Regulatory processes for approval of the contained-field testing of genetically modified <i>A. aegypti</i> in Rio Florido, Tapachula, Chiapas, Mexico.

<p>Regulatory processes for approval of the contained-field testing of genetically modified <i>A. aegypti</i> in Rio Florido, Tapachula, Chiapas, Mexico.</p

Schematic representation of the regulatory pathway for research involving genetically-modified mosquitoes.

<p>Activities of the Project principal investigator and collaborating institutions (oversight collaborating institutions) are coordinated in Mexico by the principal investigator at the Centro Regional de Investigación en Salud Pública (PI/Project Crisp). CRISP also is the regional center of the researchers conducting the work at the field site. The director general of the Instituto Nacional de Salud Publica/National Institute of Public Health (DG/INSP) is the titular head of the collaborating investigators in Mexico. The oversight committee reviews ethical, biosafety, and scientific practices used in the project and makes recommendations to the DG/INSP regarding procedures or adherence of the project participants or internal commissions to appropriate standards. The Bioethics, Research and Biosecurity Commissions are in-house INSP regulatory bodies. CIBIOGEM (Comisión Intersectorial de Bioseguridad de los Organismos Geneticamente Modificados [Intersector Commission for Biosafety of Genetically Modified Organisms]) is the federal-level commission that formulates and coordinates policies in matters related to the biosafety of genetically modified organisms. CIBIOGEM comprises the heads of SEMARNAT (Secretaria del Medio Ambiente y Recursos Naturales [Environmental and Natural Resources Secretary]), SAGARPA (Secretaria de Agricultura, Ganaderia, Desarrollo Rural, Pesca y Alimentación [Agriculture, Livestock, Rural Development, Fisheries and Food Security Secretary]), Cofepris (Comisión Federal para la Protección contra Riesgos Sanitarios [Federal Comision for the Protection against Sanitary risk])/SS (Secretaría de Salud [Secretary of Health]), and a number of others listed in the text. SEMARNAT is the authority to which the Notification (<i>Aviso</i>) was submitted regarding the importation and contained research on genetically modified organisms and that which granted a permit for the collection of native fauna to monitor transgenes. The red arrows show the permit ratification levels and information flow. State-level Sanitary Regulation (IMSS, Instituto Mexicano de Seguro Social [Mexican Social Security Institute]; ISECH, Instituto de Salud del Estado de Chiapas [Chiapas State Health Institute]) monitors and enforces health regulations at this level and ratifies SEMARNAT's permit for genetically modified organisms. County-level Health Committee ratifies ISECH/Cofepris approval for use of genetically modified organisms. Agrarian Reform grants changes in land classification and permission to purchase land and this informs the regional authorities (orange arrow and lines). The Ejido Assembly gives local permission to sell land to INSP and INDAABIN (Instituto de Administración y Avalúos de Bienes Nacionales [Appraisal and Administration of National Property Institute]) grants permission for property purchase by INSP. Catastro (land registry) gives land titles and obras publicas (public works) grants construction permits.</p

Mexican federal regulatory agencies and their laws and regulations relevant to the contained-field trial of genetically modified <i>A. aegypti</i>.

<p>Mexican federal regulatory agencies and their laws and regulations relevant to the contained-field trial of genetically modified <i>A. aegypti</i>.</p

Adult males sampled weekly with BG Sentinel Traps.

<p>Each week, adult sampling was performed the day before the weekly release of OX3604C males. Significantly higher numbers of males were collected in treatment cages with respect to control cages (<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002001#pntd.0002001.s010" target="_blank">Table S3</a>) starting from Week 2 post-release (PR), indicating that transgenic males were present in large numbers over time in release cages.</p

Egg production in treatment and control cages.

<p>Weekly egg production is shown for each control and treatment cage. Production numbers were stable in all cages by week 9 after population establishment. After OX3604C male release was initiated (vertical dashed line) in the treatment cages (week 16; week 0 PR, top time axis), egg production in the control cages continued to be stable and declined slightly in the treatment cages.</p

Progeny genotypes in treatment cages.

<p>A random sample of eggs from each treatment cage collected weekly was hatched and screened for the DsRed2 marker starting from Week 0 post-release. The number of screened larvae corresponded to 10% of the eggs produced weekly per cage or a minimum of two hundred, when available.</p

Extinction time estimated per each treatment cage assuming the estimated fitness costs from Table 1.

1<p>Minimum extinction time.</p>2<p>Maximum extinction time.</p>3<p>Mean extinction time.</p>4<p>Standard Deviation.</p>5<p>Probability of observing extinction between weeks 18–28 PR, obtained from the outcomes of 1000 simulated experiments.</p

Summary of mating competitiveness experiments.

1<p>In experiment 6 the cage number is specified.</p>2<p>At the field site location, small cages were placed in the field laboratory.</p>3<p>Number of mixed batches with the hypothesis of 1∶1 ratio of OX3604C∶GDLS2 larvae not rejected by Chi Square Test. For statistical analysis, this number was included in the number of OX3604C matings because we assumed they came from matings with heterozygous males and not from occasional double matings.</p>(*)<p> = statistically significant results.</p

OX3604C fitness cost estimates during the field cage trial per treatment cage.

1<p>Sum of square errors associated with each estimate.</p>2<p>Standard deviation.</p

ANCOVA results for egg production dynamics in paired cages A, B, C, D, and E.

<p>ANCOVA results for egg production dynamics in paired cages A, B, C, D, and E.</p