The decomposition of a body, and the associated gaseous and liquid discharges emanating from it, attract gravid female blow flies which lay their eggs in or on the body. After the eggs have hatched, the emerging larvae start feeding on the body. As decomposition progresses, the blow fly larvae often migrate away, typically in a random manner in search of favourable conditions for pupation. In this paper, we report on a rarely described phenomenon of unidirectional mass migration of blow fly larvae and postulate on the factors which may drive this process. A decomposition trial utilising a 60-kg pig carcass, deployed in the summer months in Table Mountain National Park, Cape Town, South Africa, was conducted in 2022. On the fifth day of the trial, simultaneous unidirectional mass dispersal of blow fly larvae was observed. The larvae moved downhill in a southeasterly direction, following the flow of decomposition fluids oozing out from the pig carcass. The 'larval migration stream' had a length of approximately 1.5 m with a width of 40 cm, tapering to 17 cm at the terminal point. The larval migration stream consisted of the larvae of Chrysomya albiceps and Chrysomya chloropyga. This study demonstrates the importance of understanding the timing and pattern of dispersal of post-feeding blow fly larvae in each geographical region. This is crucial as the minimum post-mortem interval can be miscalculated if older immature insects dispersing from the corpse are not considered and collected during crime scene investigations. SIGNIFICANCE: • Understanding the timing and pattern of dispersal of blow fly larvae from decomposing vertebrate remains in each geographical region is important in time-since-death estimations when using entomological evidence. • In the summer season of the Western Cape Province, Chrysomya albiceps and Chrysomya chloropyga are among the blow fly species of forensic importance as they deposit their eggs on vertebrate remains. • When a decomposing body is found within the Table Mountain region, we suggest that crime scene investigators should examine the downhill area of the scene for larval/pupal entomological evidence that can assist in time and season of death estimations.

Resumen La entomotoxicología forense, que analiza insectos necrófagos para detectar sustancias tóxicas, es crucial en la determinación de causas de muerte y la estimación del intervalo post-mortem. Este estudio se centró en Peckia intermutans (Walker) (Diptera: Sarcophagidae) como marcador para la detección de metanol, un alcohol potencialmente letal presente como adulterante en bebidas alcohólicas debido a su bajo costo y parecido con el etanol. Se desarrolló un método de cromatografía de gases-espectrometría de masas (HS-GC-MS) para detectar metanol y se estudió su efecto sobre el tiempo de desarrollo, supervivencia y morfología de las moscas de la carne. Las larvas se criaron en sustratos de hígado enriquecidos con metanol (300, 500 y 1000 mg/kg). Los resultados mostraron: el método desarrollado detectó metanol en todos los estadios; el metanol afectó el tiempo de desarrollo de las moscas; la supervivencia aumentó con una mayor concentración de metanol; y los cambios morfológicos fueron evidentes en el peso. Este estudio ofrece valiosa información sobre la investigación entomotoxicológica del Ecuador, por lo que se debe considerar los cambios morfológicos de P. intermutans para evitar estimaciones inexactas de los intervalos post-mortem.

Abstract Forensic entomotoxicology, which analyzes necrophagous insects to detect toxic substances, is crucial in determining causes of death and estimating the post-mortem interval. This study focused on Peckia intermutans (Walker) (Diptera: Sarcophagidae) as a marker for detecting methanol, a potentially lethal alcohol often added as an adulterant in alcoholic beverages due to its low cost and similarity to ethanol. A gas chromatography-mass spectrometry (HS-GC-MS) method was developed to detect methanol, and its effects on the development time, survival, and morphology of flesh flies were investigated. Larvae were reared on liver substrates enriched with methanol (300, 500, and 1000 mg/kg). Results showed: the developed method detected methanol in all instars; methanol affected the developmental time of the flies; survival increased with higher methanol concentration; and morphological changes were evident in weight. This study provides valuable information on entomotoxicological research in Ecuador, emphasizing the need to consider morphological changes in P. intermutans to avoid inaccurate estimations of post-mortem intervals.

Abstract Chrysomya albiceps (Calliphoridae) is among the earliest successional fauna on human and animal cadavers. Some immature Calliphoridae can be useful for determination of post-mortem interval. Toxins, particularly pesticides, can affect the rate of insect growth. Aluminum phosphide (AlP) is an affordable insecticide that has not been adequately entomotoxicologically evaluated. So, the impact of AlP on the developmental rate of different stages of C. albiceps was investigated. Larvae of C. albiceps were reared on the rabbit carcasses containing AlP as a treated group, and distilled water as a control group. The substances were administered by a gastric tube. The duration needed for development of C. albiceps stages was documented. Body length, width and weight of larvae were measured after 24, 48, 72 and 96 h from egg hatching. The duration of development increased significantly in the treated group compared to the control group. Larvae body measurements were significantly smaller in the treated group than in the control group. Therefore, it was demonstrated that AlP significantly influences the size of C. albiceps larvae and extends their development. During forensic application, interpretation of C. albiceps data should be used with caution when aluminum phosphide may be the cause of death. (Calliphoridae cadavers postmortem post mortem interval Toxins pesticides growth (AlP evaluated So C investigated tube documented length 24 48 7 9 hatching Therefore application death 2 4

Resumo Chrysomya albiceps (Calliphoridae) está entre as primeiras espécies a colonizar cadáveres humanos e animais. Alguns Calliphoridae imaturos podem ser úteis para a determinação do intervalo “post-mortem”. Toxinas, particularmente pesticidas, podem afetar a taxa de crescimento de insetos. O fosfeto de alumínio (AlP) é um inseticida acessível que não foi adequadamente avaliado entomotoxicologicamente. Assim, o impacto do AlP na taxa de desenvolvimento de diferentes estágios de C. albiceps foi investigado. Larvas de C. albiceps foram criadas em carcaças de coelho, contendo AlP como grupo tratado e água destilada como grupo controle. As substâncias foram administradas por sonda gástrica. A duração necessária para o desenvolvimento dos estágios de C. albiceps foi documentada. O comprimento, largura e peso do corpo das larvas foram medidos após 24, 48, 72 e 96 horas da eclosão dos ovos. A duração do desenvolvimento foi significativamente aumentada no grupo tratado, em comparação com o grupo controle. As medidas corporais das larvas foram significativamente menores no grupo tratado do que no grupo controle. Portanto, foi demonstrado que o AlP influencia significativamente o tamanho das larvas de C. albiceps e prolonga seu desenvolvimento. Durante a aplicação forense, a interpretação dos dados de C. albiceps deve ser usada com cautela, quando o fosfeto de alumínio puder ser a causa da morte. (Calliphoridae animais postmortem. postmortem post mortem . “post-mortem” Toxinas pesticidas insetos (AlP entomotoxicologicamente Assim C investigado coelho controle gástrica documentada comprimento 24 48 7 9 ovos Portanto forense cautela morte “post-mortem 2 4

Abstract Objective: To study the impact of age and the interval between disease diagnosis and death on the organotropism of SARS-CoV-2. Method: Patients underwent post-mortem biopsies from lungs, Waldeyer ring, heart, liver, kidneys and bone marrow between 2020–2021. SARS-CoV-2 organotropism was mapped by using molecular RT-PCR analyses for SARS-CoV2 targeting the Envelope gene (E), the RNA Polymerase Gene (RdRp), and the Nucleocapsid gene (N). Statistical and linear regression analysis was performed to study the impact of age and illness duration in SARS-CoV-2 organotropism. Results: We performed 158 postmortem biopsies in 21 patients, with a mean age of 76 years old. The mean interval between the diagnosis of the infection to the death was 23 days. The RNA of the SARS-CoV-2 was detected in 100% of lung biopsies, 76%–82% of Waldeyer’s ring biopsies, 55% of heart biopsies, 40% of kidney biopsies, 33% of liver and 25% of bone marrow biopsies. Patients who died before the day 9, presented extensive visceral dissemination of SARS-CoV-2 RNA. Most of the patients older than 80 years (90%) presented visceral dissemination of SARS-CoV-2 RNA, while among younger patients, only 3/11 patients presented visceral dissemination of the virus. The relationship between “age” and “illness duration” and multitropism of the virus was statistically significant (p< 0.001). Conclusion: Disease interval and age were factors that were significantly associated with RT-PCR positive results in multiple organs. Critical COVID-19 patients have multiorganic viral dissemination in early stages. In the critical older patients, multiorganic viral dissemination is the rule. Level of evidence: 4. Case Series. Objective SARSCoV2. SARSCoV2 SARSCoV SARS CoV 2. 2 Method post mortem lungs 20202021 2020 2021 2020–2021 SARS-CoV- RTPCR RT PCR CoV2 SARS-CoV E, E , (E) RdRp, RdRp (RdRp) N. N . (N) Results 15 7 old days 100 7682 82 76%–82 Waldeyers s 55 40 33 25 9 8 90% 90 (90% 311 3 11 3/1 “age p< p (p 0.001. 0001 0.001 0 001 0.001) Conclusion organs COVID19 COVID 19 COVID-1 stages rule evidence 4 Series 2020202 202 2020–202 (E (RdRp (N 1 10 768 76%–8 5 (90 31 3/ 000 0.00 00 COVID1 COVID- 202020 20 2020–20 76%– (9 0.0 20202 2020–2 76% ( 0. 2020–

Resumen Calliphora vicina Robineau-Desvoidy es una especie colonizadora primaria en cuerpos en descomposición siendo predominante en las estaciones frías en Provincia de Buenos Aires, Argentina. Se estimó el intervalo post-mortem mínimo (PMImin) usando el crecimiento y longitud larvales como evidencia entomológica precisa del tiempo de muerte transcurrido. Se criaron especímenes a 12, 14, 16 y 18 °C para determinar las tasas de crecimiento larvales y pupales. Se registraron diferencias en los tiempos de desarrollo (43,2 días a 12 °C; 34,3 días a 14 °C; 29,1 días a 16 °C y 24,3 días a 18 °C), observándose tiempos de vida más cortos a mayor temperatura, sin alterarse los rangos de longitud larval (~1,5–18,0 mm). La temperatura también afectó la supervivencia y tamaño de los adultos emergidos; registrándose mayor porcentaje de supervivencia a 14 (29,73%) y 16 °C (38,65%) y mayor mortalidad a temperaturas extremas (19,00% a 12 °C y 12,61% a 18 °C). Los adultos criados a 12 °C fueron los más grandes, las hembras mayores que los machos con una relación inversa entre tamaño y supervivencia. Estos resultados permiten estimar el PMI basado en datos de poblaciones locales en lugar de consultar registros obtenidos en otras regiones.

Abstract Calliphora vicina Robineau-Desvoidy is a primary colonizing species of decomposing bodies, being predominant in the cold seasons in Buenos Aires province, Argentina. The minimum post-mortem interval (PMImin) was estimated using growth and larval length parameters as the accurate entomological evidence of the elapsed time since death. Specimens were reared at 12, 14, 16, and 18 °C, and the larval and pupal growth rates were determined. Differences in the developmental times were registered (43.2 days at 12 °C, 34.3 days at 14 °C, 29.1 days at 16 °C, and 24.3 days at 18 °C), with the lifespan shortened at higher temperatures, without changing the larval length range (~1.5–18.0 mm). Temperature also influenced the survival and size of emerged adults; the highest percentage of survival registered at 14 (29.73%) and 16 °C (38.65%), and more mortality in extreme temperatures (19.00% at 12 °C and 12.61% at 18 °C). Adults reared at 12 °C were the largest of all, females were longer than males, and body size showed an inverse relationship with survival. These results allowed the PMI estimation based on data from local populations instead of consulting papers based on other regions.

Abstract Massively parallel sequencing, also referred to as next-generation sequencing, has positively changed DNA analysis, allowing further advances in genetics. Its capability of dealing with low quantity/damaged samples makes it an interesting instrument for forensics. The main advantage of MPS is the possibility of analyzing simultaneously thousands of genetic markers, generating high-resolution data. Its detailed sequence information allowed the discovery of variations in core forensic short tandem repeat loci, as well as the identification of previous unknown polymorphisms. Furthermore, different types of markers can be sequenced in a single run, enabling the emergence of DIP-STRs, SNP-STR haplotypes, and microhaplotypes, which can be very useful in mixture deconvolution cases. In addition, the multiplex analysis of different single nucleotide polymorphisms can provide valuable information about identity, biogeographic ancestry, paternity, or phenotype. DNA methylation patterns, mitochondrial DNA, mRNA, and microRNA profiling can also be analyzed for different purposes, such as age inference, maternal lineage analysis, body-fluid identification, and monozygotic twin discrimination. MPS technology also empowers the study of metagenomics, which analyzes genetic material from a microbial community to obtain information about individual identification, post-mortem interval estimation, geolocation inference, and substrate analysis. This review aims to discuss the main applications of MPS in forensic genetics.

Abstract Caves are habitats characterized by low light or total darkness, whose fauna is largely composed of animals that adapt to dark environments, such as bats and many arthropods. Here, we present the first record of bat carcass colonization by Diptera larvae in a cave in the Neotropics. Twenty-one adult specimens of Microcerella halli (Sarcophagidae), seven males and 14 females, emerged from larvae collected in a carcass of Pteronotus gymnonotus (Mormoopidae) found in a twilight zone, about 15 m from the cave entrance. The mean time between carcass collection and adult emergence was 17.99 ± 0.44 days. As colonization by M. halli only occurs after death, we estimated the mean duration of the pupal stage - by recording daily pupation and emergence data - and attempted a preliminary estimate of the post-mortem interval, which indicated that larviposition occurred at least 24 h before carcass collection. Furthermore, we discussed the fact that M. halli females can find and colonize carcasses in low light environments, reinforcing the forensic potential of the species.

Resumen Una de las dificultades encontradas es la correcta identificación de insectos asociados a la descomposición cadavérica, por la cual ha llevado a buscar y generar nuevas herramientas en biología molecular que facilitan la determinación de especímenes para la estimación del Intervalo Post-Mortem de una forma efectiva y certera a partir de estadios inmaduros; la colecta y taxonomía morfológica de Dípteros se realizaron en primera instancia y posteriormente se utilizó el sistema de Códigos de Barras (COI Barcode) para la identificación molecular de insectos problema por medio del gen mitocondrial COI en cualquier fase del ciclo biológico. Identificando tres especies de adultos con una probabilidad de correspondencia del 100%; los especímenes: Sarconesia versicolor de la Familia Calliphoridae y Fannia sp., no fueron hallados en las bases de datos mundiales del GenBank y del Boldsystems, siendo necesario su actualización realizando patrones de sucesión cronológica de fauna cadavérica en diferentes zonas geográficas, cuya práctica se aplicaría en las investigaciones criminales.

Abstract One of the difficulties encountered is the correct identification of insects associated with cadaveric decomposition, which has led to the search and generation of new tools in molecular biology that facilitate the determination of specificities for the modification of the Post-Mortem Interval in an effective and accurate from immature stages; the collection and morphological taxonomy of Diptera were made in the first instance and then the Bar Code System (COI Barcode) was used for the molecular identification of problem insects by means of the mitochondrial COI gene in any phase of the biological cycle. Identifying three species of adults with a probability of 100% correspondence; the specimens: Sarconesia versicolor of the Calliphoridae Family and Fannia sp., were not found in the world databases of the GenBank and the Boldsystems, being necessary to update them by chronological succession patterns of cadaveric fauna in different geographical areas, whose practice would be applied in criminal investigations.

RESUMO Devido à importância médica, veterinária, ambiental e forense do califorídeo Chrysomya putoria (Diptera), foi avaliado o desempenho apresentado por estágios imaturos criados em carne de cavalo. Os espécimes foram colocados em câmaras aclimatizadas reguladas a 30 ºC, 60 ± 10% UR e 14 horas de fotofase. Os estágios larval e pupal concluíram seu desenvolvimento em 4,16 e 4,12 dias, respectivamente. As larvas pós-alimentação e pupas com 24h de idade pesaram, em média, 58,06 e 40,10mg, respectivamente. As taxas de emergência registradas foram de 65% nas condições experimentais propostas. Como os compostos orgânicos voláteis são diferentes na decomposição de um animal para outro, este trabalho e outros realizados com a mesma dieta são os mais apropriados para estimativa de intervalo post mortem envolvendo cavalos.

ABSTRACT Due to medical, veterinary, environmental and forensic importance of the calliphorid Chrysomya putoria (Diptera), the performance presented by immature stages, reared on horse meat, was evaluated. The specimens were placed in acclimatized chambers regulated at 30 ºC, 60 ± 10% RU and 14 hours of photo phase. The larval and pupal stages completed their development in 4, 16 and 4, 12 days, respectively. Post-feeding larvae and aged 24h pupae weight on average 58, 06 and 40, 10 mg, respectively. The emergence rates recorded were 65% under the experimental conditions proposed. As volatile organic compounds are different in the decomposition of one animal to another, this work and others carried out with the same diet are the most appropriate for estimating post-mortem interval involving horses.

The use of forensic entomology is well established in the northern hemisphere, but is still emerging in the southern hemisphere, where most of the current research is not explicitly undertaken in the context of forensics. In this review, we provide an update on the current status of forensic entomology research and its application in relation to estimation of post-mortem interval in various criminal investigations ranging from murder cases, cases of human neglect and the poaching of wildlife in southern Africa, among other issues. A literature search was conducted using Google Scholar, PubMed, Scopus and EBSCOhost databases. The studies reviewed were focused on arthropod diversity during different stages of carcass decomposition, effect of seasons on the abundance and diversity of carrion feeding arthropod species during carcass decomposition, and diurnal and nocturnal oviposition of forensically important insect species during carcass decomposition. It was further observed that arthropod species that established on a decomposing carcass are potentially useful in the estimation of post-mortem interval and determining clues in cases of criminal investigations. The review confirmed the paucity of research in forensic entomology, and its application in southern Africa. Future studies on the research and application of forensic entomology in various criminal investigation scenarios - such as murder cases, human neglect, and wildlife poaching in southern Africa - are therefore needed. SIGNIFICANCE: • Forensic entomology research and its application is lagging in southern Africa. • There is seasonal variation in the arthropod species used for estimation of post-mortem intervals in southern Africa. • Identification of arthropod species diversity in the region has potential for application in forensic investigations.

Resumen La identificación del género Lucilia es importante cuando se recolectan adultos o larvas en casos forenses porque el intervalo post-mortem puede ser estimado. En este trabajo se analizó morfológica y molecularmente un panel de especímenes de Lucilia (n = 42) recolectados en Costa Rica, de los cuales el 21% de las muestras corresponden a moscas criadas a partir de larvas tomadas de cadáveres humanos. Morfológicamente, los ejemplares fueron identificados como Lucilia cuprina (1) (33,3% de los ejemplares), Lucilia eximia (1) (33,3%), Lucilia purpurascens (2) (21,4%) o miembros del complejo Lucilia (11,9%). La identificación molecular, con las regiones de la citocromo oxidasa I (COI) o de la subunidad ribosomal 28S, sólo fue posible para el 43% de las muestras. La mayoría de las secuencias obtenidas con COI fueron concordantes con los análisis morfológicos, pero la región 28S carecía de suficiente resolución para identificar muestras al nivel de especies, excepto para L. cuprina. Juntos, los datos genéticos y morfológicos indican que L. cuprina y L. eximia fueron las especies más comúnmente encontradas; las moscas criadas a partir de larvas tomadas de cadáveres humanos correspondieron sólo a estas especies. Hasta donde sabemos, este trabajo es el primero en Costa Rica y en la región centroamericana en describir la entomofauna cadavérica del género Lucilia, lo cual es valioso para el desarrollo de potenciales aplicaciones forenses.

Abstract The identification of the genus Lucilia is important when adults or larvae are collected in forensic cases because the post-mortem interval might be estimated. In this work, we analyzed morphologically and molecularly a panel of Lucilia specimens (n = 42) collected in Costa Rica of which 21 % of the samples correspond to flies reared from larvae taken from human corpses. Morphologically, specimens were identified as Lucilia cuprina (1) (33.3 % of specimens), Lucilia eximia (1) (33.3 %), Lucilia purpurascens (2) (21.4 %) or members of Lucilia complex (11.9 %). Molecular identification with cytochrome oxidase I (COI) or 28S ribosomal subunit regions was possible for only 43 % of the samples. Most sequences obtained with COI were according to a morphological analysis, but the 28S region lacked sufficient resolution to identify samples to the level of species except L. cuprina. Together, genetic and morphological data indicate that L. cuprina and L. eximia were the most commonly found species; flies reared from larvae taken from human cadavers correspond to only these species. To our knowledge, this work is the first in Costa Rica and the Centro-American region to describe cadaveric entomofauna of Lucilia genus, which is valuable for the development of potential forensic applications.

Abstract The order Coleoptera is considered second highest in forensic importance in criminal cases involving violent death, because members of this order increase in richness and abundance as the decomposition process progresses. We present here the first Forensic Entomology (FE) case in Brazil where the carrion beetle, Oxelytrum discicolle (Coleoptera: Silphidae), was used to estimate the minimum post-mortem interval (m-PMI) in a forensic case. A female corpse, at an advanced stage of decomposition, was found on a deserted highway near Brasília/Brazil. The corpse was removed from the site, and insect samples were collected on and inside the corpse at the Forensic Medicine Institute of the Federal Police, following the specific FE protocols. The m-PMI was estimated considering two techniques, the pre-appearance interval (PAI) and the accumulated degree-days (ADD). The development stage of the larvae of O. discicolle allowed investigators to propose that this species was part of an earlier colonization, soon after death, with total length and prothorax width compatible with third instar larvae. Adult females of O. discicolle oviposited on the corpse at least 20 days before it was removed from the site where it was found, thus characterizing the m-PMI. This is the first case in which information on the development of the O. discicolle was used in a criminal investigation in Brazil.

Resumen Objetivos: Analizar la sucesión de insectos asociados a la descomposición de cuerpos en dos ecosistemas acuáticos en la Amazonia Andina. Metodología: Tres cadáveres fueron ubicados en una quebrada y tres en un lago, los muestreos se realizaron cada cinco horas desde el momento de la muerte hasta su esqueletización. Resultados: La descomposición total duró 14 días en la quebrada y 16 en el lago. Seis estadios de descomposición fueron observados: fresco sumergido; flotación temprana; flotación tardía; deterioro de la flotación; restos flotantes y restos sumergidos. En total se colectaron 1172 individuos en la quebrada y 1585 en el lago. Los gremios tróficos identificados fueron: trituradores; colectores; depredadores; necrófagos; sarcosaprófagos e incidentales. En la quebrada, los trituradores tricópteros fueron los más representativos en la etapa fresco sumergido. En el lago los trituradores, moscas de mayo y los depredadores hemípteros y odonatos fueron los más representativos en la etapa de flotación temprana. Los dípteros necrófagos y sarcosaprófagos fueron los más representativos en las etapas donde el cuerpo estuvo expuesto a la superficie en ambos ecosistemas. Conclusiones: El ISMP para ambos cadáveres fue similar en los dos hábitats, cada uno evidenció seis etapas de descomposición, lo que confirma que la descomposición en los ecosistemas acuáticos es diferente a la observada para los entornos terrestres.

Abstract Objectives: Analyze the succession of insects associated with the decomposition of bodies in two aquatic ecosystems in the Andean Amazon. Scope: Provide information on the insects associated with bodies that decompose within aquatic habitats and the duration times of each decomposition phase as a tool for the determination of the post-mortem submerged interval (PMSI). Methodology: Three carcasses were placed in a stream and three in a lake, the sampling carried out every five hours, from the moment of death to its skeletonization. Main results: Total decomposition lasted for 14 days in the stream and 16 days in the lake. Six decomposition stages were observed: submerged fresh, early floating, floating decay, bloated deterioration, floating remains and sunken remains. A total of 1172 organisms were collected on the carcasses in the stream and 1585 in the lake. Trophic relationships identified were: Shredders, collectors, predators, necrophages, sarcosaprophages and incidentals. In the stream, the caddisfly shredders were the most representative organisms in the submerged fresh stage. In the lake, the mayfly shredders, and hemipterous and odonate predators, were the most representative organisms in the early floating stage. The necrophagous and sarcosaprophagous diptera were the most representative organisms in the stages exposed to the surface in both ecosystems. Conclusions: The PMSI for both carcasses was similar in the two habitats, each showing six stages of decomposition, confirming that decomposition in lotic and lentic ecosystems is different from that observed for terrestrial environments.

RESUMO A entomologia forense é uma ferramenta frequentemente utilizada para estimar o intervalo de tempo entre a morte e a descoberta do corpo. Na área rural do município de Florencia, Caquetá, foi monitorada a sucessão de artrópodes associados à decomposição cadavérica, utilizando como modelo de estudo os cadáveres de três porcos (Sus scrofa). A amostragem de insetos e coleta de dados ambientais foram realizadas a cada cinco horas. A duração total do processo desde a morte até a esqueletização foi de 545 horas (22,7 dias). Foram coletados 30833 espécimes de insetos, distribuídos em nove ordens, 46 famílias, 95 gêneros e 106 espécies. Diptera foi o grupo mais representativo, com 23215 indivíduos (75,3%), seguido de Coleoptera, com 3711 indivíduos (12%) e Hymenoptera, com 3154 indivíduos (10,2%). Os estágios imaturos de Cochliomyia macellaria, Chrysomya albiceps, Hemilucilia semidiaphana e Ophyra aenescens foram as principais espécies envolvidas no consumo de tecidos e na aceleração do processo de decomposição. Devido à presença de formigas Cheliomyrmex sp., Camponotus sp. e Dinoponera sp. e de besouros Hister sp., Acylophorus sp. e Philonthus spp., não foi possível obter massas de ovos de Diptera suficientes para a criação de espécies colonizadoras. Nosso estudo fornece subsídios para determinar o intervalo post-mortem em investigações policiais e promover a entomologia forense em uma área rural da Amazônia Andina na Colômbia.

ABSTRACT Forensic entomology is a frequently used tool to estimate the time interval between death and the discovery of the corpse. Succession of arthropods associated with cadaveric decomposition was monitored in a rural area of the Municipality of Florencia, Department of Caquetá, Colombia. Three pigs (Sus scrofa) were used as study models. Insect sampling, and monitoring of carcasses and environmental conditions were carried out every five hours. The total time from death to skeletonization was of 545 hours (22.7 days). A total of 30833 insect individuals were collected. Specimens were distributed in nine orders, 46 families, 95 genera and 106 species. Diptera was the most abundant, with 23215 individuals (75.3%), followed by Coleoptera, with 3711 individuals (12%), and Hymenoptera, with 3154 individuals (10.2%). Immature stages of Cochliomyia macellaria, Chrysomya albiceps, Hemilucilia semidiaphana and Ophyra aenescens were the main species involved in tissue consumption and acceleration of the decomposition process. Due to the presence of ants Cheliomyrmex sp., Camponotus sp. and Dinoponera sp., and coleopterans Hister sp., Acylophorus sp. and Philonthus spp., it was not possible to obtain sufficient Diptera egg masses for rearing the colonizing species. These results can be used as a standard to determine the postmortem interval in criminal investigations in the rural area of the Andean Amazon, Caquetá, Colombia.

La entomología forense es una disciplina que utiliza insectos para obtener información útil en la determinación del intervalo postmortem (IPM). Las moscas de la familia Calliphoridae son muy utilizadas en entomología forense, sin embargo, su identificación a nivel de especie puede dificultarse cuando el individuo se encuentra incompleto o en estadio inmaduro. En el presente trabajo, se evaluó el potencial de la región ITS2 del genoma nuclear para la identificación de especies de Calliphoridae en Colombia utilizando tres aproximaciones: comparando distancias genéticas utilizando la metodología de códigos de barra, haciendo una reconstrucción filogenética, y con enzimas de restricción (PCR-RFLPs). Se secuenciaron un total de 520 pb en 44 individuos pertenecientes a 16 especies. Se calcularon los valores de distancia intraespecífica e interespecíficas utilizando el modelo K2P. Los valores de distancia intraespecífica oscilaron entre 0 y 0,252 %, mientras que las distancias interespecíficas fluctuaron entre 3,6 y 18,9 %, evidenciándose que esta técnica puede ser utilizada como código de barras genético en la identificación de especies de la familia Calliphoridae. Tanto en los análisis de Neighbour-Joining como en los análisis bayesianos el 90 % de los géneros presentan una monofilia sustentada en probabilidad posterior de 0,89 a 1. En todos los casos la especie Blepharicnema splendens agrupa con el género Lucilia. Con base en las secuencias obtenidas se utilizó la aplicación NEBCutter para identificar cuatro enzimas de restricción las cuales se probaron en el laboratorio y se comprobó su utilidad para la identificación rápida de especies de Calliphoridae en Colombia.

Forensic entomology is a discipline that uses insects to obtain useful information for the determination of the postmortem interval (PMI). Flies of the family Calliphoridae are extensively used for this purpose, however, the identification of these flies can be difficult when the individual is not an adult or when it is incomplete. In the present work, we tested the utility of the ITS2 region of the nuclear genome for the identification of Calliphoridae species in Colombia using three approaches: comparing genetic distances using the barcoding methodology, with a phylogenetic reconstruction, and with PCR-RFLPs. We sequenced 520 bp in 44 individuals belonging to 16 species of califorids. Intraspecific and interspecific distance values were calculated using the K2P model. The intraspecific distance values ranged between 0 and 0.252 %, while the interspecific distance values ranged between 3.6 and 18.9 %, indicating that this gene can be used as a genetic barcode for the identification of species of the Calliphoridae family. Both the Neighbour-Joining and Bayesian analyses recovered 90 % of the genera as monophyletic, with pp values between 0.89 and 1. Blepharicnema splendens was always recovered within the Lucilia genera. Based on the obtained sequences we used the NEBCutter application to identify four restriction enzymes that cut in a differential way and generated useful patterns for the identification of the species. The enzymes were successfully tested and confirmed the utility of this technique as a fast way to identify species of Calliphoridae in Colombia.