In vitro establishment and propagation of a Brazilian strain ofAnaplasma marginale with appendage in IDE8 (Ixodes scapularis) cells

Abstract

A Brazilian isolate of Anaplasma marginale with appendage was successfully established and maintained in vitro in a tick cell line (IDE8). Infection was confirmed by optical and transmission electron microscopy. In addition, primers MSP1aNF2 and MSP1aNR2 amplified products from DNA extracted from infected IDE8 cells. Comparisons with partial sequences of the msp1α gene and the complete genome of A. marginale confirmed that the sequences of amplified fragments were from the A. marginale genome. This is the first establishment of a Brazilian A. marginale isolate in tick cells, representing a new system for biological and molecular studies and also a new source of material for diagnosis and development of vaccines.

Characterization of two strains of Anaplasma marginale isolated from cattle in Rio de Janeiro, Brazil, after propagation in tick cell culture

“…the organisms and thus slowing down culture development. Otherwise, the development of the colonies in the tick cells was similar to that already described (Bastos et al, 2009;Blouin et al, 2000). The sequence of the msp1˛ gene of A. marginale from both cattle remained the same after passage in IDE8 tick cell line.…”

“…First colonies were detected 60 days after culture initiation. The time until first detection, however, was considerably longer than reported in other studies (Bastos et al, 2009;Blouin et al, 2000;Munderloh et al, 1996) where periods between 8 and 34 days were observed. The reasons for this could have been manifold.…”

“…The first continuous in vitro culture system for the pathogen A. marginale was established in the Ixodes scapularis cell line IDE8 from infected bovine erythrocytes (Munderloh et al, 1996). Subsequently, this system has been used for the isolation and propagation of several isolates of A. marginale (Bastos et al, 2009;Blouin et al, 2002).…”

In vitro cultivation of Anaplasma marginale and A. phagocytophilum in tick cell lines: a review

“…marginale cultures can be initiated from blood obtained from infected calves during ascending rickettsemia (BLOUIN et al, 2000;BASTOS et al, 2009). Blood samples are collected with anticoagulant, and usually the blood cells are washed and subsequently cryopreserved in liquid nitrogen using DMSO as cryoprotectant.…”

Evaluating the effectiveness of an inactivated vaccine from Anaplasma marginale derived from tick cell culture

“…Culturing of Brazilian strains of A. marginale in IDE8 cells was described previously (BASTOS et al, 2009). Briefly, monolayers of IDE8 cells were inoculated with blood containing UFMG1 and UFMG2 isolates from A. marginale that had been cryopreserved in liquid nitrogen.…”