Mechanisms of arthropod transmission of plant and animal viruses.
A majority of the plant-infecting viruses and many of the animal-infecting viruses are dependent upon arthropod vectors for transmission between hosts and/or as alternative hosts. The viruses have evolved specific associations with their vectors, and we are beginning to understand the underlying mechanisms that regulate the virus transmission process. A majority of plant viruses are carried on the cuticle lining of a vector's mouthparts or foregut. This initially appeared to be simple mechanical contamination, but it is now known to be a biologically complex interaction between specific virus proteins and as yet unidentified vector cuticle-associated compounds. Numerous other plant viruses and the majority of animal viruses are carried within the body of the vector. These viruses have evolved specific mechanisms to enable them to be transported through multiple tissues and to evade vector defenses. In response, vector species have evolved so that not all individuals within a species are susceptible to virus infection or can serve as a competent vector. Not only are the virus components of the transmission process being identified, but also the genetic and physiological components of the vectors which determine their ability to be used successfully by the virus are being elucidated. The mechanisms of arthropod-virus associations are many and complex, but common themes are beginning to emerge which may allow the development of novel strategies to ultimately control epidemics caused by arthropod-borne viruses. (+info)
Genetic and fitness changes accompanying adaptation of an arbovirus to vertebrate and invertebrate cells.
The alternating host cycle and persistent vector infection may constrain the evolution of arboviruses. To test this hypothesis, eastern equine encephalitis virus was passaged in BHK or mosquito cells, as well as in alternating (both) host cell passages. High and low multiplicities were used to examine the effect of defective interfering particles. Clonal BHK and persistent mosquito cell infections were also evaluated. Fitness was measured with one-step growth curves and competition assays, and mutations were evaluated by nucleotide sequencing and RNA fingerprinting. All passages and assays were done at 32 degrees C to eliminate temperature as a selection factor. Viruses passaged in either cell type alone exhibited fitness declines in the bypassed cells, while high-multiplicity and clonal passages caused fitness declines in both types of cells. Bypassed cell fitness losses were mosquito and vertebrate specific and were not restricted to individual cell lines. Fitness increases occurred in the cell line used for single-host-adaptation passages and in both cells for alternately passaged viruses. Surprisingly, single-host-cell passage increased fitness in that cell type no more than alternating passages. However, single-host-cell adaptation resulted in more mutations than alternating cell passages. Mosquito cell adaptation invariably resulted in replacement of the stop codon in nsP3 with arginine or cysteine. In one case, BHK cell adaptation resulted in a 238-nucleotide deletion in the 3' untranslated region. Many nonsynonymous substitutions were shared among more than one BHK or mosquito cell passage series, suggesting positive Darwinian selection. Our results suggest that alternating host transmission cycles constrain the evolutionary rates of arboviruses but not their fitness for either host alone. (+info)
Establishment and characterization of a new continuous cell line from Lutzomyia longipalpis (Diptera: psychodidae) and its susceptibility to infections with arboviruses and Leishmania chagasi.
Embryonic tissue explants of the sand fly Lutzomyia longipalpis (Lutz & Neiva 1912) the main vector of Leishmania chagasi (Cunha and Chagas), were used to obtain a continuous cell line (Lulo). The tissues were seeded in MM/VP12 medium and these were incubated at 28 masculineC. The first subculture was obtained 45 days after explanting and 96 passages have been made to date. Lulo is composed of epithelioid cells, showed a 0.04 generations/hour exponential growth rate and population doubling time at 24.7 h. The cell line isoenzymatic profiles were determined by using PGI, PGM, MPI and 6-PGDH systems, coinciding with patterns obtained from the same species and colony's pupae and adults. The species karyotype characteristics were recognized (2n = 8), in which pair 1 is subtelocentric and pairs 2, 3 and 4 are metacentric. Lulo was free from bacterial, fungal, mycoplasmic and viral infection. Susceptibility to five arbovirus was determined, the same as Lulo interaction with Leishmania promastigotes. (+info)
Complete sequence determination and genetic analysis of Banna virus and Kadipiro virus: proposal for assignment to a new genus (Seadornavirus) within the family Reoviridae.
Arboviruses with genomes composed of 12 segments of double-stranded (ds) RNA have previously been classified as members or probable members of the genus Coltivirus within the family REOVIRIDAE: A number of these viruses have been isolated in North America and Europe and are serologically and genetically related to Colorado tick fever virus, the Coltivirus type species. These isolates constitute subgroup A of the coltiviruses. The complete genome sequences are now presented of two Asian arboviruses, Kadipiro virus (KDV) and Banna virus (BAV), which are currently classified as subgroup B coltiviruses. Analysis of the viral protein sequences shows that all of the BAV genome segments have cognate genes in KDV. The functions of several of these proteins were also indicated by this analysis. Proteins with dsRNA-binding domains or with significant similarities to polymerases, methyltransferases, NTPases or protein kinases were identified. Comparisons of amino acid sequences of the conserved polymerase protein have shown that BAV and KDV are only very distantly related to the subgroup A coltiviruses. These data demonstrate a requirement for the subgroup B viruses to be reassigned to a separate new genus, for which the name Seadornavirus is proposed. (+info)
Kaeng Khoi virus from naturally infected bedbugs (cimicidae) and immature free-tailed bats.
Kaeng Khoi virus was recovered from bedbugs (Stricticimex parvus and Cimex insuetus) and from suckling wrinkle-lipped bats (Tadarida plicata) collected in central Thailand. The data implicate bedbugs as possible vectors of this virus. (+info)
Snail control in urban sites in Brazil with slow-release hexabutyldistannoxane and pentachlorophenol.
Slow release formulations of hexabutyldistannoxane (TBTO) and pentachlorophenol (PCP) were tested for the control of Biomphalaria tenagophila in 52 urban sites in Rio de Janeiro. TBTO acted faster and lasted longer than PCP and at 15 g/m(2) it eliminated snails from 76% of the treated sites for 1 year. Water pollution and rate of flow had no significant influence on the molluscicidal properties of either compound, but alkalinity lowered the activity of TBTO. Failure to control snail populations was due mainly to human interference and to the non-treatment of adjacent breeding sites that were temporarily dry and therefore overlooked. (+info)
Assay of togavirus haemagglutination-inhibition antibodies by the micro method: loss of information and its rectification.
Haemagglutination inhibition (HI) activity was assayed by the micro and macro methods in immune sera prepared against four togaviruses. HI titres were always 4 to 8 times lower by the micro method, and coincided with 4 to 8 times lower haemagglutinin titres in micro method assays. Because of this phenomenon, positive sera with HI macro method titres lower than 1: 80 will be false negative for HI by the micro method when tests begin at a 1: 10 serum dilution. This was confirmed for a number of human sera tested for West Nile virus HI activity. As a consequence of the difference between titres given by the two assay methods, results of seroepidemiological studies may be distorted. Use of a system that combines both the macro and micro methods could rectify this distortion. (+info)
Inhibition of group B arbovirus antigen production and replication in cells enucleated with cytochalasin B.
A comparative study of the growth of Sindbis (SIN) virus, a group A arbovirus (togavirus), and Japanese encephalitis (JE) virus, a representative group B arbovirus (togavirus), was conducted in enucleate and nucleate cells. Immunofluorescent tests and yield measurements demonstrated that chicken embryo cells which had been enucleated and subsequently infected with SIN virus produced virus-specific antigens and infectious virus. By contrast, JE failed to replicate or produce virus-specific antigen in cells which had been enucleated before or even 2 h post infection. Studies of the effect of enulceation at various times after infection demonstrated that a nucleus must be present at least 2 and possibly as long as 4 h after infection to produce either JE-specific antigen or infectious JE virus. These studies demonstrate that the replication of SIN, a group A arbovirus (togavirus), which has no nuclear requirement, contrasts sharply with that of a group B arbovirus (togavirus), JE, which may have an initial dependence on a nucleus-associated process. (+info)