Transmission of Anaplasma phagocytophila (human granulocytic ehrlichiosis agent) in horses using experimentally infected ticks (Ixodes scapularis).

Most human granulocytic ehrlichiosis (HGE) studies carried out in horses use needle inoculation of infected leucocytes or cell cultures. This route of inoculation does not accurately reflect natural infection of the tick-borne agent. To investigate whether tick transmission influences the course of granulocytic ehrlichiosis in the horse model, experimental transmission through infected laboratory-reared Ixodes scapularis ticks was attempted into two healthy horses. One additional horse served as negative control and was exposed to uninfected ticks. Eleven days after exposure to nymphal or adult ticks infected with Anaplasma phagocytophila (HGE agent) the two horses developed severe clinical and laboratory signs consistent with granulocytic ehrlichiosis. Bacteraemia was determined at various time points in the two horses by observation of morulae within neutrophils and by detection of A. phagocytophila genomic DNA by PCR of peripheral blood leucocytes. Further, both horses seroconverted. In contrast the control horse stayed uninfected. The results demonstrate that A. phagocytophila can be experimentally transmitted by infected nymphal and adult ticks and that the agent is able to produce a severe disease, similar to naturally occurring cases. Therefore, tick transmission is highly reproducible and can be successfully used in the equine animal model in order to study HGE.

Ectoparasites of the island fox on Santa Cruz Island.

The ectoparasite fauna for island foxes (Urocyon littoralis) on Santa Cruz Island (California, USA) in April (wet season) and September (dry season) 1998 was evaluated. Three taxa of ectoparasites were identified. These were fleas (Pulex irritans), lice (Neotrichodectes mephitidis), and ticks (Ixodes pacificus). Ectoparasite abundances varied seasonally. Typical of insular endemic species, island foxes may be especially vulnerable to the introduction of novel disease organisms and their vectors.

Quantitative evaluation of ehrlichial burden in horses after experimental transmission of human granulocytic Ehrlichia agent by intravenous inoculation with infected leukocytes and by infected ticks.

This paper describes the kinetics of the human granulocytic ehrlichiosis agent in the blood of horses experimentally infected by intravenous inoculation with infected leukocytes and by infected ticks as evaluated by using a real-time quantitative PCR assay. The data obtained indicated differences in the period of incubation, duration of rickettsemia, and initial and maximal ehrlichial loads between the two routes of infection.

Association of Ixodes pacificus (Acari: ixodidae) with the spatial and temporal distribution of equine granulocytic ehrlichiosis in California.

This study was conducted to determine if the biology of certain ticks associated with horses regulates the spatial and temporal distribution of equine granulocytic ehrlichiosis (EGE) in California north of Monterey County. We compared the spatial and temporal distribution of EGE cases with the seasons of activity and life histories of ticks that infest horses. Spatially, cases collected from equine veterinarians clustered around each other in a manner different from the way in which control cities of practice were distributed, with foci limited to the Sierra Nevada and coastal foothills. Cases also clustered seasonally: most were diagnosed between November and April. The spatial and temporal pattern of EGE cases closely parallels the well-characterized life history and distribution of Ixodes pacificus Cooley & Kohls, but not other ticks commonly associated with horses. Building on previous studies, there is compelling evidence that this tick has the vectorial capacity to transmit Ehrlichia equi to horses. Based on the life history and distribution of I. pacificus in relation to EGE cases, we reason that this tick is the only biologically plausible vector of E. equi in California, and provide evidence for a tightly linked association between I. pacificus and the epidemiology of EGE.

Ehrlichia spp. in cervids from California.

Blood samples from six mule deer (Odocoileus hemionus hemionus), 15 black-tailed deer (O. hemionus columbianus), and 29 elk (Cervus elaphus nannodes) were assayed for human monocytic and human granulocytic ehrlichiosis (HGE) by polymerase chain reaction (PCR), DNA sequencing, and serology to determine whether or not cervids are involved in the maintenance of these potential human pathogens in California (USA). The deer were sampled in August to October 1992-95. The 29 tule elk from Point Reyes National Seashore were sampled in August 1997. All deer were seronegative for antibodies to HGE/Ehrlichia equi, while the E. equi seroprevalence among elk was 17%. The 16S rDNA PCR prevalence in deer was 38% (in mule deer and black-tailed deer) for Ehrlichia-like sp. of white-tailed deer, 5% (one black-tailed deer only) for E. equi, and 0% for E. chaffeensis. The PCR prevalence in elk was 0% for Ehrlichia-like sp. of white-tailed deer, 31% for E. equi, and 0% for E. chaffeensis. The E. equi from two positive elk samples was successfully propagated in HL-60 cell cultures. DNA sequencing confirmed that the Ehrlichia-like sp. sequences from deer in California were closely related to sequences reported from white-tailed deer from Oklahoma and Georgia. The E. equi strain from deer and elk resembled other E. equi strains from California. These results suggest that cervids may be important in the natural maintenance of E. equi in California.

Experimental transmission of Ehrlichia equi to horses through naturally infected ticks (Ixodes pacificus) from Northern California.

We report the experimental transmission of Ehrlichia equi from naturally infected Ixodes pacificus ticks to horses. Three weeks after exposure to ticks, two of three horses developed clinical signs compatible with E. equi infection, while one horse remained asymptomatic. 16S rRNA gene PCR of blood leukocyte lysates was positive for all horses at various time points; two horses seroconverted. The 16S rRNA gene sequences amplified from tick-exposed horses showed more than 99% homology to corresponding fragments of the 16S rRNA genes of E. equi, Ehrlichia phagocytophila, and the human granulocytic ehrlichiosis agent.

Ranging of male Oropsylla montana fleas via male California ground squirrel (Spermophilus beecheyi) juveniles.

We asked if fleas more frequently remain on those California ground squirrels (Spermophilus beecheyi) that are likely to emigrate from their natal nest. In Camp Ohlone, Alameda County, California, juvenile male squirrels were infested with more fleas (Oropsylla montana) than were juvenile females, and juveniles of both sexes were infested with more fleas (O. montana and Hoplopsyllus anomalus) than adults. There was no difference between the adult sexes in the number of fleas. The disproportionate infestation of male juveniles was accounted for almost exclusively by male O. montana. Greater activity on the part of juvenile males did not account for this difference; the activity of male and female juveniles was very similar. As yearlings, male squirrels established home ranges at greater distances from the natal burrow than did females. Remaining on ranging male squirrels might help male fleas find nonsibling mates in new nests, whereas female fleas might tend to stay in the natal nest in order to assure their progeny of its resources. Flea behavior, modified by characteristics of the host that are sex-specific and predictive of future traits, such as the tendency to range, may thus determine the nature and extent of infestations in juvenile squirrels.

Ixodes pacificus (Acari: Ixodidae) as a vector of Ehrlichia equi (Rickettsiales: Ehrlichieae).

Ehrlichia equi, a rickettsia described from horses in California 30 yr ago, causes equine granulocytic ehrlichiosis throughout the Americas and possibly Europe. Here, we report experimental transmission of E. equi from infected to susceptible horses through bites of western blacklegged ticks, Ixodes pacificus (Cooley & Kohls). In preliminary field studies, only I. pacificus consistently infested horses and vegetation at 3 locations with contemporary cases of equine ehrlichosis, and in particular, I. pacificus was the only species found attached to all of the infected horses. Exposure to bites of ticks in the genus Ixodes poses previously unrecognized and serious health risks to humans and animals.

Transmission and passage in horses of the agent of human granulocytic ehrlichiosis.

The human granulocytotropic ehrlichia and Ehrlichia equi produce similar diseases in their respective host species (humans, horses). Currently, the phylogenetic and biologic relationships of these 2 uncultured pathogens remain unclear. Previous studies have revealed nucleotide sequence similarity approaching identity at the level of the 16S ribosomal RNA gene. To investigate the biologic similarities of these 2 ehrlichiae, the susceptibility of horses to the human agent was tested by intravenous inoculation of infected human blood. The results demonstrate that the human granulocytotropic ehrlichia produces a disease in the horse indistinguishable from that caused by E. equi, providing biologic evidence that these 2 organisms are highly related and potentially conspecific. It is possible that cases of human illness now attributed to human granulocytotropic ehrlichia may in fact be caused by 1 or more strains of an ehrlichia known chiefly as an equine pathogen.

Vaccination against Lyme disease caused by diverse Borrelia burgdorferi.

Diversity and mutations in the genes for outer surface proteins (Osps) A and B of Borrelia burgdorferi sensu lato (B. burgdorferi), the spirochetal agent of Lyme disease, suggests that a monovalent OspA or OspB vaccine may not provide protection against antigenically variable naturally occurring B. burgdorferi. We now show that OspA or OspB immunizations protect mice from tick-borne infection with heterogeneous B. burgdorferi from different geographic regions. This result is in distinct contrast to in vitro killing analyses and in vivo protection studies using syringe injections of B. burgdorferi as the challenge inoculum. Evaluations of vaccine efficacy against Lyme disease and other vector-borne infections should use the natural mode of transmission and not be predicated on classification systems or assays that do not rely upon the vector to transmit infection.

Host association and the capacity of sand flies as vectors of lizard malaria in Panama.

In this paper the capacity of sand flies (Lutzomyia) as vectors of parasites that cause malaria in anoles (Anolis limifrons) in the Zona de Canal, Panama was investigated. Inhabiting all study plots, often in local abundance, L. trinidadensis emerged as the principal candidate sand fly vector; the results of surveys did not suggest a likely mosquito vector. Although L. trinidadensis and infected anoles co-inhabited all plots, their abundances seemed unrelated. No evidence that sand flies parasitized anoles was uncovered. As anole activity patterns in daylight reciprocate with those of sand flies and at night anoles seem to avoid locations that sand flies frequent, anoles may evade sand fly bites altogether. Further, these sand flies occurred in close numerical and ecological association with Thecadactylus rapicauda, a reclusive moist forest gecko, often parasitizing these hosts in large numbers. Thus, sand flies lack capacity as vectors of malaria-causing parasites in central Panamanian anoles.

Motility of Lyme disease spirochetes in fluids as viscous as the extracellular matrix.

When properties of extracellular fluids that might regulate the ability of the Lyme disease spirochete to locomote were investigated, the rate of progression correlated with viscoelasticity. Such spirochetes flexed and rotated but did not progress in relatively nonviscous fluids and migrated increasingly rapidly as the viscous characteristics of the medium increased. The viscoelastic properties of various kinds of hyaluronic acid resembled those of a methylcellulose standard. The maximum velocity that spirochetes achieved in such solutions related directly to viscoelasticity rather than to chemical composition. Spirochetes remained motile during 3 h of observation despite 100-fold dilution of the standard nutrient medium. The immobility of Lyme disease spirochetes in media less viscous in character than fixed tissue suggests dissemination via the intercellular ground substance of skin.

A simple electronic timer for animal-baited intermittent suction insect traps.

A versatile integrated circuit timer for intermittent suction, animal-baited insect traps is described. Various attributes of this circuit include continuously and independently variable on and off periods, extremely low power consumption, accurate timing range from microseconds through hours, operation on 5-17 VDC, and miniaturization.