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PubMed2000

Application of PCR assays to determine the genotype of Babesia bovis parasites isolated from cattle with clinical babesiosis soon after vaccination against tick fever.

OBJECTIVE: To demonstrate the value of PCR assays to determine the genotypes of Babesia bovis in cattle with clinical signs of babesiosis within 3 weeks after vaccination against tick fever. DESIGN: Samples from 5 cases of babesiosis in cattle soon after vaccination against tick fever were analysed in two PCR assays. PROCEDURE: Parasite DNA was purified from blood taken from cattle with signs of babesiosis within 3 weeks of vaccination against tick fever. DNA was also prepared from the tissues of animals that died of babesiosis. Two PCR assays that amplify repeat sequences of DNA within the B bovis genes, Bv80 and BvVA1, were used to differentiate the genotypes of field isolates and vaccine strains of B bovis. RESULTS: One of the five cases of babesiosis was found to be caused by a vaccine strain, but PCR analyses showed that the predominant isolate in the other four cases was not the vaccine strain. CONCLUSIONS: PCR assays on the DNA of B bovis obtained from the blood or tissues of cattle clinically affected with tick fever within 3 weeks after vaccination are useful to distinguish between vaccine strains and field isolates as the source of infection.

Animals↗
PubMed1985

Diagnosis of Colorado tick fever virus infection by enzyme immunoassays for immunoglobulin M and G antibodies.

An immunoglobulin M (IgM) capture enzyme immunoassay technique was adapted for the detection of antibody to Colorado tick fever virus in sera from 84 individuals for whom diagnosis had been confirmed by virus isolation or neutralization test. Titers were compared with those for IgG and neutralizing antibodies in these Colorado tick fever cases. IgM antibody titers were higher than neutralizing antibody titers, but neither appeared until 1 to 2 weeks after the onset of illness. Neutralizing antibodies were detected earlier than IgM antibodies, and both were detected with greater frequency than IgG antibodies. Late-convalescent-phase sera contained both neutralizing and IgG antibodies, but IgM was all but undetectable by 2 months after onset. Although the neutralization test may remain the serological test of choice, the enzyme immunoassay for IgM antibody offers a simple and more rapid method of serodiagnosis; the enzyme immunoassay is, however, less sensitive than the neutralization test. Furthermore, because there was a sharp decline in IgM antibody after 45 days, the presence of IgM antibody in a single serum sample provides a basis for the presumptive serodiagnosis of recent Colorado tick fever virus infection.

Adolescent↗
PubMed1964

PLAQUE ASSAY PROCEDURE FOR COLORADO TICK FEVER VIRUS.

Deig, E. Frank (University of California, Berkeley), and H. M. S. Watkins. Plaque assay procedure for Colorado tick fever virus. J. Bacteriol. 88:42-47. 1964.-A reproducible plaque assay procedure is described for the quantitation of Colorado tick fever virus in a cell line established from embryonic hamster tissue. Under the best conditions, plaques approximately 4 mm in diameter were formed after incubation at 37 C of 4 to 6 days. Several environmental variables in the procedure were studied. Efficiency was increased markedly by combining the virus during adsorption with serum proteins, and by carrying out this step at 25 C rather than at 37 C. The overlay medium used contained metabolites which promoted cell viability for periods greater than 1 week, and allowed plaques to develop. Plaque formation was relatively insensitive to a variation in pH between 7.1 and 8.1 (with optimal concentrations of bicarbonate). However, plaque development was inhibited with medium containing greater than 0.22% bicarbonate (at optimal pH), or when the initial pH was less than 7.0.

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PubMed1997

Detection of Colorado tick fever virus by using reverse transcriptase PCR and application of the technique in laboratory diagnosis.

Colorado tick fever (CTF) virus elicits an acute illness in humans, producing nonspecific flu-like symptoms and a biphasic fever in approximately 50% of patients. The disease is transmitted by the adult Rocky Mountain wood tick (Dermacentor andersoni), and therefore incidence is limited by the habitat and life cycle of that vector. The early symptoms of infection are difficult to distinguish from those of several other agents, especially Rickettsia rickettsii. Serologic testing is usually unable to provide evidence of CTF viral infection during the acute phase because of the late appearance of the various antibodies. Here we report the development and clinical application of a test to diagnose this disease during the acute stages. Oligonucleotide primers to the S2 segment of CTF (Florio) virus were made, and these were used in the amplification of a 528-bp fragment of DNA, transcribed from the double-stranded CTF virus RNA template by reverse transcriptase PCR. RNAs processed from 16 CTF virus isolates yielded similar results when analyzed on agarose gels. These were distinguishable from their antigenic relatives Eyach, S6-14-03, and T5-2092 and from other coltiviruses and an orbivirus but not from the antigenically distinct CTF virus-related isolate 720896. A mouse model demonstrated the utility of this method with whole-blood specimens, and CTF virus was successfully detected in human sera from the initial day of the onset of symptoms to 8 days later. The reverse transcriptase PCR method is a promising tool for the early diagnosis of CTF viral infection, or for ruling out CTF virus as the etiologic agent, in order to facilitate appropriate medical support.

Adult↗
PubMed2004

Termination and read-through proteins encoded by genome segment 9 of Colorado tick fever virus.

Genome segment 9 (Seg-9) of Colorado tick fever virus (CTFV) is 1884 bp long and contains a large open reading frame (ORF; 1845 nt in length overall), although a single in-frame stop codon (at nt 1052-1054) reduces the ORF coding capacity by approximately 40 %. However, analyses of highly conserved RNA sequences in the vicinity of the stop codon indicate that it belongs to a class of 'leaky terminators'. The third nucleotide positions in codons situated both before and after the stop codon, shows the highest variability, suggesting that both regions are translated during virus replication. This also suggests that the stop signal is functionally leaky, allowing read-through translation to occur. Indeed, both the truncated 'termination' protein and the full-length 'read-through' protein (VP9 and VP9', respectively) were detected in CTFV-infected cells, in cells transfected with a plasmid expressing only Seg-9 protein products, and in the in vitro translation products from undenatured Seg-9 ssRNA. The ratios of full-length and truncated proteins generated suggest that read-through may be down-regulated by other viral proteins. Western blot analysis of infected cells and purified CTFV showed that VP9 is a structural component of the virion, while VP9' is a non-structural protein.

Animals↗
PubMed1993

Replication of Colorado tick fever virus within human hematopoietic progenitor cells.

Significant neutropenia, as well as thrombocytopenia and a mild anemia, occurs in patients infected with Colorado tick fever virus. In this study, human bone marrow CD34+ cells and KG-1a cells, a human hematopoietic progenitor cell line, were infected in vitro with Colorado tick fever virus. The time course and morphological appearance of viral replication in human progenitor cells were similar to those seen in erythroblasts and in HEL cells and suggest one possible mechanism for the clinical hematologic findings.

Animals↗
PubMed1981

The ecology of Colorado tick fever in Rocky Mountain National Park in 1974. II. Infection in small mammals.

Field studies of Colorado tick fever (CTF) in small mammals in Rocky Mountain National Park (RMNP) in 1974 established that Eutamias minimus and Spermophilus lateralis were the most important hosts for CTF virus and were the source of virus for immature stages of the tick vector, Dermacentor andersoni. Other species (Peromyscus maniculatus, Spermophilus richardsonii, Eutamias umbrinus) are secondary hosts. The intensity of viral activity in rodents varied greatly from locality to locality. Highest rodent infection rates were found to occur in the Moraine Park area of RMNP. Lowest infection rates occurred above 3,290 meters in altitude at Rainbow Curve and on the tundra. The prevalence of infection in rodents was constant from April--July (5--6% of animals captured were viremic) and then declined to 1.7--2.5% in August and September coincident with a decline in nymphal tick ectoparasitism. Many animals were captured which were simultaneously viremic and antibody-positive. Under field conditions, neutralizing antibody seroconversion does not always occur.

Animals↗
PubMed1978

The development of Colorado tick fever virus within cells of the haemopoietic system.

Electron microscopic examination of haemopoietic liver tissue from mice infected in utero or when newborn showed inclusions of Colorado tick fever virus within erythroblasts, reticulocytes and erythrocytes. Inclusions were also seen within erythroblastoid cells undergoing mitosis. Other evidence of virus replication within erythropoietic cells was the presence of intracytoplasmic and intranuclear fibres, which have been shown to be associated with Colorado tick fever virus replication. The findings reported here support the hypothesis that virus replication within infected erythropoietic cells occurs concurrently with differentiation of the infected cell, resulting in the presence of virions within erythrocytes.

Animals↗
PubMed1976

Eyach--an arthropod-borne virus related to Colorado tick fever virus in the Federal Republic of Germany.

From Ixodes ricinus ticks collected in a tick-borne encephalitis focus in Baden-Württemberg, an agent pathogenic exclusively for suckling mice was isolated. Obviously a virus, it passed membrane filters with a pore width of 200 nm. It was resistant to ether and sodium deoxycholate, but not to chloroform. In the complement fixation test it showed a close, and in the neutralisation test a more one-sided relationship to Colorado tick fever virus.

Animals↗
PubMed1982

Survey for evidence of Colorado tick fever virus outside of the known endemic area in California.

A virus very similar or identical to Colorado tick fever (CTF) virus was recovered from the blood clot of one of 104 black-tailed jack rabbits (Lepus californicus) examined during a survey for various zoonotic agents in mammals and ticks from the University of California, Hopland Field Station, Mendocino County, California, 1974--79. This is the first reported isolation of a CTF-like virus from L. californicus, and only the second time such a virus has been found in northwestern California. Mendocino County is located far outside the known distributional ranges of the most common mammalian hosts of CTF virus and of Dermacentor andersoni, the only proven tick vector for man. The viral isolate is very similar to a CTF-like virus previously recovered from the blood and spleen of a western gray squirrel (Sciurus griseus) from San Luis Obispo County, an area also outside of the previously-known CTF area. Virus was not isolated from 14 additional species of mammals (354 specimens) or from eight species of ticks (4,487 individuals), but CTF-neutralizing antibodies were detected in 28 of 771 (3.6%) sera from seven of 15 mammalian species including significant titers (greater than or equal to 1:8) in two species and one subspecies not previously reported as natural hosts, i.e., brush mouse (Peromyscus boylii), pinyon mouse (P. truei), and Columbian black-tailed deer (Odocoileus hemionus columbianus). CTF indirect immunofluorescent antibodies also were detected in 26 of 129 (20.2%) sera belonging to four of five mammalian species tested. Neutralizing antibodies were found in sera of deer from other localities in Mendocino County, from a deer mouse from Napa County, and from a brush rabbit from Monterey County as well. These findings suggest that a virus identical or similar to CTF virus is widespread in northwestern-westcentral California, and that surveillance for human cases of CTF or a similar disease should be extended to cover this region.

Animals↗
PubMed1964

COLORADO TICK FEVER VIRUS IN CELL CULTURE. I. CELL-TYPE SUSCEPTIBILITY AND INTERACTION WITH L CELLS.

Trent, Dennis W. (University of Oklahoma School of Medicine, Oklahoma City) and L. Vernon Scott. Colorado tick fever virus in cell culture. I. Cell-type susceptibility and interaction with L cells. J. Bacteriol. 88:702-708. 1964.-Colorado tick fever (CTF) virus was serially propagated in monolayer cultures of L and FL cells. Early passages of virus in FL cells yielded viral titers 10(4)-fold greater than did the corresponding L-cell passages. During L-cell passage number 4, there was a 10(3)-fold increase in the amount of infectious virus produced as compared with virus cultured earlier in this cell line. Viruses from L-cell passages 8 and 12 were identified with specific immune serum to be CTF viruses which were antigenically similar, if not identical, to the mouse-adapted virus. Parallel titrations of mouse-, L cell-, and FL cell-adapted viruses were performed in mice and replicate monolayers of L, FL, HeLa, KB, chick embryo, and cotton rat kidney cells. Cytopathic effects and viral replication were noted in all cultures except HeLa and cotton rat kidney. Cultures of L, FL, and chick embryo cells were as sensitive to infection as were suckling or weanling mice. KB cells were the least susceptible of those cell types examined. In L-cell cultures, 90% of the input virus was adsorbed to the cells during the first 30 min of incubation. The latent period lasted 10 to 12 hr, and was followed by rapid viral synthesis for the next 10 to 24 hr, depending upon the multiplicity of infection. Curves describing exponential increase in cell-associated and cell-released virus were separated by 4 hr. When the maximal total virus titers were reached, 80 to 90% of the virus was released from the cell.

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PubMed1993

Ecology of porcupines (Erethizon dorsatum) and Colorado tick fever virus in Rocky Mountain National Park, 1975-1977.

The involvement of porcupines, Erethizon dorsatum (L.), in the ecology of Colorado tick fever (CTF) virus in Rocky Mountain National Park was investigated from 1975 to 1977. Porcupine dens and feeding activity were found mostly on rocky knolls or on south-facing slopes within open stands of the montane coniferous forest, and 20 adult porcupines were trapped or captured by hand at those locations. An average of 24.6 +/- 5.4 adult Dermacentor andersoni Stiles ticks were found per animal (annual range, 17.5-31.4 ticks). The minimum CTF virus infection rate of the ticks removed from porcupines varied from 129 to 257, whereas for questing adult D. andersoni ticks from the same geographic area was 205 in 1976. No virus was isolated from the 20 animals, but 85% had neutralizing antibody against CTF virus. Porcupines utilize the same habitats described for the CTF ecosystem in Rocky Mountain National Park and appear to be an important host for adult D. andersoni.

Animals↗
PubMed1975

Teratogenic effects of Colorado tick fever virus in mice.

The teratogenic effects in mice of Colorado tick fever (CTF) virus, a human pathogen, were demonstrated. Various routes of inoculation, gestational periods, and dosages of virus were studied. The incidence of stillbirth and neonatal death increased significantly after inoculation of pregnant mice with CTF virus. CTF virus crossed the placenta and replicated to high titers in the placental and fetal tissues. Mice that survived the first six weeks of life had neutralizing antibody to CTF virus. Neutralization tests, in which pregnant mice were inoculated with both CTF virus and specific antiserum to CTF virus, validated the teratogenic effects of CTF virus in mice.

Animals↗
PubMed1978

Colorado tick fever: clinical, epidemiologic, and laboratory aspects of 228 cases in Colorado in 1973-1974.

During 1973 and 1974, we looked for cases of Colorado tick fever throughout Colorado; 228 cases were identified. Although 90% of the patients reported exposure to ticks before illness, only 52% were aware of an actual tick bite. Typical symptoms of fever, myalgia, and headache were common, but gastrointestinal symptoms were also prominent in 20% of the patients. Twenty percent were hospitalized; no deaths or permanent sequelae were noted. Persistent viremia (greater than or equal to 4 weeks) was found in about half of the cases; this finding was not associated with the occurrence of prolonged symptoms (greater than or equal to 3 weeks), which were also reported in half of the cases. One patient became reinfected with the virus. Increasing tourism in endemic areas and the frequent occurrence of prolonged or biphasic illnesses provide the potential for patients with Colorado tick fever to seek medical care anywhere in the United States.

Adolescent↗