West Nile virus isolates from mosquitoes in New York and New Jersey, 1999.

An outbreak of encephalitis due to West Nile (WN) virus occurred in New York City and the surrounding areas during 1999. Mosquitoes were collected as part of a comprehensive surveillance program implemented to monitor the outbreak. More than 32,000 mosquitoes representing 24 species were tested, and 15 WN virus isolates were obtained. Molecular techniques were used to identify the species represented in the WN virus-positive mosquito pools. Most isolates were from pools containing Culex pipiens mosquitoes, but several pools contained two or more Culex species.

West Nile virus in overwintering Culex mosquitoes, New York City, 2000.

After the 1999 West Nile (WN) encephalitis outbreak in New York, 2,300 overwintering adult mosquitoes were tested for WN virus by cell culture and reverse transcriptase-polymerase chain reaction. WN viral RNA and live virus were found in pools of Culex mosquitoes. Persistence in overwintering Cx. pipiens may be important in the maintenance of WN virus in the northeastern United States.

Rapid detection of west nile virus from human clinical specimens, field-collected mosquitoes, and avian samples by a TaqMan reverse transcriptase-PCR assay.

The authors report on the development and application of a rapid TaqMan assay for the detection of West Nile (WN) virus in a variety of human clinical specimens and field-collected specimens. Oligonucleotide primers and FAM- and TAMRA-labeled WN virus-specific probes were designed by using the nucleotide sequence of the New York 1999 WN virus isolate. The TaqMan assay was compared to a traditional reverse transcriptase (RT)-PCR assay and to virus isolation in Vero cells with a large number ( approximately 500) of specimens obtained from humans (serum, cerebrospinal fluid, and brain tissue), field-collected mosquitoes, and avian tissue samples. The TaqMan assay was specific for WN virus and demonstrated a greater sensitivity than the traditional RT-PCR method and correctly identified WN virus in 100% of the culture-positive mosquito pools and 98% of the culture-positive avian tissue samples. The assay should be of utility in the diagnostic laboratory to complement existing human diagnostic testing and as a tool to conduct WN virus surveillance in the United States.

First field evidence for natural vertical transmission of West Nile virus in Culex univittatus complex mosquitoes from Rift Valley province, Kenya.

West Nile virus is a mosquito borne flavivirus endemic over a large geographic area including Africa, Asia, and the Middle East. Although the virus generally causes a mild, self-limiting febrile illness in humans, it has sporadically caused central nervous system infections during epidemics. An isolate of West Nile virus was obtained from a pool of four male Culex univittatus complex mosquitoes while we were conducting an investigation of Rift Valley fever along the Kenya-Uganda border in February-March 1998. This represents the first field isolation of West Nile virus from male mosquitoes and strongly suggests that vertical transmission of the virus occurs in the primary maintenance mosquito vector in Kenya. A phylogenetic analysis of the complete amino acid sequence of the viral envelope glycoprotein demonstrated a sister relationship with a Culex pipiens mosquito isolate from Romania made in 1996. This unexpected finding probably reflects the role of migratory birds in disseminating West Nile virus between Africa and Europe.

La Crosse viremias in white-tailed deer and chipmunks exposed by injection or mosquito bite.

To further understand the role of wild mammals in the maintenance of La Crosse virus (LACV) in nature, we investigated the effects of inoculation method and virus source on the duration and amplitude of LACV viremia in vertebrate hosts. Earlier work suggested that deer are not sufficiently susceptible to LACV to play an important role in its maintenance. We re-evaluated the susceptibility of deer since subsequent studies showed that they constitute 65% of Aedes triseriatus blood meals, and thus would be exposed frequently to the virus. In our study, deer developed higher and longer viremia following exposure to LACV by infected Ae. triseriatus than those previously reported by inoculation with needle and syringe. However, susceptible Ae. triseriatus that fed on these viremic animals did not become infected. Because a large number of uninfected mosquitoes can feed upon a viremic deer in nature, we believe that deer should not be disregarded completely as a possible amplifier in the LACV transmission cycle. We also infected chipmunks to determine if there were significant differences in viremia response from mosquito delivery of virus to the chipmunk host, compared with artificial exposure by injection. Chipmunks exposed to infected mosquitoes had higher and longer viremias than the ones produced by intramuscular injection of an LACV suspension. These findings show the importance of using LACV infected mosquitoes for transmission experiments in mammals.

California serogroup virus infections in Wisconsin domestic animals.

A serologic survey and experimental virus transmission studies were done to assess the role of domestic animals as amplifier hosts of La Crosse (LACV) and Jamestown Canyon (JCV) viruses. Serum from 319 cows, 88 dogs, 122 equines, 47 swine, 10 goats, and 4 cats were tested for neutralizing antibody to LACV, JCV, trivittatus (TVTV), and snowshoe hare (SSHV) viruses. Antibody prevalences of LACV, TVTV, and SSHV were less than 10% in all species. Antibody to JCV was detected in all species except cats. Prevalence ranged from 10% in goats and swine to 29% in dogs. No age-associated trends in JCV prevalence were noted. Two of 6 adult dogs, and 2 of 4 pigs inoculated with 6.3-6.5 log10 suckling mouse intracerebral 50% lethal doses (SMICLD50) of LACV developed viremias ranging of less than 1.0-2.9 log10 SMICLD50/ml 1-3 days after inoculation. Of 4 puppies inoculated with LACV, 3 developed fatal infections. Viremias were not detected in 4 cows, 4 ponies, 7 cats, or 6 sheep. Two cats fed LACV infected suckling mice shed virus from the oropharynx for 1 day each. All animals except 1 cow, 1 cat, and 1 sheep had greater than or equal to 4-fold rise in antibody titers. Five additional dogs fed upon by LACV-infected Aedes triseriatus mosquitoes did not develop viremias or antibody and uninfected Ae. triseriatus engorging on the dogs 1-5 days after feeding by infected mosquitoes failed to become infected. Five ponies, 6 calves, 2 ewes, 6 dogs, and 5 piglets were inoculated with 3.6-7.3 log10 SMICLD50 of JCV. None developed detectable viremias, although greater than or equal to 4-fold rises in antibody titers developed in 60% of the ponies, 17% of the calves, 50% of the dogs, and 1 of 2 ewes. None of the pigs developed corresponding rises in antibody titers. We conclude that juvenile and adult animals of the species tested are not efficient amplifier hosts of LACV or JCV, but may be useful sentinels of local virus transmission.

Lyme disease ecology in Wisconsin: distribution and host preferences of Ixodes dammini, and prevalence of antibody to Borrelia burgdorferi in small mammals.

Lyme disease recently has been recognized in Wisconsin. Trapping studies were conducted at four geographically separate and ecologically distinct regions in Wisconsin to elucidate the distribution and host preferences of Ixodes dammini on small and medium sized mammals, and the occurrence of antibodies to Borrelia burgdorferi in these wild mammals. Peak I. dammini larval activity occurred from June-September. Nymphs were most active from May-August. White-footed mice (Peromyscus leucopus) and chipmunks (Tamias striatus) were important hosts for immature ticks. Mean numbers of I. dammini per mouse were highest in regions of high prevalence of Lyme disease. Antibody to B. burgdorferi was detected in sera of 60/371 (16%) white-footed mice, 5/104 (5%) chipmunks, 3/5 (60%) gray squirrels (Sciurus carolinensis), 0/8 raccoons (procyon lotor), and 0/12 opossum (Didelphis virginiana); antibody prevalence correlated positively with I. dammini occurrence, and seropositive animals were not detected in areas where I. dammini were not found. Two of 15 recaptured P. leucopus had greater than or equal to 4-fold changes in antibody titer. B. burgdorferi was cultured from blood of a P. leucopus captured in west-central Wisconsin, and was observed by direct immunofluorescence in 9/23 (39%) I. dammini nymphs. In Wisconsin, I. dammini has increased in numbers and has significantly expanded its range since its first recognition in 1968.

Babesiosis in Wisconsin. A new focus of disease transmission.

A confirmed case of human babesiosis was identified in August 1983 in a 54-year-old asplenic Wisconsin resident. Babesia microti was identified as the causative agent by blood smear morphology and hamster inoculation techniques. The patient's wife had clinically confirmed Lyme disease in 1981 and had serologic evidence (immunofluorescent antibody to a B microti titer of 1:1,024) of recent Babesia infection in August 1983. Mice (Peromyscus species) trapped on the patients' property and elsewhere in their Wisconsin county of residence were infected with B microti. Lyme disease and babesiosis have the same tick vector and animal reservoir; serum samples from 116 Wisconsin and Minnesota residents with clinically confirmed Lyme disease between 1980 and 1983 were tested, and none were found to have concurrent Babesia infection. This area of Wisconsin is identified as a new focus for babesiosis transmission, but the risk of transmission seems to be low.

Lyme disease in Wisconsin: epidemiologic, clinical, serologic, and entomologic findings.

In 1980-82, 80 individuals (71 Wisconsin residents) had confirmed Lyme disease (LD-c) reported; 39 additional patients had probable or possible LD. All cases of LD-c occurred during May-November; 73 percent occurred during June-July; 54 (68 percent) occurred in males. The mean age was 38.7 years (range, 7-77 years). Among LD-c patients, likely exposure to the presumed vector Ixodes dammini (ID) occurred in 22 different Wisconsin counties. Antibodies to the ID spirochete that causes LD occurred in 33 of 49 LD-c cases versus 0 of 18 in ill controls (p less than .001) and in 13 of 26 LD-c cases treated with penicillin or tetracycline versus 16 of 19 LD-c cases not treated. Early antibiotic therapy appears to blunt the antibody response to the ID spirochete. Regional tick surveys conducted in Wisconsin during each November in 1979-82 have demonstrated regions of greater density of ID. Utilizing comparable tick collection in these surveys, increases were noted in the percentage of deer with ID from 24 percent (31/128) in 1979 to 38 percent (58/152) in 1981, in the standardized mean value of ID/deer from 1.0 in 1979 to 2.2 in 1981, in the percentage of ID of the total ticks collected from 13 percent in 1979 to 71 percent in 1981, or in the ratio of ID to Dermacentor albipictus ticks from 0.14 in 1979 to 2.44 in 1981. However, a reduction in the density of ID/deer was noted generally throughout Wisconsin in 1982 when compared to 1981. LD is widespread in Wisconsin, with ecologic and clinical features similar to those occurring along the eastern seaboard.