Susceptibility of mice (Mus musculus) to repeated infestation with Amblyomma americanum (Acari: Ixodidae) ticks.

Laboratory mice, Mus musculus (L.), BALB/c strain, were assessed for their ability to develop resistance to repeated infestation by Amblyomma americanum (L.) ticks. Mice were infested five consecutive times with A. americanum nymphs. No decrease in tick viability was seen after five infestations, suggesting that BALB/c mice do not develop immune-mediated resistance to A. americanum. In contrast, tick viability was significantly reduced in the second infestation of a New Zealand White rabbit, a laboratory animal known to develop resistance to A. americanum.

Reinfection with Anaplasma phagocytophilum in BALB/c mice and cross-protection between two sympatric isolates.

Infection with Anaplasma phagocytophilum in white-footed mice results in partial protection against reinfection with the same agent. However, humans and domestic animals may be sequentially exposed to different isolates of the agent circulating in the same or adjacent foci. We investigated whether immune response to a tick-borne infection with A. phagocytophilum provides protection against homologous and heterologous challenges. BALB/c mice were infected with one of the two sympatric isolates of A. phagocytophilum via tick bite and challenged 16 weeks later by Ixodes scapularis nymphs infected with either the same or the alternative isolate. As controls, groups of infected mice were challenged by uninfected ticks to confirm an absence of reactivation of the original infection or groups of naive mice were fed upon by ticks from cohorts used for an infectious challenge. Xenodiagnostic I. scapularis larvae were fed upon each mouse at 14 and 21 days postchallenge (PCH) and tested for the presence of A. phagocytophilum as freshly molted nymphs. Blood samples for quantitative PCR were collected at 7, 14, 21, and 70 days PCH. Serum samples were collected weekly to monitor development of immune response. The proportion of infected animals, levels of bacteremia, and the prevalence of infection in xenodiagnostic ticks were higher in groups of control mice exposed to A. phagocytophilum for the first time than in mice reinfected with either homologous or heterologous isolates. The presence of antibodies against A. phagocytophilum did not protect mice from a challenge with either homologous or heterologous isolates, however the ensuing reinfection was significantly milder and of a shorter duration than the first infection with either isolate.

Effects of Anaplasma phagocytophilum infection on the molting success of Ixodes scapularis (Acari: Ixodidae) larvae.

We assessed the effects of sympatric (occupying the same or overlapping geographic areas) and allopatric (occurring in separate geographic areas) isolates of Anaplasma phagocytophilum on the survival of Ixodes scapularis Say larvae that were derived from ticks collected in Bridgeport, CT. Seven isolates of A. phagocytophilum, originating from different geographic regions of the United States, were tested: four isolates from the northeast (Bridgeport, Dawson, Gaillard, and NY-8), two from the Midwest (Webster and Sp-Is), and one from California (MRK). BALB/c mice were infected with each of the seven isolates via exposure to infected I. scapularis nymphs, whereas uninfected nymphs fed upon control mice. Both infected and control mice were infested with uninfected larvae at 1, 2, 3, 4, 6, and 9 wk after nymphal infestation. The molting success in cohorts of infected and uninfected ticks was calculated as the percentage of larvae successfully molting into nymphal stage, and the prevalence of infection in molted nymphs was determined by polymerase chain reaction. In ticks that became infected with the Bridgeport or Sp-Is isolates, the molting success decreased with an increase in the prevalence of infection. Ticks that fed upon mice infected with six allopatric isolates (Dawson, Gaillard, NY-8, Sp-Is, Webster, and MRK) showed significantly lower levels of survival than those fed upon control mice, regardless of the prevalence of infection, whereas in ticks fed upon mice infected with a sympatric isolate (Bridgeport), the overall molting success was similar to the control. Thus, some but not all of the A. phagocytophilum isolates have adverse effects on ticks. Ticks exposed to harmful isolates may experience higher levels of bacterial metabolism, and/or reduced quality of their blood meal, thereby reducing their survival. Noted differences between isolates may be due to the origin of a particular isolate and/or the degree of coadaptation between the pathogen and its vector on the population level.

Acquisition of different isolates of Anaplasma phagocytophilum by Ixodes scapularis from a model animal.

The prevalence of etiologic agents in ticks reflects the intensity of their transmission in natural cycles and is an important measure of their potential to cause human disease. The distribution of Anaplasma phagocytophilum within the range of its primary vectors is patchy. Even nearby sites differ dramatically in the prevalence of Anaplasma in questing ticks. We hypothesized that this irregular distribution may be due in part to variations in acquisition rates of different isolates of A. phagocytophilum by I. scapularis ticks from infected animals. BALB/c mice were infected with seven isolates of A. phagocytophilum from different geographical regions: four isolates from the Northeastern United States (Bridgeport, Dawson, Gaillard, NY-8), two from the Midwest (Webster, Sp-Is), and one from California (MRK). Larval ticks were placed on infected mice for 16 consecutive weeks and allowed to feed to repletion. The prevalence of infection in the freshly molted nymphs was then determined by PCR. The proportion of ticks that became infected with either isolate fluctuated over the duration of infection. Mice harboring the isolate Sp-Is were most infectious for ticks at 3 weeks postinfection. Mice infected with the other six isolates exhibited several peaks of infectivity. Timing and relative heights of these peaks differed between isolates. Geographical proximity of the studied isolates did not predetermine their similarity, and isolates originating from the same region differed more in their ability to infect ticks than isolates from different regions. However, it appears unlikely that described differences in agent acquisition by ticks alone are sufficient to account for the irregular distribution of A. phagocytophilum in vector populations.