Phlebotomus fever in Egypt. Isolation of phlebotomus fever viruses from Phlebotomus papatasi.
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The antigenic relationships of 21 known or presumed Phlebotomus fever group serotypes and of 2 ungrouped, solvent sensitive, sandfly-associated arboviruses (Pacui and Charleville) were studied by complement fixation, plaque neutralization, and hemagglutination-inhibition methods. Results of complement fixation and neutralization tests were specific, allowing clear separation of the various serotypes, while those of the hemagglutination-inhibition test showed broader crossing and lack of specificity. Pacui virus was shown to be a member of the Phlebotomus fever serogroup. Six new Phlebotomus fever group serotypes are also described, increasing the known members of the group to 22. The implications of these and other recent data about the epidemiology of sandfly fever are discussed.
Human and animal sera from an endemic area of sandfly fever in Iran were tested by plaque reduction neutralization method against five different Phlebotomus fever virus serotypes (Naples, Sicilian, Karimabad, Salehabad, and I-47). The overall prevalence of Naples, Sicilian, and Karimabad virus antibodies among the human population was 17%, 25%, and 66%, respectively. All sera were negative against Salehabad and I-47 viruses. Age-specific antibody rates suggested that Sicilian and Karimabad viruses were endemic in the study area but that Naples virus activity was sporadic. These observations were confirmed by isolations of Sicilian and Karimabad viruses from sandflies collected in the study area. Among the animal sera tested, evidence of Phlebotomus fever virus infection was detected only in gerbils. Of 38 Rhombomys opimus tested, 34% had neutralizing antibodies against Sicilian virus and 32% against Karimabad. These results indicate that gerbils are infected with these two viruses and possibly might serve as reservoirs or amplifying hosts. The serologic studies also suggest that the ecology of Sicilian and Karimabad viruses involves chiefly sandflies, gerbils, and man, an epidemiologic pattern previously demonstrated for cutaneous leishmaniasis in the same region of Iran.
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Twenty volunteers were inoculated with various doses of human serum containing Phlebotomus fever virus (Sicilian type) to determine their clinical and serologic responses as well as the human infectious dose50 of the virus. All infected subjects developed fever which varied in duration from 6 to 74 hours. The most common symptoms during sandfly fever were headache, anorexia, myalgia, photophobia, low back and retro-orbital pain. Infected individuals developed a marked leukopenia characterized by an initial lymphopenia followed by protracted neutropenia. Little complement fixing antibody was detected in convalescent sera but most subjects developed significant rises in hemagglutination inhibiting antibodies. All infected subjects developed specific neutralizing antibodies with titers ranging from 1:40 to 1:2,560. Of the three serologic tests performed, the plaque reduction neutralization method appears to be the most sensitive test for detecting antibodies to Phlebotomus fever viruses.
Eight new members of the phlebotomus fever arbovirus serogroup (family Bunyaviridae; genus Phlebovirus) from the Amazon region of Brazil are described. One serotype was recovered from a febrile patient, three from small wild animals and four from sand flies. A small serum survey carried out with the human isolate, Alenquer virus, suggests that it rarely infects man. Complement-fixation and plaque reduction neutralization tests were done, comparing the eight new viruses with other members of the phlebotomus fever serogroup. A close antigenic relationship was demonstrated between one of the new agents (Belterra) and Rift Valley fever virus. This finding is of considerable interest and deserves further investigation. Addition of these eight new viruses to the genus Phlebovirus brings to 14 the number of serotypes known to occur in the Amazon region and to 36 the total number reported worldwide. More detailed clinical and epidemiological studies should be conducted in Amazonia in order to define the public health impact caused by phleboviruses.
Toscana (TOS) virus, a new member of the Phlebotomus fever serogroup of arboviruses, was isolated from sandflies collected in Italy. It was shown to be closely related to Sandfly fever Naples (SFN) virus. The antigenic relationships between the two viruses were analysed in the complement fixation (CF), plaque reduction neutralization (PRN) and indirect fluorescent antibody (IFA) tests. TOS virus multiplied to high titres with a cytopathic effect (CPE) in several vertebrate cell cultures (e.g. Vero, BHK-21, etc.), whereas it failed to replicate in mosquito cell cultures. Plaque formation by TOS virus in Vero cells was not affected by incorporating dimethyl sulphoxide in the overlay, which is known to enhance plaque formation by SFN virus. Some biological characteristics in cell cultures and in mice of a mouse-adapted and a cell-adapted TOS virus strain were studied. After 4-6 passages in the two different systems, changes in plaque size, in the amount of autointerfering activity and of virulence for mice were observed.
Six isolates of a new phlebotomus fever serogroup virus, designated Arboledas virus, were obtained from sand flies (Lutzomyia spp.) collected in northeastern Colombia. One of the isolates was made from a pool of male sand flies. By immunofluorescence, Arboledas virus is related to Caimito and Pacui viruses; by neutralization test, it is distinct. Arboledas virus neutralizing antibodies were found in the sera of opossums (Didelphis marsupialis) and humans living in the study area. D. marsupialis inoculated with the virus developed a viremia of four days' duration, and sand flies (Lutzomyia gomezi) feeding on a viremic opossum were readily infected. Transovarial transmission of Arboledas virus was also demonstrated in experimentally infected Lu. gomezi. Results of the above laboratory studies suggest that Arboledas virus is maintained in nature by two mechanisms: vertical (transovarial) transmission in the insect vector, and an alternating marsupial-sand fly cycle. The implications of this complex maintenance cycle for other phleboviruses are discussed.
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.
Analyses of the virion polypeptides and genomes of several Phlebotomus fever group viruses, Karimabad, Punta Toro, Chagres, and the sandfly fever Sicilian serotype viruses, have established that they are biochemically similar to the accepted members of the Bunyaviridae family. Like snowshoe hare virus (a member of the California serogroup of the Bunyavirus genus of the Bunyaviridae family), Karimabad, Punta Toro, Chagres, and the sandfly fever Sicilian serotype viruses all have three viral RNA species, designated large (L), medium (M), and small (S). Oligonucleotide fingerprint analyses of Karimabad and Punta Toro virus RNA species indicated that their L, M, and S RNA species are unique. By polyacrylamide gel electrophoresis it was determined for Karimabad virus that the apparent molecular weights of its L, M, and S RNA species are 2.6 X 10(6), 2.2 X 10(6), and 0.8 X 10(6), respectively. For Punta Toro virus, the apparent molecular weights of its L, M, and S RNA species are 2.8 X 10(6), 1.8 X 10(6), and 0.75 X 10(6), respectively. The major internal nucleocapsid (N) protein of Karimabad virus was found to have a molecular weight of 21 X 10(3). A similar polypeptide size class was identified in preparations of sandfly fever Sicilian serotype, Chagres, and Punta Toro viruses. The Karimabad virus glycoproteins formed the external surface projections on virus particles and could be removed from virus preparations by protease treatment. The glycoproteins in an unreduced sample could be resolved into two size classes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. They had apparent molecular weights of 62 X 10(3) and 50 X 10(3) in continuous polyacrylamide gels. When Karimabad virus preparations were reduced with 1% beta-mercaptoethanol, prior to resolution by continuous polyacrylamide gel electrophoresis, all the viral glycoprotein was recovered in a single size class, having an apparent molecular weight of 62 X 10(3). Two or three major virion polypeptides have been identified in preparations of Punta Toro, Chagres, and sandfly fever Sicilian serotype viruses.
Dimethyl sulfoxide (DMSO)incorporated into an agar overlay containing DEAE-dextran enhanced plaque formation in Vero cells by Naples sandfly fever virus passaged in mouse brain or Vero cell cultures. No plaques were visible when DMSO was used without the DEAE-dextran, some plaques were rarely visible (less than 0.5mm) when DEAE-dextran was used without the DMSO, and up to 10-fold more plaques were clearly visible (0.5-1.5 mm) when both chemicals were used. The combined enhancing effect of DMSO and DEAE-dextran was also observed with mouse brain passaged, but not Vero passaged Sicilian sandfly fever virus. Other Phlebotomus group viruses produced a bit plaques (3-5 mm) and did not require DMSO for plaque formation, although an increase in plaque clarity was obtained with DMSO for some of them. Plaque reduction neutralization tests were assayed successfully under agar containing DMSO. The alphavirus Sindbis produced slightly larger plaques under agar containing DMSO, but there was no effect on clarity or size of plaques produced by the flavivirus dengue-2.
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