Characterising four Sarconesiopsis magellanica (Diptera: Calliphoridae) larval fat body-derived antimicrobial peptides

BACKGROUND The inappropriate use of antibiotics has led to the accelerated growth of resistance to antibiotics. The search for new therapeutic strategies (i.e., antimicrobial peptides-AMPs) has thus become a pressing need. OBJECTIVE Characterising and evaluating Sarconesiopsis magellanica larval fat body-derived AMPs. METHODS Fat body extracts were analysed by reversed-phase high-performance liquid chromatography (RP-HPLC); mass spectrometry was used for characterising the primary structure of the AMPs so found. ProtParam (Expasy) was used for analysing the AMPs' physico-chemical properties. Synthetic AMPs' antibacterial activity was evaluated. FINDINGS Four new AMPs were obtained and called sarconesin III, IV, V and VI. Sarconesin III had an α-helix structure and sarconesins IV, V and VI had linear formations. Oligomer prediction highlighted peptide-peptide interactions, suggesting that sarconesins III, V and VI could form self-aggregations when in contact with the microbial membrane. AMPs synthesised from their native molecules' sequences had potent activity against Gram-positive bacteria and, to a lesser extent, against Gram-negative and drug-resistant bacteria. Sarconesin VI was the most efficient AMP. None of the four synthetic AMPs had a cytotoxic effect. MAIN CONCLUSIONS S. magellanica larval fat body-derived antimicrobial peptides are an important source of AMPs and could be used in different antimicrobial therapies and overcoming bacterial resistance.

Species Identification of Sarcosaprophagous Flies Based on Vein Digital Image Analysis / 法医学杂志

Objective To identify species of common sarcosaprophagous flies based on digital image analysis of veins, in order to provide new idea for fast and accurate species identification of sarcosaprophagous flies in forensic entomology. Methods Random trapping of 226 male and female sarcosaprophagous flies that comprised of 7 common species, including Sarcophaga peregrina, Parasarcophaga ruficornis, Sarcophaga dux, Seniorwhitea reciproca, Bercaea cruentata, Aldrichina grahami, and Synthesiomysia nudiseta with carrion in the field was conducted. The 17 landmarks on the right wing of each fly were digitally processed and the images were analyzed. The effects of allometry were evaluated using a permutation test. Wing shape variations among 7 sarcosaprophagous fly species and female species was analyzed using canonical variate analysis （CVA）. Additionally, cross-validation test was used to evaluate the reliability of classification. Results Among 7 sarcosaprophagous fly species and female species, the effect of allometry had statistical significance （P<0.05）. The CVA results showed that among 7 sarcosaprophagous fly species and female species, differences in the wing shape were significant, and the first two canonical variates accounted for 82.9% and 84.1% of the total variation of vein shape. Vein digital image analysis can be used to separate the 7 common sarcosaprophagous flies, with an overall species identification accuracy of 81.2%-100.0%, and with a species identification accuracy of 75.0%-100.0% to distinguish the female flies of the 7 sarcosaprophagous flies species. Conclusion Vein digital image analysis is a relatively convenient and reliable method for identification of insect species, which can be used for species identification of common sarcosaprophagous flies.

Desenvolvimento pós-embrionário de Chrysomya putoria (Calliphoridae) em carne equina e sua importância para a entomologia e veterinária forenses / [Post embryonic development of Chrysomya putoria (Calliphoridae) using horse meat and its importance for forensic entomology and veterinary]

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.(AU)

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.(AU)

New records of Muscidae (Diptera) in Campo Grande, MS, Brazil / Novos registros de dípteros muscóides em Campo Grande, MS, Brasil

Synanthropic flies outstanding beside other flies due their relative abundance close to domestic animals and human population, to which they are able to cause myiasis or transmit pathogenic agents. As they're necrophagous they act as corpse decomposers and are useful in the forensic entomology in the post mortem interval determination. This study aimed to know flies diversity and abundance in Campo Grande, State of Mato Grosso do Sul. Captures were made weekly, utilizing three traps baited with decaying fish meat, from June of 2008 to May of 2009 in a remainder ciliary forest of the Embrapa's Cattle Beef Experimental Farm. The dipterans families that were considered and respectively number of collected specimens were: Calliphoridae (105,334); Muscidae (27,999); Sarcophagidae (21,083); Fanniidae (17,759) and Mesembrinellidae (305), totalizing 172,480 dipterous. To the local known species some Muscidae were increased as follows: Neomuscina atincticosta, Pseudoptilolepis elbida, Polietina orbitalis, Polietina flavithorax, Scutellomusca scutellaris, Graphomya analis and Morellia couriae.(AU)

As moscas sinantrópicas se destacam pelo fato de serem relativamente abundantes junto à população humana e animais domésticos, podendo causar a estes miíases ou transmitir agentes patogênicos. Por serem necrófagas, atuam na decomposição de cadáveres e são úteis para a entomologia forense como indicadores na determinação do intervalo post mortem - IPM. Este trabalho visou conhecer a diversidade e a abundância das espécies em Campo Grande, Estado de Mato Grosso do Sul. As capturas foram realizadas semanalmente, com três armadilhas, utilizando isca de peixe deteriorado, durante o período de junho de 2008 a maio de 2009 em mata ciliar remanescente na fazenda experimental da Embrapa Gado de Corte. Foram consideradas as seguintes famílias de dípteros, seguidas pelo número de exemplares obtidos: Calliphoridae (105.334); Muscidae (27.999); Sarcophagidae (21.083); Fanniidae (17.759) e Mesembrinellidae (305), totalizando 172.480 dípteros. Foram acrescentadas às espécies de Muscidae já notificadas para o local deste estudo: Neomuscina atincticosta, Pseudoptilolepis elbida, Polietina orbitalis, Polietina flavithorax, Scutellomusca scutellaris, Graphomya analis e Morellia couriae.(AU)

Caracterización citogenética de lucilia sericata (Meigen, 1826) (Diptera: calliphoridae), Cepa Bogotá, Colombia / Cytogenetic characterization of lucilia sericata (Meigen, 1826) (diptera: calliphoridae), Bogotá-Colombia strain / Caracterização citogenética de lucilia sericata (Meigen, 1826) (diptera: calliphoridae), cepa Bogotá, Colômbia

Introducción: Lucilia sericata es una especie de importancia médica y forense, utilizada en terapia larval para curar heridas crónicas y en estudios médico-legales empleada en la estimación del intervalo post mórtem y el traslado de cadáveres. No existen registros de las características citogenéticas de esta mosca en el neotrópico. El objetivo principal de este trabajo fue identificar las características morfométricas cromosómicas y las estructuras primarias del cariotipo, a partir de especímenes de L. sericata de la cepa Bogotá, Colombia. Materiales y métodos: Se tomaron huevos embrionados, que fueron previamente esterilizados en su superficie, se maceraron y luego fueron sembrados en el medio de cultivo L-15, suplementado con 20% de SFB, e incubados a una temperatura de 28 °C, sin atmosfera de C0(2). La preparación de los cromosomas se obtuvo de monocapas celulares semiconfluentes, empleando diversas soluciones: antimitótica (Colchicina), hipotónica (KCl 0,075 M) y fijadora (Carnoy: metanol y ácido acético; 3:1). Se llevó a cabo la técnica de bandeo C para la identificación de regiones cromosómicas de heterocromatina constitutiva. Resultados: Se obtuvieron parámetros morfométricos de cada par cromosómico. El número diploide del cariotipo obtenido de los cultivos celulares fue 2n = 12; éstos se clasificaron morfológicamente, de acuerdo con patrones previamente establecidos, así: los pares I, II, IV y V fueron metacéntricos, y el par III fue submetacéntrico. A su vez, el par sexual fue heteromórfico, siendo el cromosoma X metacéntrico y el cromosoma Y submetacéntrico. El bandeo C fue positivo para todos los pares cromosómicos. Conclusiones: Se establecieron las características citogenéticas de L. sericata, cepa Bogotá, Colombia, relacionadas con número, forma, tamaño, posición del centrómero y regiones heterocromáticas de los cromosomas.

Objective: Lucilia sericata is an important species for medical and forensic purposes, it is used in maggot therapy in the treatment of chronic wounds and in medical-legal studies for establishing the post-mortem interval and the transfer of corpses. Currently there are no records of the cytogenetic characteristics of this fly in Neotropical region. The main objective of this study was to identify morphometric characteristics and primary structures from karyotype of L. sericata strain Bogota, Colombia. Methods and materials: Embryonated eggs were taken, which were previously surface sterilized, macerated and then seeded in L-15 medium culture, supplemented with 20% FBS and incubated at 28 °C, without C0(2) atmosphere. The preparation of chromosomes was obtained from semiconfluent monolayers, pretreated with various solutions: antimitotic (Colchicine), hypotonic (KCl 0.075 M) and fixative (Carnoy, methanol and acetic acid, 3:1). C-banding technique was carried out to identify chromosomal regions of constitutive heterochromatin. Results: Morphometric parameters were obtained from each pair of chromosomes. The diploid karyotype number obtained from cell cultures was 2n = 12; they were classified morphologically, according to patterns established previously, as follows: pairs I, II, IV and V were metacentric and pair III was submetacentric. On the other hand, the sexual pair was heteromorphic, being X chromosome metacentric and Y chromosome submetacentric. C banding was positive for all chromosome pairs. Conclusions: The cytogenetic characteristics of L. sericata, strain Bogotá, were established according to number, shape, centromer position and heterochromatic regions.

Introdução: Lucilia sericata é uma espécie de importência médica e forense, utilizada em terapia larval para curar feridas crônicas e em estudos médico-legais empregada na estimação do intervalo post morteme o traslado de cadáveres. Não existem registros das características citogéneticas desta mosca no neotrópico. O objetivo principal deste trabalho foi identificar as características morfométricas cromossômicas e as estruturas primárias do cariótipo, a partir de especímenes de L. sericata da cepa Bogotá, Colômbia. Materiais e métodos: Tomaram-se ovos embrionados, que foram previamente esterilizados em sua superfície, se maceraram e depois foram semeados no médio de cultivo L-15, suplementado com 20% de SFB, e incubados a uma temperatura de 28 °C, sem atmosfera de C02. A preparação dos cromossomas obteve-se de monocamadas celulares semiconfluentes, utilizando diversas soluções: antimitótica (Colchicina), hipotônica (KCl 0,075 M) e fixadora (Carnoy: metanol y ácido acético; 3:1). Levou-se a cabo a técnica de bandas C para a identificação de regiões cromossômicas de heterocromatina constitutiva. Resultados: Se obtiveram parâmetros morfométricos de cada par cromossômico. O número diplóide do cariótipo obtido dos cultivos celulares foi 2n = 12; estes se classificaram morfologicamente, de acordo com patrões previamente estabelecidos, assim: os pares I, II, IV e V foram metacêntricos, e o par III foi submetacéntrico. Por sua vez, o par sexual foi heteromórfico, sendo o cromossoma X me-tacêntrico e o cromossoma Y submetacêntrico. As bandas C foram positivas para todos os pares cromossômicos. Conclusões: Se estabeleceram as características citogenéticas de L. sericata, cepa Bogotá, Colômbia, relacionadas com número, forma, tamanho, posição do centrômero e regiões heterocromáticas dos cromossomas.

Diversidade de Calliphoridae (Insecta: Diptera) na base de extração petrolífera da Bacia do Rio Urucu, na Amazônia brasileira / Diversity of Calliphoridae flies (Insecta: Diptera) in the oil extraction field of Rio Urucu basin, in Brazilian Amazonian

A diversidade e abundância de dípteros califorídeos de três ambientes (clareira artificial, clareira natural e mata) de Porto Urucu/AM foram avaliadas em coletas anuais realizadas em 2004, 2005 e 2006. Ao longo destes três anos foram coletados 2.121 exemplares pertencentes a 14 espécies. As espécies mais abundantes foram Chloroprocta idioidea (Robineau- Desvoidy), Eumesembrinella randa (Walker) e Hemilucilia semidiaphana (Rondani). Os habitats de matas e clareiras naturais apresentaram maior abundância de califorídeos quando comparados às clareiras artificiais, com índices de diversidade e equitabilidade também maiores do que em clareiras artificiais, onde a dominância foi mais elevada.

The diversity and abundance of blow flies in three environments (anthropic gaps, tree-fall gaps, and primary "terra firme" forest) of Porto Urucu/AM were evaluated in annual collects realized in 2004, 2005, and 2006. During these three years were collected 2,121 specimens belonging to 14 species. The most abundant species were Chloroprocta idioidea (Robineau- Desvoidy), Eumesembrinella randa (Walker), and Hemilucilia semidiaphana (Rondani). Primary forests and tree-fall gaps enviroments, showed higher blow flies abundance than anthropic gaps, with diversity and equitability index higher than anthropic gaps, when the dominance value was high.

Notes on necrophagous flies (Diptera: Calyptratae) associated to fish carrion in Colombian Amazon / Notas sobre moscas necrófagas (Diptera: Caliptratae) associadas a carcaças de peixe na Amazônia Colombiana

In recent years, there has been an increasing number of studies on carrion fly communities due to their medical importance and as a consequence of the large number of studies on forensic entomology. Surprisingly few studies have adressed with the asynantropic flies of the Amazon, and none were done in Colombia. A faunistic study of asynantropic flies of the families Calliphoridae, Sarcophagidae, Muscidae and Fannidae in three different landscapes of the Colombian Amazon is presented, trapping effectiveness is assessed, and the first record of Mesembrinella batesi (Aldrich, 1922) and Fannia femoralis (Stein, 1897) from Colombia is reported.

Apesar de existir uma quantidade considerável de estudos sobre dípteros decompositores devido a sua importância medica e ao avanço da entomologia forense, poucos dizem respeito as moscas asinantrópicas na Amazônia e nada foi feito na Colômbia. No presente trabalho é feito um estudo faunístico sobre moscas, principalmente das famílias Calliphoridae, Sarcophagidae, Muscidae e Fanniidae em três diferentes paisagens da Amazônia Colombiana, além do primeiro registro das espécies Mesembrinella batesi (Aldrich, 1922) e Fannia femoralis (Stein, 1897) para Colômbia e avaliação da amostragem utilizada.

Estudio de cultivos celulares primarios derivados de Lucilia sericata (Diptera: Calliphoridae) / Study of primary cell cultures derived from Lucilia sericata (Diptera: Calliphoridae)

El propósito principal de la investigación aquí presentada fue obtener cultivos celulares primarios derivados de Lucilia sericata (Diptera: Calliphoridae). Esta mosca necrófaga es utilizada para determinar el intervalo post-mortem y en terapia larval. A partir de huevos embrionados, se realizaron explantes en diversos medios de cultivo (Grace, Schneider, MM/VP12, DMEM, Grace/L-15 y L-15), suplementados con 20% de suero fetal bovino. La esterilización del material biológico se efectuó mediante la aplicación de soluciones de formaldehido e hipoclorito de sodio. El crecimiento celular se inicio en los medios L-15, MM/VP12, Grace/L-15 y Schneider, en un tiempo promedio de 10 días después de efectuadas las siembras de tejidos embrionarios, mediante la proliferación de grupos de colonias dispersas en la superficies de los frascos de cultivo y a partir de las terminaciones de los fragmentos larvales. La evolución del crecimiento celular hasta la formación de la monocapa semiconfluente fue relativamente rápida, se alcanzo a las tres semanas post-explantes. La morfología de las células en los cultivos fue heterogénea, se destacaron formas epitelioides, similares a nerviosas, gigantes e irregulares. La comparación de las características de crecimiento de los cultivos celulares de L. sericata con los obtenidos de otras especies de dípteros mostro mayor favorabilidad en la evolución, en razón a que las células se adaptaron mejor a las condiciones fisico-quimicas de varios medios de cultivo. Este es el primer informe de cultivos celulares de una mosca de la familia Calliphoridae.

The main purpose of this study was to obtain primary cell cultures derived from Lucilia sericata (Diptera: Calliphoridae). Necrophagous this fly is used for determination of post-mortem interval and larval therapy. Since explants embryonated eggs were performed in various culture media (Grace Schneider, MM/VP12, DMEM, Grace/L-15 and L-15), supplemented with 20% fetal serum. Sterilization of the biological material was carried out by immersing it in formaldehyde and sodium hypochlorite solutions. The cell growth was initiated in the L-15, MM/VP12, and Schneider Grace/L-15 in an average time of 10 days after completion of planting by the proliferation of groups of colonies scattered on the surface of the boxes crops and also from the endings of larval fragments. The evolution of cell growth to the formation of monolayer semi-confluent was relatively fast, reaching at 3 weeks post-explant. Cellular morphology in cultured cells was heterogeneous, especially epithelioid forms, similar to nerve, giant and irregular. Comparison of the growth characteristics of these cell cultures with those obtained from other species of flies was more favorable in the evolution of those obtained from L. sericata, on the grounds that the cells are better adapted to the physical-chemical conditions of several culture media. This is the first report of a cell culture-fly family Calliphoridae.

A new record of Bengalia emarginata Malloch, 1927 (Diptera: Calliphoridae) from Malaysia

During a forensic entomological study conducted at an oil palm plantation in Tanjung Sepat, Kuala Langat, Selangor, a Bengalia emarginata Malloch, 1927 (Diptera: Calliphoridae: Calliphorinae: Bengalini) was collected for the first time. Two adults were collected nearby the pig carcass by the first author and identified by the second. Prior to this finding, nine species of Bengalia were recorded from peninsular Malaysia or Borneo. Male of B. emarginata are different from Bengalia varicolor Fabricious by the following characters: Sternite 5 projection rounded with small identation and mid tibia double-fringed in ventral surface.