Isolation of thogoto virus (Orthomyxoviridae) from the banded mongoose, Mongos mungo (Herpestidae), in Uganda.

Small wild vertebrates were trapped during an investigation into possible vertebrate reservoirs of o'nyong-nyong (ONN) fever virus in Uganda in 1997. Antibody neutralization test results and virus isolation attempts were negative for ONN virus, confirming the work of earlier investigators, who also failed to find evidence for a nonhuman ONN virus reservoir. In the course of these ONN virus studies, Thogoto virus was isolated from one of eight banded mongooses (Mongos mungo). This is the first isolation of Thogoto virus from a wild vertebrate. Neutralizing antibodies to Thogoto virus were also found in two of the other mongooses.

Isolation of Borrelia burgdorferi from Neotoma fuscipes, Peromyscus maniculatus, Peromyscus boylii, and Ixodes pacificus in Oregon.

The number of Lyme disease cases in Oregon has increased in recent years despite the fact that the pathogen, Borrelia burgdorferi, has never been isolated in the state. Rodent and tick surveys were undertaken in 1997 to isolate and characterize strains of B. burgdorferi from Oregon and to identify potential reservoirs and vectors of Lyme disease. Borrelia burgdorferi was isolated from Neotoma fuscipes, Peromyscus maniculatus, P. boylii, and Ixodes pacificus. Both N. fuscipes and P. maniculatus were infested with I. pacificus and I. spinipalpis. Although I. pacificus infested P. boylii, I. spinipalpis was not found on this rodent, and only 4% of the P. boylii were infected with B. burgdorferi compared with the 19% and 18% infection rates found in N. fuscipes and P. maniculatus, respectively. Variation in the molecular weights of the outer surface proteins A and B were found in these first confirmed isolates of B. burgdorferi from Oregon, as well as truncated forms of outer surface protein B.

Amblyomma americanum: a potential vector of human ehrlichiosis.

Polymerase chain reaction primers specific for Ehrlichia chaffeensis were used to amplify DNA from extracts of pooled ticks. Amplification was performed on extracts from 140 pools (1,579 total ticks) consisting of three tick genera collected from five states. The characteristic 389-basepair product was observed after amplification of extracts from seven different pools of adult Amblyomma americanum (117 pools, 1,462 ticks), but not from pools of nymphs. No specific product was observed after amplification of 20 pools (105 ticks) of Dermacentor variabilis and three pools of Ixodes scapularis (12 ticks). Ehrlichia chaffeensis was present in A. americanum at a minimum frequency of > or = 0.48%, suggesting that A. americanum may be a vector of human ehrlichiosis.

Use of a sentinel host system to study the questing behavior of Ixodes spinipalpis and its role in the transmission of Borrelia bissettii, human granulocytic ehrlichiosis, and Babesia microti.

Ixodes spinipalpis maintains Borrelia bissettii spirochetes in Colorado in a cycle involving wood rats and deer mice. This tick has been described as nidicolous, remaining either attached to its rodent hosts or in the rodent nest. Nidicolous ticks pose little risk of pathogen transmission to humans if they do not actively quest for hosts. To investigate the questing potential of I. spinipalpis, sentinel mice were placed in an area where I. spinipalpis had been commonly found on wood rats and deer mice. Concurrently, wild rodent populations were trapped and analyzed for Lyme disease spirochetes, the agent of human granulocytic ehrlichiosis (aoHGE), and Babesia microti. A total of 122 I. spinipalpis larvae and 10 nymphs were found on 19% of 244 sentinel mice. In addition, 4 sentinel mice became infested with Malaraeus telchinus or Orchopeas neotomae fleas. Questing I. spinipalpis were positively associated with woody shrubs and negatively associated with sunny and grassy areas. Four sentinel mice became infected with aoHGE after having been fed upon only by I. spinipalpis larvae. One sentinel mouse became infected with B. bissettii after having an I. spinipalpis nymph feed on it, and one sentinel mouse became coinfected with aoHGE and B. bissettii after it was fed upon by a single I. spinipalpis nymph. These sentinel mouse conversions suggest the possibility that the aoHGE is transovarially transmitted by I. spinipalpis, and that I. spinipalpis is capable of simultaneously transmitting B. bissettii and the aoHGE. The findings that I. spinipalpis quest away from rodent nests and will attach to and infect sentinel mice may be of public health importance. It suggests the potential transmission of the agents of human granulocytic ehrlichiosis and Lyme disease to other hosts by I. spinipalpis, in regions of the western United States where Ixodes pacificus is not found.

Distribution, diversity, and host specificity of Bartonella in rodents from the Southeastern United States.

A number of Bartonella isolates were obtained from seven species of rodents sampled from 12 geographic sites representing the major biotic communities of the southeastern United States. Bartonella were isolated from the blood of 42.2% of 279 tested rodents. The highest prevalence of infection typically occurred among the most commonly captured species in the rodent community. Four phylogenetic groups, uniting 14 genotypic variants of Bartonella, were identified by sequence analysis of the citrate synthase gene. The level of sequence homology between genotypic groups varied from 88.8% to 96.4%, and the degree of homology among variants within groups was > or = 97%. Cotton rats (Sigmodon hispidus) harbored up to three phylogenetic groups of Bartonella at a single site, and Bartonella of two phylogenetic groups were isolated from a single rodent. All the Bartonella isolated from three species of Peromyscus clustered in a single distinct phylogenetic group, suggesting some host specificity may occur. Mouse ascitic fluids produced in BALB/c mice inoculated with Bartonella of three phylogenetic groups demonstrated high indirect fluorescent antibody (IFA) titers to homologous antigens. However, use of eight Bartonella antigens in an IFA test with sera from 394 wild-caught rodents resulted in either little or extremely low titers of antibody.

Population genetics and phylogenetic analysis of Colorado Borrelia burgdorferi.

Borrelia burgdorferi is transmitted in an enzootic cycle in Colorado between the tick Ixodes spinipalpis and the woodrat Neotoma mexicana. The genetic relationship of Colorado isolates to other B. burgdorferi isolates is unknown nor have relationships among various Colorado isolates been determined. Portions of the flagellin (fla), 66-kD protein, and outer surface protein A (ospA) genes were amplified from 71 Colorado isolates, screened for genetic variability using single strand conformation polymorphism analysis, and unique alleles were sequenced. Colorado isolates were most similar to tick isolates from California and New York isolate 25015. Genetic distances among Colorado ospA sequences were the same or higher than distances among other isolates whereas distances among fla sequences tended to be the same or lower. The index of association (I(A)) was calculated among all loci as a measure of clonality. The I(A) among Colorado isolates was similar to I(A) previously estimated among other United States isolates.

Molecular investigation of a multisource outbreak of Crimean-Congo hemorrhagic fever in the United Arab Emirates.

During the investigation of an outbreak of Crimean-Congo hemorrhagic fever (CCHF) in the United Arab Emirates (UAE) between 1994 and 1995, blood samples from suspected CCHF cases and ticks collected from livestock were tested for CCHF virus by antigen-capture ELISA and by a reverse transcription-polymerase chain reaction. Phylogenetic analysis of partial small (S) segment nucleotide sequences from four ticks and five human samples showed that with one exception, all the human and tick viruses clustered along with samples from Pakistan and Madagascar in one distinct lineage. Within this lineage, sequences from the UAE patients were identical or closely related to those from three Hyalomma spp. ticks obtained from livestock recently imported from Somalia. Another sequence from a UAE patient was more closely related to a CCHF virus from Nigeria. These data indicate that the 1994-1995 CCHF epidemic in the UAE was a multisource outbreak possibly associated with importation of CCHF virus-infected livestock and ticks.

An outbreak of Crimean-Congo hemorrhagic fever in the United Arab Emirates, 1994-1995.

A multi-faceted investigation was conducted in the United Arab Emirates to characterize the epidemiologic and ecologic factors underlying an outbreak of Crimean-Congo hemorrhagic fever (CCHF) noted in November 1994 among abattoir workers. A chart review was conducted among hospitalized suspected cases of viral hemorrhagic fever with onset between January 1994 and March 1995 coupled with serologic testing of available specimens for the presence of virus antigen and IgG and IgM antibodies by ELISA. Livestock handlers and animal skin processors were interviewed and tested for the presence of IgG antibody. Sera from imported and domestic ruminants were examined for antibody for CCHF virus, and ticks collected from these animals were tested with an antigen-capture ELISA. Thirty-five suspected cases of CCHF were identified (case fatality = 62%). Livestock market employees, abattoir workers, and animal skin processors accounted for 16 (57%) of 28 cases with known occupational status. Serologic evidence of past asymptomatic infection was noted in 12 (4%) of 291 livestock and abattoir workers but in none of the controls. Nineteen (7%) of 268 animals were positive for CCHF virus antibodies by ELISA including 12 ruminants from Somalia and Iran and five indigenous camels. One Hyalomma impeltatum and two H. excavatum from Somali cattle and one H. anatolicum from a Somali goat were positive for CCHF virus antigen.

Prevalence of antibodies to arenaviruses in rodents from the southern and western United States: evidence for an arenavirus associated with the genus Neotoma.

The objectives of this study were to extend our knowledge of the geographic distribution and rodent host range of arenaviruses in North America. Sera from wild rodents collected from the southern and western United States were tested for antibody against Tamiami, Pichinde, Junin, and lymphocytic choriomeningitis viruses, using an indirect fluorescent antibody test. Antibody to at least one arenavirus was found in 220 (3.1%) of 7,106 rodents tested. The antibody-positive animals included Mus musculus from Florida and Texas; Neotoma albigula from Arizona, Colorado, and New Mexico; N. fuscipes and N. lepida from California: N. mexicana from Arizona, New Mexico, and Utah; N. stephensi from Arizona and New Mexico; and Oryzomys palustris and Sigmodon hispidus from Florida. Sigmodon hispidus seropositive for Tamiami virus were found only in Florida (156 [27.0%] of 578 tested), although 463 hispid cotton rats from outside that state were examined. High-titered antibodies to Tamiami virus were present in sera from S. hispidus, (geometric mean antibody titer [GMAT] of 1:792), whereas sera from Neotoma spp. reacted at high titer to both Tamiami (GMAT = 1:905) and Pichinde (GMAT = 1:433) viruses. The results suggest that arenaviruses are widely distributed in the southern United States and that one or more indigenous arenaviruses are associated with Neotoma spp. in North America.

A household-based, case-control study of environmental factors associated with hantavirus pulmonary syndrome in the southwestern United States.

During an outbreak of hantavirus pulmonary syndrome (HPS) in the southwestern United States, trained environmental assessment teams conducted surveys at 17 case-patient homes and matched controls from June through August 1993. Variables related to rodent abundance were quantified and standardized rodent trapping was conducted around and within households. The majority of households were located in pinon-juniper vegetation zones, and there were no significant differences in the type of house in which cases and controls lived. The only environmental factor that distinguished case households from controls was significantly higher small rodent densities (median trap success for case sites = 17.3%, 12.7% for near controls, and 8.3% for far controls). Frequency of hantaviral infection in deer mice (Peromyscus maniculatus) did not vary significantly among households of cases and controls, with a range of 27.5-32.5% antibody-positive. Indices of rodent fecal contamination were slightly higher in case houses. The data indicate that higher rodent densities were associated with households in which HPS cases occurred. Strategies that control rodent numbers and decrease rodent access to dwellings may reduce risk of human infection.

Acaricide susceptibility of immature Ixodes scapularis (Acari: Ixodidae) as determined by the disposable pipet method.

The disposable pipet method was used to establish baseline dose-mortality data for immature stages of Ixodes scapularis Say for carbaryl, esfenvalerate, cyfluthrin, and permethrin. Permethrin and cyfluthrin were most toxic to both nymphs and larvae, followed in order by esfenvalerate and carbaryl, based on LD50 and LD90 values. Larvae were substantially more susceptible than nymphs to each compound.

Studies on the control of plague in the western United States: laboratory trials of six insecticide dust formulations applied to soil for the control of the plague vector Oropsylla montana (Siphonaptera: Ceratophyllidae).

In this study, six insecticide dust formulations were laboratory-tested in soil mixtures ranging in concentration from 40 to 1,280 parts per million (ppm) against a proven vector of bubonic plague, Oropsylla montana (Baker). Pulicidal effects of the six insecticides, which included three organophosphates, two carbamates, and a pyrethroid, were compared by LD(50) values at both ambient and 75% RH. Permethrin (Pyraperm 455 dust), a pyrethroid with a LD(50) less than 40 ppm, demonstrated the most effective pulicidal action for the 13-wk trial period, followed in order by chlorpyrifos, bendiocarb, propetamphos, diazinon, and carbaryl.

Reported cases of human plague infections in the United States, 1970-1991.

From 1970 to 1991, 295 indigenous cases and one imported case of human plague were reported in the United States. Eighty-two percent of the total indigenous cases occurred in New Mexico, Arizona, and Colorado. Ninety-three percent of these cases had onset in the months of April through November. Most cases (89%) presented as bubonic or septicemic plague, or both. Cases were reported more frequently in males (58%), and male mortality exceeded that of females (17 versus 11%). Ground squirrels were the most frequently implicated sources of infection in cases associated with flea bites, and domestic cats were found to play an increasingly important role in transmission of disease to humans during these decades.

Laboratory and field trials of permethrin-treated cotton used as nesting material to control fleas (Insecta: Siphonaptera) on cricetid rodents.

Upholstery cotton treated with four different concentrations (0.25-2.0%) (2,500-20,000 ppm) of an aqueous permethrin solution, used as nesting material by white mice, was laboratory-tested against the potential plague vectors Oropsylla montana (Baker), Thrassis bacchi (Rothschild), and Orchopeas howardi (Baker) and found highly effective (P less than 0.001) for 1 yr. Similarly treated cotton gauze was tested under ambient and 75% RH and was found to be highly effective (P less than 0.001) in both environments for 1 yr. A separate test determined that the LD50 of permethrin-treated cotton was less than 10 ppm. Cotton tested with 0.5% permethrin and distributed under field conditions to cricetid rodents for use as nesting material was found to be highly effective (P less than 0.001 as a pulicide for greater than 4 mo when tested during winter in Larimer County, Colo. Permethrin-treated cotton was less successful in controlling fleas on cricetid rodents during the summer months in a New Mexico hyperendemic plague area.

Infection of Ixodes scapularis (Acari: Ixodidae) with Borrelia burgdorferi using a new artificial feeding technique.

To study interactions between Ixodes scapularis (Say) and Borrelia burgdorferi, an artificial feeding system was refined to allow controlled manipulation of single variables. The feeding system uses a mouse skin mounted on a water-jacketed glass membrane feeder. I. scapularis were infected using either BSK-H-cultured B. burgdorferi spirochetes or a B. burgdorferi-infected mouse skin as the source of spirochetes. Sixty-six percent of nymphs successfully fed to repletion using the artificial feeding systems with at least 75% of nymphs becoming infected with B. burgdorferi. Strain B31 B. burgdorferi spirochetes from passages 2-17 were equally infectious to nymphal ticks. At concentrations of one spirochete per microliter, 12% of nymphs acquired infection and 14 and 100 spirochetes per microliter resulted in 50 and 100% infection rates, respectively. Eighty-nine percent of nymphs fed by artificial feeding molted to the adult stage. When subsequently fed as adults, these I. scapularis successfully transmitted infectious B. burgdorferi spirochetes to mice.

Vector competence of Ixodes scapularis, I. spinipalpis, and Dermacentor andersoni (Acari:Ixodidae) in transmitting Borrelia burgdorferi, the etiologic agent of Lyme disease.

This report describes the vector competence of 3 ixodid tick species, Ixodes scapularis (Say), I. spinipalpis (Nuttall), and Dermacentor andersoni (Stiles), for Borrelia burgdorferi in Colorado. The study was based on preliminary field work performed in 6 Colorado counties, where rodents and ticks were collected and assayed for the presence of B. burgdorferi. Four of the 6 counties produced 52 rodent and 39 I. spinipalpis isolates of B. burgdorferi. Two B. burgdorferi isolates were tested under laboratory conditions and found to be infective to Imperial Cancer Research Fund (ICRF) outbred mice. The 1st, a low-passage strain originating from New York (B-31, passage 6) was used as a control, and the 2nd was isolated from ear tissue of a Neotoma mexicana (Baird) (Mexican wood rat) that was trapped in Colorado. Larvae of I. scapularis, I. spinipalpis. and D. andersoni were fed on infected mice and cultured in Barbour-Stoner-Kelly media to assay for infection at 1, 2, 3, and 4 wk after repletion. The infection rates in replete larvae. were 75, 69, and 8.5%, respectively, whereas transstadial nymphal infection rates were 80, 75, and 0%, respectively. Both I. scapularis and I. spinipalpis were shown to be competent vectors that acquired the infection from the host reservoir mice and subsequently transmitted it to naive mice. Given that I. scapularis are not found in Colorado, I. spinipalpis are restricted to the nests and burrows of rodents, and because of the semiarid environment in Colorado, the risk of human contact with B. burgdorferi appears to be low.

Flea (Siphonaptera:Ceratophyllidae, hystrichopsyllidae) and tick (Acarina:Ixodidae) control on wood rats using host-targeted liquid permethrin in bait tubes.

The efficacy of a liquid permethrin-treated bait tube controlling fleas and ticks on Mexican wood rats, Neotoma mexicana Baird, was evaluated during a 1-yr study in north-central Colorado. Results indicated that the bait tubes were effective for reducing flea and tick infestations on wood rats. The effects of treatment persisted throughout the study, despite the fact that bait tubes were replenished with bait and permethrin only during the first 4 mo (4 replenishments). Our results suggest that these bait tubes provide an effective, economical, and environmentally acceptable means of controlling vectors of flea or tick-borne diseases, although slight modifications of the basic bait tube design might be required to maintain the effectiveness of the tube under different ecological conditions.

Susceptibility of immature Ixodes scapularis (Acari:Ixodidae) to plant-derived acaricides.

Plant-derived acaricides, extracted from various botanical species, and commercially available phytochemicals were evaluated for biological activity against immature Ixodes scapularis (Say) using the disposable pipet method. In addition, residual activity of the plant extracts was determined. Of the 13 plant extracts tested, 9 exhibited biological activity with Alaska yellow cedar, Chamaecyparis nootkatensis (D. Don) Spach., being the most effective against the nymphal ticks (LC50 = 0.151% wt:vol) and eastern red cedar, Juniperus virginiana L., showing the greatest activity against larval ticks (LC50 = .001% wt:vol). The commercially available products were significantly less active than the plant extracts we prepared, but some commercial compounds did exhibit limited activity. Only the Alaska yellow cedar exhibited any residual activity that lasted 21 d after treatment.

Vector competence of Ixodes scapularis and Ixodes ricinus (Acari: Ixodidae) for three genospecies of Borrelia burgdorferi.

The vector competence of 2 tick species, Ixodes ricinus (L.) and Ixodes scapularis Say, was determined and compared for 3 genospecies of Borrelia burgdorferi. The 3 genospecies of B. burgdorferi used in the following experiments were Borrelia burgdorferi sensu stricto (B-31 and B-31.D1 clone), Borrelia afzelii (strain Pgau. C3), and Borrelia garinii (strain VS286 and VSBP). Spirochetes from all 5 strains were inoculated intradermally into outbred mice; larval ticks of both species were subsequently fed on those mice and replete larvae were assayed for infection by culture in BSK-H media every 7 d for 4 wk. Infection frequencies in I. scapularis exposed to the 5 strains were as follows: B-31 (90%), B-31.D1 (83%), Pgau.C3 (87%), VS286 (10%), and VSBP (5%). The comparable infection frequencies for I. ricinus were B-31 (3%), B-31.D1 (3%), Pgau.C3 (90%), VS286 (5%), and VSBP (3%). Resultant nymphal I. scapularis successfully transmitted B-31, B-31,D1, Pgau.C3, and VS286 to outbred mice. I. ricinus nymphs transmitted Pgau.C3 and VS286. Both species failed to transmit strain VSBP.

Plague (Yersinia pestis) in cats: description of experimentally induced disease.

Sixteen healthy cats were fed a 6-wk-old laboratory mouse that had died of experimentally induced Yersinia pestis infection (strain NM77-538), to simulate oral exposure to plague. The cats were closely monitored after ingestion. Physical exams were performed and vital signs were recorded daily. Plague antibody titers and cultures of blood, throat, and oral cavity were performed daily. Complete blood counts and biochemistry panels were performed every 3 d. Complete necropsies were performed on any cats that died. Cats exhibited one of three responses following ingestion of one plague-infected mouse; they either died (6/16 or 38%), developed transient illness and recovered (7/16 or 44%) or showed no signs of illness (3/16 or 19%). A continual fever greater > 40 degrees C was associated with a poor prognosis. The highest antibody titers developed in the group that shed the plague bacillus over an extended period of time. Blood, throat, and oral cavity cultures were positive in 100% of the fatal cases. Throat cultures were positive in 75% of the exposed cats. In contrast to other carnivores, cats infected with Y. pestis exhibit bubo formation and pneumonic lesions similar to those seen in people with plague. Because of the potential transmission of Y. pestis from cats to people, development of a plague vaccine for cats may be warranted.