans in south Florida were tested for neutralizing antibody to prototypes Tensaw and Maguari viruses and to 3 of the field isolates. Thirteen had antibody to prototype Tensaw virus only, 19 to prototype Maguari virus only, and 39 to both. Antibody to all but 6 of these 71 was attributed to infection with Tensaw virus, to a subtype of Tensaw virus, or to travel or birth outside the United States. It is likely that those with antibody to Maguari virus only had been infected with yet another subtype of Tensaw virus, although another, undiscovered, Bunyamwera serogroup virus may exist in south Florida.

Arbovirus surveillance in six states during 1972.

A virus surveillance project was established and maintained during 1972 along 10 major river drainages in six states. Mosquitoes, biting flies, and blood specimens from sentinel equines were collected during 83 field trip visits to 141 arthropod collecting sites and 22 sentinel locations from April into December 1972. There were 173,074 mosquitoes tested and 303 arboviruses isolated from 11 of 41 species. From 13,388 biting flies tested, 8 arbovirus isolations were obtained in 1 of 5 species. There was no isolation of Venezuelan equine encephalitis (VEE) virus. Western equine encephalitis (WEE) virus isolates were the most numerous and were followed by Turlock, St. Louis encephalitis, Hart Park, California encephalitis, and Bunyamwera (BUN) group viruses. The first isolation of WEE from the mosquito Cullex (Mel). erraticus is reported, as is the extension of the ranges for Buttonwillow virus from California to New Mexico and Texas. Also a single isolation of the BUN group from Culicoides variipennis extends the range of this virus-vector relationship from California to Texas. New distribution records for mosquito species previously unreported for Arizona, Louisana, New Mexico, and Oklahoma are reported. The sentinel burros detected WEE serologic conversions at two sites in New Mexico and at one in Texas. The surveillance project provided state and federal officials with current information on the status of arbovirus activity, including the absence of VEE activity during 1972, and it demonstrated the existence of the potential for WEE epizootics and epidemics throughout a wide geographic area of the Western United States.

Isolation of Bhanja virus from ticks in Yugoslavia.

Two strains of Bhanja (BHA) virus, an ungrouped arbovirus, were isolated from Haemaphysalis punctata ticks collected from sheep on the island of Brac in the Yugoslavian Adriatic. Serologic testing gave evidence of the endemicity of BHA virus on the island, and antibody prevalence rates were high for both man (35.8%--ranging locally to 61.3%) and sheep (100%). No antibody was found in 82 small mammals tested. Evidence of a natural cycle involving ticks and sheep, the potential significance of this virus for man and for animals of agricultural importance, and factors related to its epidemiology are discussed.

Rio Grande--a new phlebotomus fever group virus from south Texas.

Three strains of a new Phlebotomus fever group virus were isolated from pack rats (Neotoma micropus) collected in south Texas during 1973--1974; the name Rio Grande was proposed for this virus. The virus is pH 3.0 labile, sensitive to the action of sodium deoxycholate and heat (56 degrees C) labile. The results of a serosurvey indicated that pack rats are probably the principal vertebrate host for Rio Grande virus and that year-round transmission of the virus may occur. Because no isolations of this virus were made from hematophagous insects, the vector, if any, remains undetermined.

Viruses isolated from Aedeomyia squamipennis mosquitoes collected in Panama, Ecuador, and Argentina: establishment of the Gamboa serogroup.

Twenty-four virus strains were isolated from Aedeomyia squamipennis mosquitoes collected in Ecuador. One additional strain each was isolated from this species from Panama and ARgentina. All 26 isolates were shown to be related serologically to prototype Gamboa virus, originally isolated from Ad. squamipennis mosquitoes collected in Panama. Antigenic comparisons of eight strains, including prototype Gamboa virus, indicated the existence of four distinct viruses. Neutralization tests with sera from a variety of mammalian and avian species from Argentina provided further evidence that Gamboa serogroup viruses are transmitted between Ad. squamipennis and birds.

Characterization of Fort Morgan virus, an alphavirus of the western equine encephalitis virus complex in an unusual ecosystem.

An alphavirus isolated from nestling Cliff Swallows (Petrochelidon pyrrhonota) and House Sparrows (Passer domesticus) and from cimicid bugs (Oeciacus vicarius) in eastern Colorado, for which we propose the name Fort Morgan (FM) virus, is sensitive to the action of sodium deoxycholate, unstable at pH 2.0-4.0, and demonstrates no characteristics of temperature-sensitive mutants. Unpassaged field strains are nonpathogenic, or of low pathogenicity, for suckling mice; however, plaque-purified FM virus is pathogenic for a variety of laboratory hosts. By hemagglutination-inhibition (HI), complement-fixation, and neutralization tests, cross-reactions were observed between FM virus and members of the western equine encephalitis (WEE) virus antigenic complex. Short-incubation HI tests indicated that the new isolate shared closer antigenic relationships with WEE complex virus strains from the eastern United States (Highlands J virus) than with other WEE complex viruses. On the basis of these serological findings, as well as characterization of the structural polypeptides and oligonucleotides, we suggest that FM is a distinct virus belonging to the WEE antigenic complex. A reconsideration of the taxonomy of the WEE complex and discussion of the epizoologic significance of FM virus are presented.

Distribution of Bunyamwera serogroup viruses in North America, 1956-1984.

We attempted to tabulate all Bunyamwera serogroup (family Bunyaviridae, genus Bunyavirus) isolates from North America. By summarizing information from the laboratories of the Centers for Disease Control, data generously shared by other laboratories, and the published literature, we were able to accumulate data regarding 1,372 Bunyamwera serogroup viruses. These were: Tensaw (664, including 8 from vertebrates), Cache Valley (396, including 6 from vertebrates), Main Drain (160, including 14 from vertebrates), Lokern (69, including 8 from vertebrates), Northway (13, including 5 from vertebrates), Tlacotalpan (7), Santa Rosa (2), Santa Cruz (1 from a horse), and 60 of undetermined serotype. Virus isolation rates by month of collection were correlated with collection efforts, but associations of viruses and arthropod vectors varied by location, vertebrate host, and arthropod distribution. Tensaw virus was isolated principally from Anopheles crucians mosquitoes (466/656 isolates from arthropods) in the southeastern United States; Cache Valley virus principally from An. quadrimaculatus (94), Coquillettidia perturbans (59), Culiseta inornata (45), Aedes sollicitans (30), Psorophora columbiae (23), An. punctipennis (18), and Ae. vexans and trivittatus (18 each) mosquitoes (total = 305/382 isolates from arthropods from all of the United States and Canada, except the southeastern United States); Main Drain virus from Culicoides variipennis (31), Culicoides (Selfia) sp. (65), and Psorophora (23) and Aedes (21) species mosquitoes in the western United States; Lokern virus from Culicoides species (55/61 isolates from arthropods) in the western United States. Relationships between vector and vertebrate host distributions are discussed briefly in regard to geographic distribution of the Bunyamwera serogroup viruses.

A newly recognized vesiculovirus, Calchaqui virus, and subtypes of Melao and Maguari viruses from Argentina, with serologic evidence for infections of humans and horses.

In 1983, 17 virus strains were isolated from mosquitoes collected during an outbreak of western equine encephalitis in Santa Fe Province, Argentina. Strains of western equine encephalitis, Venezuelan equine encephalitis, St. Louis encephalitis, and Antequera viruses were isolated, as were several bunyaviruses of the California and Bunyamwera serogroups and a new vesiculovirus. Complement fixation and neutralization tests were used to identify the California serogroup virus as a subtype of Melao virus, the Bunyamwera serogroup virus as a subtype of both Maguari and Playas viruses, and the vesiculovirus as a newly recognized agent for which the name Calchaqui virus is proposed. A limited serosurvey of horses and humans in Santa Fe Province and horses from the adjacent Santiago del Estero Province was performed to determine the prevalence of neutralizing antibody to the subtypes of Melao and Maguari viruses and to Calchaqui virus. The high prevalence of antibodies to these three agents indicates the need for further studies of their disease potential in horses, because they are closely related to several other viruses that are known equine pathogens.

Arbovirus investigations in Argentina, 1977-1980. III. Identification and characterization of viruses isolated, including new subtypes of western and Venezuelan equine encephalitis viruses and four new bunyaviruses (Las Maloyas, Resistencia, Barranqueras, and Antequera).

Forty viruses isolated from mosquitoes between 1977 and 1980 in Argentina have been identified and characterized. Nineteen strains of VEE virus, identical by neutralization (N) tests, were shown by hemagglutination-inhibition tests with anti-E2 glycoprotein sera to represent a new subtype VI of the VEE complex. RNA oligonucleotide fingerprints of this virus were distinct from subtype I viruses. The virus was not lethal for English short-haired guinea pigs, indicating that it is probably not equine-virulent. Three strains of a member of the WEE virus complex were shown to differ by N tests in 1 direction from prototype WEE virus. The new WEE subtype was also found to be distinct by RNA oligonucleotide mapping. Its vector relationships indicate that it is an enzootic virus, and it has not been associated with equine disease. A new member of the Anopheles A serogroup was identified, shown to be most closely related to Lukuni and Col An 57389 viruses, and given the name Las Maloyas virus. A strain of Para virus (Bunyaviridae, Bunyavirus) was identified. Six isolates, representing 3 new viruses morphologically resembling bunyaviruses are described; the names Antequera, Barranqueras, and Resistencia are proposed for these agents, which were all isolated from Culex (Melanoconion) delpontei in Chaco Province. No serologic relationships between these viruses and other bunyaviruses were found. Since they are antigenically interrelated, they form a new (Antequera) serogroup. Eight Gamboa serogroup viruses and 2 strains of St. Louis encephalitis virus were also identified.

Epizootic vesicular stomatitis in Colorado, 1982: epidemiologic studies along the northern Colorado front range.

Epidemiologic evaluations were made of farm personnel on vesicular stomatitis-affected premises along the front range of the Rocky Mountains in Colorado during the 1982 epizootic. A similar antibody prevalence was noted to that of veterinarians and research and regulatory personnel who were involved with the same epizootic. Risk of infection resulted from intimate physical contact with infected horses or cows. Incidence and infection rates in horses were 45%; rates in cows were much lower, only 5%. Some epidemiologic clues were gained by a detailed study of an equine ranch. The pasture was incriminated as the area of highest risk, where 100% infection rates were noted. Horses in open pens and barns were at lower risk. Severe clinical disease in horses resulted in higher neutralizing antibody titers than inapparent or mild infection. Maternal antibody was detected in foals up to 4 months of age, and the level of antibody in the foal was a reflection of the dam's antibody level.

Recovery of Tonate virus ("Bijou Bridge" strain), a member of the Venezuelan equine encephalomyelitis virus complex, from Cliff Swallow nest bugs (Oeciacus vicarius) and nestling birds in North America.

A second virus with distinct biological, serological, and physiochemical properties was detected as a minority viral subpopulation in specimens of Cliff Swallow nest bugs (Oeciacus vicarius) and nestling bird sera containing Fort Morgan (FM) virus. The second virus, detected by a breakthrough neutralization test employing FM antiserum, was present in 5 of 11 FM virus-positive pools of nest bugs and in 4 of 38 birds from Colorado and South Dakota. The concentration of the second virus was 10-fold to 1,000-fold lower than that of FM virus. The second virus, which was provisionally named "Bijou Bridge" (BB) virus was shown by conventional serological tests to be a member of the Venezuelan equine encephalomyelitis (VEE) complex, and by tests employing antisera to the E2 viral glycoprotein to be identical with Tonate virus, previously isolated from birds and mosquitoes only in French Guiana. Experimental infection of House Sparrows and Cliff Swallows showed that they develop brief BB viremias and antibodies. Oe. vicarius bugs were resistant to oral infection with BB virus. The epidemiological significance of recovery of Tonate virus in North American is discussed.

Identification of new Guama and Group C serogroup bunyaviruses and an ungrouped virus from Southern Brazil.

From 1975 to 1978, 36 viruses were recovered from humans, bats, birds, sentinel mice and hamsters, and from mosquitoes collected in Coastal Brazil in the state of São Paulo. Identifications of 22 of these 36 viruses have been reported. Six of the remaining 14 isolates were shown to be Guama serogroup bunyaviruses. Two of these six were strains of a newly recognized virus for which the name Cananeia virus is proposed; another is a second newly recognized Guama serogroup virus for which the name Itimirim virus is proposed; a fourth is a strain of Bertioga virus and the other two are strains of Guaratuba virus. Before these studies Guaratuba virus was considered an ungrouped bunyavirus, but cross testing by complement-fixation demonstrated that this virus, and Mirim virus as well, should be considered members of the Guama serogroup. Another six viruses were shown to be strains of a single, newly recognized Group C bunyavirus for which the name Bruconha virus is proposed. Two strains of a single virus were shown by electron microscopy to belong to the family Bunyaviridae, but serologic relationships with other members of this family of viruses were not found; the name Enseada virus is proposed for this newly recognized agent.

Bivens arm virus: a new rhabdovirus isolated from Culicoides insignis in Florida and related to Tibrogargan virus of Australia.

During field studies in 1981 on the transmission of bluetongue viruses in ruminants in Florida, a virus was isolated from Culicoides insignis collected near water buffalo (Bubalus bubalis) recently imported from Trinidad. Electron microscopy showed that this isolate, for which the name Bivens Arm virus is proposed, has rhabdovirus morphology. Serologic comparisons were made with recognized rhabdoviruses from terrestrial vertebrates and hematophagous arthropods. Indirect fluorescent antibody, complement fixation and neutralization tests indicated antigenic reactivity between Bivens Arm virus and two rhabdoviruses found only in Australia, Tibrogargan and Coastal Plains viruses. The Australian isolates cause subclinical infections in cattle and water buffalo and are believed to be transmitted by Culicoides. Initially, it was thought that Bivens Arm virus may have been introduced to Florida with the water buffalo from Trinidad, but a serologic survey of cattle serum, collected before the importation of the buffalo revealed antibody to the virus in cattle on farms located in diverse areas of Florida.

Relationship of cliff swallows, ectoparasites, and an Alphavirus in west-central Oklahoma.

Approximately 250 isolates of a newly recognized virus, related to western equine encephalitis virus (family Togaviridae, genus Alphavirus), were obtained from cimicid bugs, Oeciacus vicarius; Cliff Swallows, Hirundo pyrrhonata; and House Sparrows, Passer domesticus in a study area in west-central Oklahoma at Buggy Creek and Caddo Canyons. Antigenicity of the virus strains varied slightly from isolate to isolate. This paper summarizes the ecology of the area by describing in general the flora and fauna there.

Diagnosis of eastern equine encephalomyelitis by immunofluorescent staining of brain tissue.

Brain tissues were obtained from 5 horses with clinical encephalomyelitis during an epizootic in southwestern Michigan in August-September 1980. These tissues were tested for virus by intracerebral inoculation of suckling mice and by examination of frozen sections and impression smears by the indirect fluorescent antibody (FA) technique. Eastern equine encephalomyelitis virus was isolated and detected by FA technique in brains of 3 horses which died or were euthanatized within approximately 24 hours of onset of the disease but not from 2 horses at 2 and 3 days after onset. The latter 2 animals had serum-neutralizing antibodies at the time of death. Seven areas of the brain of 1 horse were tested. The proportion of fluorescing cells in frozen sections correlated with infectivity titers. Impression smears were negative. Viral titers ranged from 10(5.7) to 10(10.0) suckling mouse intracerebral median lethal doses/g; highest titers and most intense fluorescence were present in the thalamus and pons, emphasizing the need to obtain selective samples of central brain structures for diagnostic examination. The FA technique appears useful for the rapid diagnosis of fatal eastern equine encephalomyelitis and may be applicable in laboratories not equipped for isolation of viruses.

Serodiagnosis of western equine encephalitis virus infections: relationships of antibody titer and test to observed onset of clinical illness.

Sera from horses and human beings with clinically diagnosed western equine encephalitis (WEE) virus infections were tested for hemagglutination-inhibition (HI), complement-fixation (CF), and neutralizing (N) antibody to WEE virus. These tests confirmed infection in 43.8% (HI), 56.3% (CF), and 80.4% (N) of horses and 54.5% (HI), 59.1% (CF), and 77.3% (N) of human beings. Use of the N test as an adjunct to the HI and CF tests increased the likelihood of serologic confirmation to 91.7%. In both horses and human beings, N antibody increased steeply at the end of the 1st week after onset. The results suggested that the presence of a high HI, CF, and/or N antibody titer in a single serum obtained from horses during the acute phase of illness caused by WEE virus can be used as presumptive evidence for infection with this virus.