Edward M. Bosler

Spirochetes Isolated from the Blood of Two Patients with Lyme Disease

We isolated spirochetes from the blood of 2 of 36 patients in Long Island and Westchester County, New York, who had signs and symptoms suggestive of Lyme disease. The spirochetes were morphologically similar and serologically identical to organisms recently found to infect lxodes dammini ticks, which are endemic to the area and have been epidemiologically implicated as vectors of Lyme disease. In both patients, there was a rise in specific antispirochetal antibodies in paired specimens of serum. We conclude that the l. dammini spirochete has an etiologic role in Lyme disease.

Host Responses to Borrelia burgdorferi in Dogs and Horses

By using paired sera the IgM and IgG host responses were analyzed in dogs with ELISA and Western blot techniques. Antibodies in clinical seropositive dogs bound to 4-25 IgM and up to 40 or more IgG antigenic determinants. Early IgM response to the 41-kDa flagellin persisted for at least 9 months and involved as many as seven other peptides. IgG response expanded later in the disease and involved more immunogens than are currently recognized in late human disease. A percentage of asymptomatic dogs that later developed clinical symptoms were seropositive. Immunoblot studies suggested that B. burgdorferi is persistent in both asymptomatic and weakly reactive animals and if untreated could lead to disease expression. Clinical seropositive, asymptomatic seronegative, and experimentally infected horses were similarly studied. In experimentally inoculated animals IgG antibodies were initially bound to flagellin and later to the 34- and 31-kDa polypeptides, even though ELISA values were considered only slightly reactive.

Prevalence of canine lyme disease from an endemic area as determined by serosurvey

From August 1984 through February 1985, 423 dogs from 43 municipalities in 7 New Jersey counties were evaluated for the presence of antibodies to the Lyme disease spirochete (Borrelia burgdorferi). Of these dogs, 34.7% with no apparent clinical symptoms were serologically reactive (IFA greater than or equal to 1:64); titers in this study ranged from non-reactive to 1:2048. Ninety percent of the dogs surveyed had a current vaccination status to Leptospira interrogans serovars canicola and icterohaemorhagiae. Dogs vaccinated to leptospirosis elicited homologous antibody titers of less than or equal to 1:16 and, therefore, did not interfere with interpretation of antibody levels to B. burgdorferi. Effects of age, degree of outdoor activity, travel history, and location of residence were evaluated. The use of serosurveys of dogs as a tool for Lyme disease surveillance is discussed.

Comparison of rates of infection by the Lyme disease spirochete in selected populations of Ixodes dammini and Amblyomma americanum (Acari: Ixodidae)

At a major endemic focus in New Jersey, 50% of 290 adult Ixodes dammini collected in the fall of 1984 were infected with the Lyme disease spirochete (Borrelia burgdorferi), which was statistically higher than the rate found in the 202 adult ticks (39.6%) examined during the spring. Neither sex nor site of collection within the focus significantly affected the infection rate. The observed infection rates were similar to those reported in endemic areas of New York and Connecticut. Borrelia burgdorferi also infected all active stages of Amblyomma americanum ticks. Rates of infection were 5.4% in adults (n = 467) and 3.4% in nymphs (n = 289); 15.6% of clusters of unengorged larvae harbored B. burgdorferi, suggesting transovarial passage of the spirochete. Comparison of the rates of infection in I. dammini and A. americanum and their potential impact on Lyme disease transmission is discussed.

Human and Rodent Hantavirus Infection in New York State: Public Health Significance of an Emerging Infectious Disease

BACKGROUND A case of hantavirus pulmonary syndrome with possible exposure in New York and/or Rhode Island was confirmed in February 1994. OBJECTIVE To conduct four studies to determine the historical and geographic distribution of human and small-mammal infection with hantaviruses in New York State. METHODS Enzyme-linked immunosorbent assays were performed on serum samples obtained from 130 humans during a 1978 babesiosis survey, 907 small mammals collected in New York State since 1984, 12 rodents collected in 1994 near the residences of the patients with hantavirus pulmonary syndrome, and 76 New York patients with acute respiratory distress syndrome-like illness (as suspected cases of hantavirus pulmonary syndrome). RESULTS None of the human serum samples from the 1978 serosurvey showed evidence of hantavirus exposure by enzyme-linked immunosorbent assay. Statewide historical serum samples from white-footed mice showed evidence of Sin Nombre virus infection in 12.0% (97/809) and Seoul-like virus infection in 9.6% (78/809). Site-specific seropositivity rates were as high as 48.5% with Sin Nombre virus during 1 year (1984). Two of 12 mice captured near the residences of a human patient were positive for Sin Nombre virus by enzyme-linked immunosorbent assay, yet were negative for viral RNA by polymerase chain reaction. None of the patients with suspected hantavirus pulmonary syndrome was serologically reactive for Sin Nombre virus. CONCLUSIONS We provide serologic evidence of small-mammal infection with hantaviruses in New York State as long ago as 1984. Human cases of hantavirus pulmonary syndrome are rare in New York, and data indicate that transmission to humans is probably infrequent. A unique set of host, agent, and environmental factors may be necessary to cause hantavirus pulmonary syndrome in humans.

Evolution of a focus of Lyme disease

Epidemiological investigations were initiated in 1984 when significant Lyme disease activity was observed within a 5-km radius of an area previously used as a non-endemic control site for Lyme disease research in New Jersey. Through 1983, collections of Ixodes dammini from vegetation and feral rodents were infrequent and no human cases were identified within a 16-km radius of the control site. In 1984, 4 human cases and 3 serologically reactive canines (greater than or equal to 1:512) were recognized within the area and adult I. dammini populations were over 3-fold greater than those at our primary study location where Lyme disease has been endemic since 1981. Using darkfield microscopy, 53.4% of adult I. dammini were infected with Borrelia burgdorferi as compared to 50.0% of adults collected during the same period at the known endemic study site. These data indicate that a focus of Lyme disease has recently become established at the previously non-endemic control site and that the establishment of new foci may occur more rapidly than once thought.

The prevalence and significance of Borrelia burgdorferi in the urine of feral reservoir hosts.

Live Borrelia burgdorferi were isolated from the blood and/or urine of white-footed mice (Peromyscus leucopus) collected on Shelter Island, New York, in 1984 and 1985. Prevalence of spirochetes in urine was consistently higher than in blood or both fluids simultaneously. Spirochetes remained viable for 18-24 hours in urine and were maintained in culture for one week. Mice removed from the field were spirocheturic for at least 13 months. One spirocheturic mouse developed spirochetemia one month after field removal indicating the pathogen can return to the peripheral circulation. Twenty-one kidneys from 22 mice had spirochetes in the interstitial areas and bridging the tubules. A positive correlation between Babesia microti infection and spirocheturia was seen. Although the mechanism of entry into the urine is unknown, B. microti infection may increase glomerular permeability. Babesia induced hematuria may provide possible nutrients to maintain spirochetes. Urine may provide a method for contact non-tick transmission of B. burgdorferi in natural rodent populations particularly during periods of nesting and/or breeding.

Vector Tick Populations and Lyme Disease

Although many aspects of Lyme disease have been intensely studied for over a decade, little research has been directed toward control of the principal tick vector, Ixodes dammini. Ecological and epidemiological investigations have provided not only an ample understanding of tick biology and behavior, they have also identified the types of areas at risk for disease transmission. The advantages and limitations of previous attempts to control I. dammini by host reduction, habitat modification, and acaricide applications have been discussed in relation to overall control strategies for high-risk areas, and an integrated approach to control proposed.