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PubMed1992

Expression of human salivary-gland kallikrein in insect cells by a baculovirus vector.

A cDNA fragment encoding human salivary-gland kallikrein, including the kallikrein-owned signal peptide, was inserted into a baculovirus vector adjacent to the polyhedrin promoter and expressed in transfected insect cells. Biologically active kallikrein was isolated to homogeneity from serum-free culture supernatant using a four-step protocol. The N-terminal amino acid sequence of the insect-derived kallikrein was identical to that of the natural proteinase, thus indicating the proper removal of the mammalian signal peptide.

Amino Acid Sequence↗
PubMed1994

Nematocera (Ceratopogonidae, Psychodidae, Simuliidae and Culicidae) and control methods.

The biology, veterinary importance and control of certain Nematocera are described and discussed. Culicoides spp. (family Ceratopogonidae) transmit the arboviruses of bluetongue (BT), African horse sickness (AHS), bovine ephemeral fever (BEF) and Akabane. Some other arboviruses have been isolated from these species, while fowl pox has been transmitted experimentally by Culicoides. These insects are vectors of the parasitic protozoans Leucocytozoon caulleryi and Haemoproteus nettionis, and the parasitic nematodes Onchocerca gutturosa, O. gibsoni and O. cervicalis. They also cause recurrent summer hypersensitivity in horses, ponies, donkeys, cattle and sheep. Farm animals can die as a result of mass attack by Simulium spp., which are also vectors of Leucocytozoon simondi, L. smithi and the filariae O. gutturosa, O. linealis and O. ochengi. Venezuelan equine encephalomyelitis (VEE) and Rift Valley fever (RVF) have been isolated from simuliids, and vesicular stomatitis virus New Jersey strain has been replicated in Simulium vittatum. Simuliids are well known as vectors of O. volvulus, the cause of human onchocercosis (river blindness). The family Psychodidae includes the genera Phlebotomus and Lutzomyia (subfamily Phlebotominae), vectors of Leishmania spp. in humans, dogs and other mammals. Vesicular stomatitis virus Indiana strain has been regularly isolated from phlebotomine sandflies. Mass attack by mosquitoes can also prove fatal to farm animals. Mosquitoes are vectors of the viruses of Akabane, BEF, RVF, Japanese encephalitis, VEE, western equine encephalomyelitis, eastern equine encephalomyelitis and west Nile meningoencephalitis, secondary vectors of AHS and suspected vectors of Israel turkey meningoencephalitis. The viruses of hog cholera, fowl pox and reticuloendotheliosis, the rickettsiae Eperythrozoon ovis and E. suis, and the bacterium Borrelia anserina are mechanically transmitted by mosquitoes. These insects also induce allergic dermatitis in horses. They transmit several filarial worms of both animals and humans, and are of great medical importance as vectors of major human diseases, including malaria, yellow fever, dengue fever and many more diseases caused by arboviruses.

Animals↗
PubMed2000

Densovirinae as gene transfer vehicles.

Densoviruses present an attractive opportunity to develop expression vectors for insects. They exhibit several features that are beneficial for such vectors. The genomes of densoviruses are among the smallest of animal DNA viruses, and, therefore are easier to use for cloning and transfection procedures. The fact that cloned densovirus genomes are infectious greatly simplifies their applications. It seems that both densovirus promoters have high constitutive activity and they can be further transactivated with NS. The major nonstructural protein, NS1, can retain its functions even if fused to a foreign protein. If the recombinant genome carrying a gene of interest is supplied with the missing virus functions, it can be packaged into transducing particles. The packaging capacity of the densovirus particles can accommodate a foreign gene encoding a polypeptide of up to 150 kDa. Finally, the functions missing from the recombinant densovirus genome that are necessary for the production of transducing particles can be supplied in several different ways: they can be expressed from a helper plasmid, in cells transformed with the virus genes, or from an RNA virus. The last two methods produce transducing particles which are free of wild type virus. Densovirus transducing particles can be used for the delivery and expression of a gene of interest in insect cell culture or living insects. Transducing particles expressing reporter genes can be valuable tools in the study of viral pathogenesis and perhaps in the genetic manipulation of insects.

Animals↗
PubMed1998

Evidence for transovarial transmission of tomato yellow leaf curl virus by its vector, the whitefly Bemisia tabaci.

The whitefly Bemisia tabaci is the only vector of the tomato yellow leaf curl geminivirus (TYLCV). The insect transmits the virus in a persistent-circulative manner. TYLCV DNA was detected by polymerase chain reaction and by Southern blot hybridization in progeny (eggs, first and second instars, adults) of single viruliferous whiteflies that developed on eggplant or on cotton (two TYLCV nonhost plants). Furthermore, TYLCV DNA was present in the progeny of insects that had acquired the virus through the egg. The adult progeny of the viruliferous insects and their own progeny were able to infect tomato test plants, producing typical disease symptoms. Ovaries and maturing eggs of viruliferous insects contained viral DNA, as did eggs laid by viruliferous insects maintained on an artificial diet Eggs laid by nonviruliferous whiteflies on cotton plants previously caged with viruliferous insects did not acquire viral DNA from the plant. Hence, TYLCV can be transmitted through the egg for at least two generations. In the absence of an available plant host, the whitefly may serve as a reservoir of the virus between growing seasons.

Animals↗
PubMed2004

Regional genetic differentiation of Phlebotomus sergenti in three Moroccan foci of cutaneous leishmaniasis caused by Leishmania tropica.

Phlebotomus sergenti was identified morphologically in samples from three Moroccan foci of leishmaniasis caused by Leishmania tropica in the provinces of Azilal, Essaouira and Taza. Three primary mitochondrial DNA lineages were identified, and they could be markers for regionally distributed cryptic species. Greater mitochondrial diversity in Azilal indicated that this central province could have been the origin of dispersal of P. sergenti or the zone of secondary contact. All except one of the 21 mitochondrial haplotypes showed a marked regional distribution, and this indicates that vector control would not always be followed by rapid, long-distance reinvasion. Only mitochondrial haplotype SER18 was a putative marker for long-distance dispersal, for which there is no evidence of human assistance.

Animals↗
PubMed1995

Evolutionary origin of Plasmodium and other Apicomplexa based on rRNA genes.

We have explored the evolutionary history of the Apicomplexa and two related protistan phyla, Dinozoa and Ciliophora, by comparing the nucleotide sequences of small subunit ribosomal RNA genes. We conclude that the Plasmodium lineage, to which the malarial parasites belong, diverged from other apicomplexan lineages (piroplasmids and coccidians) several hundred million years ago, perhaps even before the Cambrian. The Plasmodium radiation, which gave rise to several species parasitic to humans, occurred approximately 129 million years ago; Plasmodium parasitism of humans has independently arisen several times. The origin of apicomplexans (Plasmodium), dinoflagellates, and ciliates may be > 1 billion years old, perhaps older than the three multicellular kingdoms of animals, plants, and fungi. Digenetic parasitism independently evolved several times in the Apicomplexa.

Animals↗
PubMed1995

The transmission and geographical spread of African horse sickness and bluetongue viruses.

African horse sickness virus (AHSV) and bluetongue virus (BTV) are dsRNA viruses within the genus Orbivirus. Both are able to cause non-contagious, infectious arthropod-borne diseases in their respective vertebrate hosts. AHSV infects equines and occasionally dogs, whereas BTV replicates in ruminants. The disease caused by AHSV is usually at its most severe in horses, whereas certain breeds of sheep are particularly sensitive to BTV infection. AHSV is endemic in sub-Saharan Africa but periodically makes brief excursions beyond this area. BTV occurs much more widely and can be found in a band around the World, stretching from approximately 40 degrees N to 35 degrees S. In the wild, both viruses are transmitted between their vertebrate hosts almost entirely via the bites of arthropod vectors, although dogs can occasionally acquire AHSV by eating virus-contaminated meat and BTV may be infrequently transmitted via infected semen or transplacentally. Because of their reliance upon arthropod vectors, BTV and AHSV have a global distribution which is limited not only by the requirement for susceptible vertebrates but also by the necessity for competent arthropod vectors. The major vectors of AHSV and BTV are certain species of Culicoides biting midge, which are true biological vectors but mosquitoes and/or ticks may also be involved to a greater or lesser extent. Until recently, AHSV has apparently been unable to survive beyond its traditional endemic zones in sub-Saharan Africa for more than 2-3 years at most. This has been interpreted as being due to a number of factors, including the absence of a long-term vertebrate reservoir, a lower prevalence, shorter, seasonal incidence and decreased transmission efficiency of the local vectors and also possibly to the effect of control measures (vector abatement, vaccination). The recent outbreaks of African horse sickness (AHS) in Spain, Portugal and Morocco, which persisted for at least 5 years (1987-1991) therefore seem to have established a new pattern in AHSV survival in an epidemic zone. This extended persistence may be due to the 'all-year-round' presence in the area of adult Culicoides imicola, the major AHSV vector. This is basically an Afro-Asiatic species and its continuous presence in parts of Iberia and may be due to some recent moderation in the climate. Further northerly extensions in the range of Culicoides imicola, in response to 'climatic moderation', cannot be ruled out and could substantially increase the area of Europe 'at risk' to AHS.(ABSTRACT TRUNCATED AT 400 WORDS)

Africa South of the Sahara↗
PubMed1991

Apparent incompetence of Dermacentor variabilis (Acari: Ixodidae) and fleas (Insecta: Siphonaptera) as vectors of Borrelia burgdorferi in an Ixodes dammini endemic area of Ontario, Canada.

From April to October 1990, white-footed mice, Peromyscus leucopus (Rafinesque), were examined for ectoparasites on Long Point, Ontario, the only endemic area for Ixodes dammini Clifford, Spielman, Piesman & Corwin and Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner known in Canada. Larval and nymphal I. dammini and Dermacentor variabilis (Packard), and adult fleas Orchopeas leucopus (Baker), Epitedia wenmanni (Rothschild), and Ctenophthalamus pseudagrytes Baker were common on trapped mice. Questing ticks were collected by dragging, near the sites of mouse trapping, from April to November 1990. Indirect immunofluorescent assay established that 58.3% of adult, 17.3% of nymphal, and 0.15% of larval I. dammini questing at Long Point were infected with B. burgdorferi, indicating that infected mammalian reservoir hosts are common. None of 593 adult, 2 nymphal, and 4 larval D. variabilis collected while questing were infected; and only 1 of 322 fleas (O. leucopus) removed from white-footed mice was infected. The fact that no unfed adult D. variabilis and only one flea were infected, in a situation where the probability of exposure of hematophagous ectoparasites is moderately high, suggests that this species of tick and the fleas examined are poor vectors for the Lyme disease spirochete.

Animals↗
PubMed2024

Characterization of West Nile virus Koutango lineage from phlebotomine sandflies in Kenya.

The West Nile virus (WNV), primarily transmitted by mosquitoes, is one of the most widespread flaviviruses globally, with past outbreaks occurring in the USA and Europe. Recent studies in parts of Africa, including Kenya, have identified the West Nile virus Koutango lineage (WN-KOUTV) among phlebotomine sandfly populations, however, our understanding of this virus remains limited. This study aimed to characterize WN-KOUTV from phlebotomine sandflies. Sandflies were sampled between 12th -16th March 2021 and 16th -20th March 2023 from six villages each in Baringo and Isiolo Counties, using CDC light traps. Female sandflies were taxonomically identified and pooled based on genus and site of collection. Virus isolation was performed in Vero cells. Viral genomes were determined using next-generation sequencing. Phylogenetic and molecular clock analyses were done to decipher the virus's evolutionary relationships. Comparative analyses of amino acid sequences were performed to determine variations. Protein modeling in Pymol was conducted to elucidate variations in key protein regions. Evolutionary pressure analysis investigated the selection pressures on the virus. In vitro experiments were done to investigate the virus growth kinetics in mammalian Vero E6 and mosquito C6/36 cells. We report the isolation of WN-KOUTV from Salabani in Baringo and Aremet in Isiolo, Kenya. The isolated WN-KOUTVs clustered with previously identified WN-KOUTV strains. Comparative analysis revealed a unique amino acid at NS5 653. The WN-KOUTV lineage as a whole is under purifying selective pressure, with diversifying pressure acting at site NS3 267. The current WN-KOUTV replicated in Vero E6 and C6/36 cells comparable to West Nile virus Lineage 1a, isolated from mosquitoes. Subsequent isolations of WN-KOUTV in phlebotomine sandflies suggest potential vectors, however, vector competence studies would confirm this. Replication in mammalian and insect cell lines suggests there may exist a vector/host relationship. We speculate the close genetic relationship of WN-KOUTV strains from East and West Africa may potentially be enabled by bird migratory routes between the two regions. If proven, this could point to a potential future pandemic pathway for this virus.

Animals↗
PubMed2003

Japanese encephalitis vaccines: moving away from the mouse brain.

Japanese encephalitis (JE) is a severe disease that is widespread throughout Asia and is spreading beyond its traditional boundaries. Three vaccines are currently in use against JE but only one is available internationally, a mouse-brain-derived inactivated vaccine first used in the 1930s. Although this vaccine has been effective in reducing the incidence of JE, it is relatively expensive and has been linked to severe allergic and neurological reactions. Cell-culture-derived inactivated and attenuated vaccines have been developed but are only used in the People's Republic of China. Other vaccines currently in various stages of development are DNA vaccines, a chimeric yellow fever-JE viral vaccine, virus-like particle vaccines and poxvirus-based vaccines. Poxvirus-based vaccines and the chimeric yellow fever-JE vaccine have been tested in Phase I clinical trials. These new vaccines have the potential to significantly reduce the impact of JE in Asia, particularly if used in an oral vaccine delivery strategy.

Administration, Oral↗
PubMed2003

[Potential of Bacillus thuringiensis israelensis Berliner for controlling Aedes aegypti].

The importance of the entomopathogenic bacterium Bacillus thuringiensis israelensis in the control of Aedes aegypti is presented. The use and potential of B. thuringiensis israelensis against the mosquito vector of dengue fever is described. Other aspects such as insect's resistance development against chemicals and advantages and constraints of using microbial control are discussed. Emphasis is given to the importance of the use of this bacterium in Brazil, which could contribute significantly to solving the mosquito problem without affecting the environment, humans and others invertebrate organisms in critical regions.

Aedes↗