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Background  Paratransgenesis is an approach to reducing arthropod vector competence using genetically modified symbionts. When applied to control of Chagas disease, the symbiont bacterium  Rhodococcus rhodnii , resident in the gut lumen of the triatomine vector  Rhodnius prolixus  (Hemiptera: Reduviidae), is transformed to export cecropin A, an insect immune peptide. Cecropin A is active against  Trypanosoma cruzi , the causative agent of Chagas disease. While proof of concept has been achieved in laboratory studies, a rigorous and comprehensive risk assessment is required prior to consideration of field release. An important part of this assessment involves estimating probability of transgene horizontal transfer to environmental organisms (HGT). This article presents a two-part risk assessment methodology: a theoretical model predicting HGT in the gut of  R. prolixus  from the genetically transformed symbiont  R. rhodnii  to a closely related non-target bacterium,  Gordona rubropertinctus , in the absence of selection pressure, and a series of laboratory trials designed to test the model.    Results  The model predicted an HGT frequency of less than 1.14 × 10 -16  per 100,000 generations at the 99% certainty level. The model was iterated twenty times, with the mean of the ten highest outputs evaluated at the 99% certainty level. Laboratory trials indicated no horizontal gene transfer, supporting the conclusions of the model.    Conclusions  The model treats HGT as a composite event, the probability of which is determined by the joint probability of three independent events: gene transfer through the modalities of transformation, transduction, and conjugation. Genes are represented in matrices and Monte Carlo method and Markov chain analysis are used to simulate and evaluate environmental conditions. The model is intended as a risk assessment instrument and predicts HGT frequency of less than 1.14 × 10 -16  per 100,000 generations. With laboratory studies that support the predictions of this model, it may be possible to argue that HGT is a negligible consideration in risk assessment of genetically modified  R. rhodnii  released for control of Chagas disease.

Modeling horizontal gene transfer (HGT) in the gut of the Chagas disease vector Rhodnius prolixus