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Fournier's Gangrene is a fulminating necrotizing fasciitis of the perineum and genitalia. Standard treatment involves immediate excision of all necrotic tissue, aggressive antibiotic coverage, and supportive medical care. Still, the infection is commonly fatal or disfiguring. Wound treatment with disinfected blowfly larvae (maggot debridement therapy or MDT) has been shown to be highly effective, with multiple studies demonstrating effective debridement, disinfection, and promotion of granulation tissue. MDT also has been associated with preservation of viable tissue and minimised blood loss. This report describes a prospective clinical study of MDT for Fournier's gangrene aimed to test the hypothesis that early use of maggots could decrease the number of surgical treatments required to treat Fournier's gangrene. Subjects were provided with one initial surgical excision, followed by debridement using only medical grade Lucilia sericata larvae. Only two subjects were enrolled, both diabetic men. Intensive care and culture-directed antimicrobial coverage were administered as usual. Maggot debridement was associated with the disappearance of necrotic tissue, control of infection and granulation tissue growth. In both subjects, wounds healed without requiring further surgical resection or anatomical reconstruction. Maggot therapy decreased the number of surgical procedures that otherwise would have been necessary, and led to favourable outcomes.

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This study's main objective was to evaluate the action of larval therapy derived from Lucilia sericata and Sarconesiopsis magellanica (blowflies) regarding Leishmania panamensis using an in vivo model. Eighteen golden hamsters (Mesocricetus auratus) were used; they were divided into 6 groups. The first three groups consisted of 4 animals each; these, in turn, were internally distributed into subgroups consisting of 2 hamsters to be used separately in treatments derived from each blowfly species. Group 1 was used in treating leishmanial lesions with larval therapy (LT), whilst the other two groups were used for evaluating the used of larval excretions and secretions (ES) after the ulcers had formed (group 2) and before they appeared (group 3). The three remaining groups (4, 5 and 6), consisting of two animals, were used as controls in the experiments. Biopsies were taken for histopathological and molecular analysis before, during and after the treatments; biopsies and smears were taken for assessing parasite presence and bacterial co-infection. LT and larval ES proved effective in treating the ulcers caused by the parasite. There were no statistically significant differences between the blowfly species regarding the ulcer cicatrisation parameters. There were granulomas in samples taken from lesions at the end of the treatments. The antibacterial action of larval treatment regarding co-infection in lesions caused by the parasite was also verified. These results potentially validate effective LT treatment against cutaneous leishmaniasis aimed at using it with humans in the future.

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Larval therapy is used as alternative treatment for hard-to-heal chronic and infected wounds. Lucilia sericata is the most used blowfly species. However, it has been shown recently that Sarconesiopsis magellanica larval excretions and secretions have potent antibacterial activity; this blowfly belongs to the Calliphoridae family. The present work has dealt with evaluating larval therapy using S. magellanica on wounds induced in diabetic rabbits and its action was compared to the effect induced by L. sericata. Twelve New Zealand White rabbits (Oryctolagus cuniculus) were used; they were divided into 4 groups, the first two being treated with larval therapy derived from both aforementioned necrophagous blowflies, an antibiotic was used in the third and the fourth was used as control. All the animals were wounded on the back and infected with Pseudomonas aeruginosa and Staphylococcus aureus. Samples of the secretion from each animal's infected wound were taken and sown on blood agar. The colony forming units were then counted. The PUSH scale was used for the macroscopic evaluation of the wounds. Bacterial control was encountered 48 h post-treatment in the treatments involving larval therapy and to a lesser extent with the antibiotic. Likewise, wound debridement was quicker and more efficient with larval therapy compared to the antibiotic group; however, wound closing time was 23 days in all treatments. The group treated with S. magellanica larvae had relatively quicker evolution until the proliferation phase and the start of maturation, even though there were no significant differences between both blowfly species evaluated here regarding treatments by the end of the treatment period. The present study has validated the diabetic rabbit model for inducing chronic wounds regarding larval therapy and has likewise confirmed the effectiveness of S. magellanica-derived larval therapy as an alternative for curing and healing wounds.

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Objetivo: Evaluar, en condiciones in vitro , la actividad antibacterial de los extractos de cuerpos grasos y de la hemolinfa de larvas de tercer estadio de Sarconesiopsis magellanica , la cual se comparó con los efectos obtenidos de las mismas sustancias derivadas de Lucilia sericata . S. magellanica (Diptera: Calliphoridae) es una mosca de importancia principalmente forense, utilizada en la determinación del intervalo post mortem . Por sus hábitos necrófagos, es considerada un modelo potencialmente útil en terapia larval. Material y métodos: Se extrajeron los cuerpos grasos de las larvas mediante la técnica de disección corporal y la hemolinfa se obtuvo mediante decapitación y centrifugación de los especímenes larvales. Las bacterias evaluadas fueron Staphylococcus aureus y Pseudomonas aeruginosa . Los métodos utilizados para evaluar la actividad antibacterial fueron difusión en agar y unidades formadoras de colonias (UFC/ml). Resultados: Después de la correspondiente incubación, los resultados generales mostraron que la actividad antibacterial de la hemolinfa y de los cuerpos grasos, tanto de L. sericata como de S. magellanica , fueron efectivos contra S. aureus y P. aeruginosa sin diferencias significativas entre las especies de moscas, aunque con algunas diferencias entre las cepas bacterianas. Conclusiones: Los resultados obtenidos sugieren que estas sustancias podrían tener un efecto similar en el tratamiento de heridas infectadas contra los microorganismos evaluados.

Objective: This study aimed to evaluate the in vitro antibacterial activity of fat body and hemolymph extracts from Sarconesiopsis magellanica (Diptera: Calliphoridae) third-instar larvae, compared to the effect obtained using the same extracts but derived from Lucilia sericata . S. magellanica blowflies are considered important in forensic sciences due to their usefulness in determining the post mortem interval. This blowfly could be useful in larval therapy due to its necrophagous habits. Materials and methods: Fat body from larvae was removed by dissection, and hemolymph via decapitation and centrifugation of larval specimens. The antibacterial effect was tested against Staphylococcus aureus and Pseudomonas aeruginosa using two methods: agar diffusion and colony forming units (CFU/mL). Results: Hemolymph and fat body extracts derived from both L. sericata and S. magellanica were effective against S. aureus and P. aeruginosa , with no significant differences between blowfly species, although with some differences between the bacterial strains. Conclusions: The results obtained suggest that S. magellanica and L. sericata fat body and hemolymph extracts might have a similar antimicrobial activity against these microorganisms when used to treat infected wounds.

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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.

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The objective of this work was to establish, under experimental laboratory conditions, a colony of Lucilia sericata, Bogotá-Colombia strain, to build life tables and evaluate two artifcial diets. This blowfy is frequently used in postmortem interval studies and in injury treatment. The parental adult insects collected in Bogotá were maintained in cages at 22°C±1 average temperature, 60 percent±5 relative humidity and 12 h photoperiodicity. The blowfies were fed on two artifcial diets that were evaluated over seven continuous generations. Reproductive and population parameters were assessed. The life cycle of the species was expressed in the number of days of the different stages: egg = 0.8±0.1, larvae I = 1.1±0.02, larvae II = 1.94±0.16, larvae III = 3.5±0.54, pupae = 6.55±0.47, male adult = 28.7±0.83 and female adult = 33.5±1.0. Total survival from egg stage to adult stage was 91.2 percent for diet 1, while for diet 2 this parameter was 40.5 percent. The lifetime reproductive output was 184.51±11.2 eggs per female. The population parameters, as well as the reproductive output of the blowfies that were assessed, showed relatively high values, giving evidence of the continuous increase of the strain over the different generations and making possible its maintenance as a stable colony that has lasted for more than two years.

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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.

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Introducción. Durante las últimas dos décadas, la terapia larval ha resurgido como una alternativa confiable y segura para la cura de úlceras cutáneas que no responden a los tratamientos convencionales. Objetivo. Evaluar el uso de las larvas de Lucilia sericata en el tratamiento de heridas infectadas con Pseudomonas aeruginosa en un modelo animal. Materiales y métodos. Se tomaron 12 conejos, los cuales fueron divididos al azar en 3 grupos homogéneos: al primero se le aplicó terapia larval, el segundo se trató con terapia antibiótica y el tercero fue establecido como control. A cada uno de los animales se les realizó una herida, luego se inoculó en ésta una suspensión de P. aeruginosa y, finalmente, al registrarse el desarrollo de la infección, se procedió, en los dos primeros grupos, a los tratamientos correspondientes. Para la evaluación macroscópica de las heridas, se tuvo en cuenta la presencia de edema y exudado, mal olor, inflamación alrededor de la herida y apariencia del tejido de granulación. Al proceso de cicatrización se le hizo seguimiento a través de una técnica dermohistológica. Resultados. Se registraron claras diferencias entre el grupo de animales tratados con terapia larval vs. el grupo tratado con terapia convencional de antibióticos, estableciéndose un periodo de 10 días para alcanzar la cicatrización en el grupo de terapia larval mientras que en el segundo grupo el proceso se cumplió en 20 días. Conclusiones. S e demostró la eficacia de las larvas de L. sericata en el tratamiento de heridas infectadas con P. aeruginosa.

Introduction. During the last two decades the larval therapy has reemerged as a safe and reliable alternative for the healing of cutaneous ulcers that do not respond to the conventional treatments. Objective. To evaluate the use of the larvae of Lucilia sericata as a treatment for infected wounds with Pseudomonas aeruginosa in an animal model. Materials and methods. Twelve rabbits were randomly distributed in 3 groups: the first group was treated with larval therapy; the second was treated with antibiotics therapy and to the third no treatment was applied, therefore was established as a control group. To each animal a wound was artificially induced, and then a suspension of P. aeruginosa was inoculated into the lesion. Finally, every rabbit was evaluated until the infection development was recognized and treatment was set up for the first two groups according with the protocols mentioned above. Macroscopic evaluation of the wounds was based on the presence of edema, exudates, bad odor, inflammation around the wound and the presence of granulation tissue. The healing process was evaluated by monitoring histological changes in the dermal tissue. Results. Differences in the time required for wound healing were observed between the first group treated with larval therapy (10 days) and the second group treated with conventional antibiotics therapy (20 days). Conclusion. The L. sericata larva is and efficient tool as a therapy for infected wounds with P. aeruginosa.

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The objective of this study was to determine synathropic index of adult Calliphoridae (Diptera) collected in the city of Valdívia, Chile (39°48 S; 73°15 ' W). For this, samples were taken monthly between September 1996 and August 1997 in three different localities representing urban, rural and natural environments. Rat carcass, fish , and chicken viscera were used as baits. Four blowfly species were collected: Calliphora vicina Robineau-Desvoidy, the dominant species in all three baits and sites of capture, followed by Lucilia sericata (Meigen), both species being eusynanthropic. The other two species, Compsomyiops fulvicrura (Robineau-Desvoidy) and Sarconesia magellanica (Le Guillou), were collected in smaller numbers and were characterized respectively as hemysinanthropic and asynanthropic. In addition to the synanthropic index, the monthly abundance and the efficiency of the baits as attractans to the flies as well as their medical and sanitary importance were also determined and discussed.

O objetivo do presente trabalho foi determinar o índice de sinantropia de adultos de Calliphoridae (Diptera) coletados na cidade de Valdívia, Chile (39°48' S; 73°15' W). As moscas foram coletadas mensalmente entre setembro de 1996 e agosto de 1997 em três localidades diferentes, representando os ambientes urbano, rural e natural. Carcaças de ratos albinos, vísceras de peixe e de galinha foram usadas como iscas. Quatro espécies de Calliphoridae foram capturadas: Calliphora vicina Robineau-Desvoidy a espécie dominante nas três iscas e nos três locais de coleta, seguida por Lucilia sericata (Meigen). Ambas as espécies foram consideradas como sinantrópicas. As outras duas espécies, Compsomyiops fulvicrura (Robineau-Desvoidy) e Sarconesia magellanica (Le Guillou), forams coletadas em menor número tendo sido consideradas respectivamente como hemissinanstrópica e assinantrópica. Além do índice de sinantropia, foram também determinadas a abundância mensal e a eficiência das iscas na atração das moscas, assim como foi avaliada a importância médico-sanitária das moscas.