Publication: Identificación molecular de muestras de flora y fauna silvestre por medio de la técnica Barcoding, como nueva pericia en el Departamento de Ciencias Forenses de Costa Rica para combatir el tráfico ilegal
Authors
Oliva Barboza, Rossana
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Escuela Internacional de Doctorado
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Galián Albaladejo, José ; Arnaldos Sanabria, María Isabel
Publisher
Universidad de Murcia
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DOI
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info:eu-repo/semantics/doctoralThesis
Description
Abstract
El tráfico ilegal de fauna y flora silvestre es una de las amenazas más graves contra la biodiversidad y constituye la cuarta actividad ilícita más lucrativa del mundo, tras narcotráfico, falsificación y trata de personas. Este comercio global, multimillonario y complejo trasciende lo ambiental para impactar en lo legal, económico y social. Provoca disminución de poblaciones vulnerables, desestabilización de ecosistemas, pérdida de ingresos estatales y afecta comunidades dependientes del manejo sostenible de recursos. Enfrentarlo exige respuestas integrales, donde la ciencia forense aporta herramientas clave para identificar especies, vincular evidencias y sustentar procesos judiciales. Costa Rica, que alberga cerca del 6,5% de la biodiversidad mundial con más de 120.000 especies registradas, enfrenta creciente presión por delitos ambientales. Su riqueza natural y posición geográfica la convierten en punto crítico de tráfico. Entre 2014 y 2018 se registraron más de 2.500 decomisos, y entre 2012 y 2017 casi 10.000 denuncias judiciales por infracciones ambientales. Sin embargo, desde 2015 el presupuesto estatal para protección de biodiversidad es apenas un 0,1% del PIB, lo que limita la capacidad institucional, incluido el Organismo de Investigación Judicial (OIJ). En este escenario, el Departamento de Ciencias Forenses (DCF) del OIJ cumple un papel central. Su Sección de Biología Forense realiza pericias de identificación morfológica de restos zoológicos y botánicos, pero enfrenta limitaciones: depende de claves dicotómicas que exigen ejemplares completos y especialistas escasos, lo que obliga a consultar expertos externos que no siempre cumplen criterios forenses como la cadena de custodia. Además, fragmentos incompletos, estados inmaduros o especies similares dificultan identificaciones. Estas carencias pueden derivar en impunidad, pues no siempre es posible determinar si un espécimen pertenece a categorías protegidas por CITES o la UICN. Frente a estas dificultades se propone aplicar DNA Barcoding, técnica que analiza regiones estandarizadas de ADN (COI en animales, rbcL y matK en plantas) para diferenciar especies mediante comparación en bases de datos como GenBank o BOLD Systems. Desde su introducción en 2003, el Barcoding ha demostrado eficacia en taxonomía, conservación y ciencias forenses, destacando por objetividad, reproducibilidad y valor judicial. La investigación evaluó la viabilidad del Barcoding como pericia forense en Costa Rica, organizada en tres capítulos: plantas, vertebrados e insectos. Se analizaron métodos de conservación, extracción y marcadores genéticos. En plantas, se identificaron condiciones óptimas para preservar ADN y se aplicó con éxito en casos reales de botánica forense. En vertebrados, los genes COI y citocromo b superaron limitaciones de restos degradados y validaron especies de mamíferos, aves y peces en casos de cacería y comercio ilegal. En insectos se estudiaron 40 especies de dípteros y coleópteros. El Mini-COI fue muy eficaz en dípteros incluso con fragmentos degradados, mientras que en coleópteros resultaron útiles 16S y ND5, especialmente en géneros de valor comercial y conservación como Chiasognathus y Ceroglossus. Un aporte importante fue la aplicación directa de esta técnica en casos reales del DCF: autopsias que requerían determinar el intervalo post mortem mediante larvas de dípteros y decomisos de insectos asociados a redes de tráfico. En todos los escenarios, el Barcoding complementó la morfología tradicional y reforzó la solidez de los dictámenes técnicos. La investigación concluye que la identificación morfológica sigue siendo fundamental, pero debe complementarse con enfoques moleculares. La eficiencia del Barcoding depende de cada grupo biológico y marcador utilizado, por lo que requiere metodologías adaptadas. Su implementación en Costa Rica es viable y necesaria: fortalecería al DCF, apoyaría la lucha contra el tráfico ilegal y potenciaría la protección de la biodiversidad. Además, permitiría crear una base de datos nacional de referencia, consolidando al país como líder regional en la aplicación de genética molecular a las ciencias forenses ambientales.
Wildlife trafficking is among the most serious threats to biodiversity and the world’s fourth most lucrative illicit activity, after drug trafficking, counterfeiting, and human trafficking. This global, multimillion-dollar trade transcends environmental impacts to affect legal, economic, and social dimensions. It causes declines of vulnerable populations, destabilization of ecosystems, loss of state revenues, and harms communities dependent on sustainable resource use. Addressing it requires integrated responses, with forensic science providing key tools to identify species, link evidence, and support judicial processes. Costa Rica, harboring about 6.5% of global biodiversity with more than 120,000 recorded species, faces growing pressure from environmental crimes. Its natural wealth and geographic position make it a critical hub. Between 2014 and 2018, over 2,500 seizures were registered, and between 2012 and 2017 nearly 10,000 judicial complaints for environmental offenses. Yet since 2015, state funding for biodiversity protection has been just 0.1% of GDP, limiting institutional capacity, including the Judicial Investigation Agency (OIJ). In this scenario, the OIJ’s Department of Forensic Sciences (DCF) plays a central role. Its Biology Section conducts morphological identifications of zoological and botanical remains but faces limitations: reliance on dichotomous keys requiring complete specimens and scarce specialists, often forcing consultations with external experts who may not follow forensic standards such as chain of custody. Incomplete fragments, immature stages, or morphologically similar species further hinder identifications. These gaps may result in impunity, since it is not always possible to determine whether a specimen belongs to categories protected by CITES or IUCN. To address these challenges, DNA Barcoding is proposed, analyzing standardized DNA regions (COI in animals, rbcL and matK in plants) to differentiate species through databases like GenBank or BOLD Systems. Since its introduction in 2003, Barcoding has proven effective in taxonomy, conservation, and forensic science, valued for objectivity, reproducibility, and judicial credibility. This study assessed the feasibility of Barcoding as a forensic tool in Costa Rica, structured in three chapters: plants, vertebrates, and insects. Methods of preservation, extraction, and genetic markers were evaluated. In plants, optimal conditions for DNA preservation were identified and applied successfully in real forensic cases. In vertebrates, COI and cytochrome b overcame degraded sample issues and validated species of mammals, birds, and fish in hunting and illegal trade cases. In insects, 40 dipteran and coleopteran species were studied. Mini-COI proved highly effective in dipterans even with degraded fragments, while 16S and ND5 were useful in coleopterans, particularly in commercially and conservation-relevant genera such as Chiasognathus and Ceroglossus. An important contribution was the direct application of this technique in real DCF cases: autopsies requiring postmortem interval estimation with dipteran larvae, and insect seizures linked to trafficking networks. In all scenarios, Barcoding complemented traditional morphology and reinforced the reliability of technical reports. The study concludes that morphological identification remains essential but must be complemented with molecular approaches. Barcoding’s efficiency depends on each biological group and marker, requiring adapted methodologies. Its implementation in Costa Rica is viable and necessary: it would strengthen the DCF, support the fight against wildlife trafficking, and enhance biodiversity protection. It would also enable a national reference database, consolidating the country as a regional leader in applying molecular genetics to environmental forensic science.
Wildlife trafficking is among the most serious threats to biodiversity and the world’s fourth most lucrative illicit activity, after drug trafficking, counterfeiting, and human trafficking. This global, multimillion-dollar trade transcends environmental impacts to affect legal, economic, and social dimensions. It causes declines of vulnerable populations, destabilization of ecosystems, loss of state revenues, and harms communities dependent on sustainable resource use. Addressing it requires integrated responses, with forensic science providing key tools to identify species, link evidence, and support judicial processes. Costa Rica, harboring about 6.5% of global biodiversity with more than 120,000 recorded species, faces growing pressure from environmental crimes. Its natural wealth and geographic position make it a critical hub. Between 2014 and 2018, over 2,500 seizures were registered, and between 2012 and 2017 nearly 10,000 judicial complaints for environmental offenses. Yet since 2015, state funding for biodiversity protection has been just 0.1% of GDP, limiting institutional capacity, including the Judicial Investigation Agency (OIJ). In this scenario, the OIJ’s Department of Forensic Sciences (DCF) plays a central role. Its Biology Section conducts morphological identifications of zoological and botanical remains but faces limitations: reliance on dichotomous keys requiring complete specimens and scarce specialists, often forcing consultations with external experts who may not follow forensic standards such as chain of custody. Incomplete fragments, immature stages, or morphologically similar species further hinder identifications. These gaps may result in impunity, since it is not always possible to determine whether a specimen belongs to categories protected by CITES or IUCN. To address these challenges, DNA Barcoding is proposed, analyzing standardized DNA regions (COI in animals, rbcL and matK in plants) to differentiate species through databases like GenBank or BOLD Systems. Since its introduction in 2003, Barcoding has proven effective in taxonomy, conservation, and forensic science, valued for objectivity, reproducibility, and judicial credibility. This study assessed the feasibility of Barcoding as a forensic tool in Costa Rica, structured in three chapters: plants, vertebrates, and insects. Methods of preservation, extraction, and genetic markers were evaluated. In plants, optimal conditions for DNA preservation were identified and applied successfully in real forensic cases. In vertebrates, COI and cytochrome b overcame degraded sample issues and validated species of mammals, birds, and fish in hunting and illegal trade cases. In insects, 40 dipteran and coleopteran species were studied. Mini-COI proved highly effective in dipterans even with degraded fragments, while 16S and ND5 were useful in coleopterans, particularly in commercially and conservation-relevant genera such as Chiasognathus and Ceroglossus. An important contribution was the direct application of this technique in real DCF cases: autopsies requiring postmortem interval estimation with dipteran larvae, and insect seizures linked to trafficking networks. In all scenarios, Barcoding complemented traditional morphology and reinforced the reliability of technical reports. The study concludes that morphological identification remains essential but must be complemented with molecular approaches. Barcoding’s efficiency depends on each biological group and marker, requiring adapted methodologies. Its implementation in Costa Rica is viable and necessary: it would strengthen the DCF, support the fight against wildlife trafficking, and enhance biodiversity protection. It would also enable a national reference database, consolidating the country as a regional leader in applying molecular genetics to environmental forensic science.
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