Primeros registros de cuatro especies de Calliphoridae (Diptera: Oestroidea) de Yucatán, México

La familia Calliphoridae es un componente integral de la estimación del intervalo post mortem (IPM) debido a su desarrollada capacidad para detectar la descomposición de cadáveres a grandes distancias y la habilidad para colonizarlos a diferentes temperaturas en diversas áreas geográficas. Se reportan cuatro primeros registros de la familia Calliphoridae para la entomofauna de Yucatán, México: Chrysomya putoria Wiedemann, Chrysomya rufifacies Macquart, Lucilia cuprina Wiedemanny y L. sericata Meigen. Se incluye una clave de identificación e ilustraciones para las nueve especies presentes en el estado y comentarios sobre su distribución.

Maggot debridement therapy to treat hard-to-heal diabetic foot ulcers: a single-centre study

Objective: Maggot debridement therapy (MDT) has seen a resurgence in recent years in the treatment of hard-to-heal wounds, as a result of rising antibiotic resistance. The sterilised larvae of Lucilia cuprina have been used in MDT in Malaysia since 2003, with encouraging results for the treatment of hard-to-heal diabetic wounds. We report a case series of 30 patients selected from our clinic by convenient sampling with diabetic lower limb ulcers treated with MDT. The average age of patients receiving MDT was >50 years. Of the 30 patients in the study, nine were female and 21 were male. All patients had underlying diabetes, two patients had leg ulcers and 28 patients had diabetic foot ulcers. Sterilised Lucilia cuprina larvae were applied via a standard method of 10 maggots per square centimetre and dressed with sterile gauze. The study endpoint was defined as ≤5% coverage with slough or necrotic tissue following three successive applications of MDT. In this study, maximum debridement of wounds was achieved in 96.6% (29 patients) of our patients, with ≤5% coverage with slough or necrotic tissue, in addition to a reduction in wound-related pain, as assessed by a visual analogue scale. No adverse events were reported. The findings of this study support the use of MDT as a safe, efficacious, and cost-effective method of managing diabetic wounds.

Proteomic Analysis of Glutathione Transferases from Lucilia Cuprina

<p>The glutathione transferases are a family of multifunctional enzymes involved in detoxification of xenobiotic and endogenous electrophilic compounds. Interest in insect GSTs has primarily focused on their role in insecticide resistance. The sheep blowfly, Lucilia cuprina is a major economic problem for the sheep meat and wool industries in Australasia and hence this thesis has attempted the study of the Lucilia cuprina GST family, using proteomics, with a view to eventually determining their role in insecticide resistance. Combinations of different affinity matrices (glutathione-Sepharose matrix (GSH) followed by dinitrophenyl-glutathione-Sepharose matrix (DNP-GSH)) and two-dimensional electrophoresis has successfully isolated members from major four insect GST classes: Sigma, Delta, Epsilon and Omega. Drosophila melanogaster has been used as a model insect throughout as a basis for comparison. To characterise Lucilia GSTs, the whole metazoan fragmentation database was used for sequence alignment with Lucilia peptides. This approach is broad and speculative but predicts a possible classification of the GSTs based on % similarity and % identity. This method of characterisation yielded match scores that provided a basis for classification, which must at present be regarded as tentative and in need of confirmation. In D. melanogaster and L. cuprina, GSH affinity-purified extracts showed the presence of only Sigma and Delta GSTs. In D. melanogaster, the DNP-GSH affinity-purified GSTs showed mostly the presence of Epsilon and Omega GSTs whereas in L. cuprina no Omega GSTs were detected. In both species, the migration pattern of Delta GST on 2D PAGE gel indicated possible post-translational modification. The results from analysis of LC-MS/MS data by the software PEAKS suggested deamidation at asparagine and glutamine residues in a limited number of the matched peptides of Delta GST. GST activity was present in all developmental stages of L. cuprina. The number of isoenzymes and their extent of expression vary as the insect develops. Delta GSTs were present in all developmental stages. The Sigma GST started expressing from the larval stage and was abundantly present in adult stage. The DNP-GSH affinity matrix purified GSTs which have been tentatively classified as Mu-like GSTs were present in egg, larvae and pupae but totally absent in adult stage. The GST families were characterised by proteomics in the main body sections of L. cuprina. Higher GST activity towards 1-chloro-2, 4-dinitrobenzene (CDNB) was found in the thorax (65.2 %) followed by the abdomen (19.6%) and the head (15.2%). The cytosolic GSTs of a resistant strain (PY81) of L. cuprina had significantly higher (2.26- and 2.6- fold) activity than the susceptible strains (NSW and CSIRO) towards CDNB and 2, 3-dichloro, 4-nitrobenzene (DCNB) respectively. The proteomic analysis of DNP-GSH purified extract from susceptible and resistant strains showed quantitatively higher expression of GSTs on 2D PAGE gel of the PY81 strain. The in vitro interaction of purified GSTs and model insecticides studied by high performance liquid chromatography revealed that Delta and DNP-GSH affinity-purified GSTs catalyse the conjugation of the insecticides to reduced glutathione but Sigma GST had almost no activity.</p>

Proteomic Analysis of Glutathione Transferases from Lucilia Cuprina

<p>The glutathione transferases are a family of multifunctional enzymes involved in detoxification of xenobiotic and endogenous electrophilic compounds. Interest in insect GSTs has primarily focused on their role in insecticide resistance. The sheep blowfly, Lucilia cuprina is a major economic problem for the sheep meat and wool industries in Australasia and hence this thesis has attempted the study of the Lucilia cuprina GST family, using proteomics, with a view to eventually determining their role in insecticide resistance. Combinations of different affinity matrices (glutathione-Sepharose matrix (GSH) followed by dinitrophenyl-glutathione-Sepharose matrix (DNP-GSH)) and two-dimensional electrophoresis has successfully isolated members from major four insect GST classes: Sigma, Delta, Epsilon and Omega. Drosophila melanogaster has been used as a model insect throughout as a basis for comparison. To characterise Lucilia GSTs, the whole metazoan fragmentation database was used for sequence alignment with Lucilia peptides. This approach is broad and speculative but predicts a possible classification of the GSTs based on % similarity and % identity. This method of characterisation yielded match scores that provided a basis for classification, which must at present be regarded as tentative and in need of confirmation. In D. melanogaster and L. cuprina, GSH affinity-purified extracts showed the presence of only Sigma and Delta GSTs. In D. melanogaster, the DNP-GSH affinity-purified GSTs showed mostly the presence of Epsilon and Omega GSTs whereas in L. cuprina no Omega GSTs were detected. In both species, the migration pattern of Delta GST on 2D PAGE gel indicated possible post-translational modification. The results from analysis of LC-MS/MS data by the software PEAKS suggested deamidation at asparagine and glutamine residues in a limited number of the matched peptides of Delta GST. GST activity was present in all developmental stages of L. cuprina. The number of isoenzymes and their extent of expression vary as the insect develops. Delta GSTs were present in all developmental stages. The Sigma GST started expressing from the larval stage and was abundantly present in adult stage. The DNP-GSH affinity matrix purified GSTs which have been tentatively classified as Mu-like GSTs were present in egg, larvae and pupae but totally absent in adult stage. The GST families were characterised by proteomics in the main body sections of L. cuprina. Higher GST activity towards 1-chloro-2, 4-dinitrobenzene (CDNB) was found in the thorax (65.2 %) followed by the abdomen (19.6%) and the head (15.2%). The cytosolic GSTs of a resistant strain (PY81) of L. cuprina had significantly higher (2.26- and 2.6- fold) activity than the susceptible strains (NSW and CSIRO) towards CDNB and 2, 3-dichloro, 4-nitrobenzene (DCNB) respectively. The proteomic analysis of DNP-GSH purified extract from susceptible and resistant strains showed quantitatively higher expression of GSTs on 2D PAGE gel of the PY81 strain. The in vitro interaction of purified GSTs and model insecticides studied by high performance liquid chromatography revealed that Delta and DNP-GSH affinity-purified GSTs catalyse the conjugation of the insecticides to reduced glutathione but Sigma GST had almost no activity.</p>

Wound healing and antibacterial activities of water-soluble chitosan nanoparticles and excretion/secretion as a natural combination from medicinal maggots, Lucilia cuprina

Wounds management takes a high interest in the medical field and the addition of antimicrobial agents in an assortment of wound dressings leads to delay the wound healing. This study aimed at preparing natural combination between excretion/secretion (ES) and water-soluble chitosan nanoparticles (from Lucilia cuprina maggots) and investigating its antibacterial and wound healing activities. ES of maggots was collected, and the water-soluble chitosan nanoparticles (WSCNPs) were prepared and characterized. Antibacterial activities of combinations were evaluated against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Proteus vulgaris. ES-WSC-2 combination that contains 50% ES and 1% WSCNPs showed highest antibacterial activity against all tested bacteria compared to the other combinations. In vitro, the ES-WSC-2 combination was used to study the wound healing activity by scratch assay. The synergism between ES and WSCNPs (in ES-WSC-2 combination) accelerated the wound healing rate which suggests the use of this combination as an effective natural antibacterial and wound healing agent.

Muscidae e Calliphoridae (Diptera, Brachycera) associados à decomposição cadavérica no noroeste do Paraná

O presente trabalho teve como objetivo listar quais são os dípteros necrófagos na região noroeste do Paraná. As informações fazem parte de um projeto, aprovado no comitê de ética em pesquisa de animais, em desenvolvimento no IFPR Campus Umuarama. Durante o estudo, um cadáver de suíno com o peso aproximado de 10 kg foi utilizado como substrato, o qual foi colocado em uma gaiola com uma tela, para impedir a entrada de vertebrados necrófagos. A coleta dos insetos foi realizada por meio de uma armadilha modificada. A gaiola de metal que continha o substrato estava fixada no terreno do IFPR Campus Umuarama. Em relação aos insetos foram coletadas cinco espécies de Muscidae (Sarcopromusca pruna; Musca domestica; Hydrotaea aenescens; Sythesiomyia nudiseta e Stomoxys calcitrans) e seis espécies de Calliphoridae (Cochliomyia macellaria; Chrysomya albiceps; Chrysomya megacephala; Lucilia eximia; Lucilia cuprina e Hemilucilia segmentaria). Os espécimes de Stomoxys calcitrans podem ter sido coletados devido à presença de gado de corte próximo ao local. O processo de decomposição se completou em seis dias, o que pode ser devido aos altos índices de temperatura e baixos índices de umidade observados no local. Nesse período foram observados quatro estágios: fresco; enfisematoso; coliquativo e esqueletização. Estes dados são importantes para a construção de um banco de dados de espécies de importância forense.

A Field Evaluation of the LuciTrap and the Western Australian Trap with Three Different Baits Types for Monitoring Lucilia cuprina and Lucilia sericata in New Zealand

Flytraps can be used on farms to monitor the populations of primary strike flies (Lucilia cuprina and Lucilia sericata) and, hence, offer a view regarding the incidence of flystrike on sheep. This study aimed to contrast the specificity and effectiveness of the LuciTrap with its combination of three chemical lures (Lucilures) and the Western Australian Trap with three bait types (LuciLure, Sheep liver with 30% sodium sulphide and squid). A mean model and rate model were fitted to the data. The mean model showed no difference (p > 0.05) in the mean weekly catch for L. cuprina between the Western Australian Trap with LuciLures and the Western Australian Trap baited with sheep liver with 30% sodium sulphide (p < 0.05). Whereas, for L. sericata, no difference (p > 0.05) was found between the Western Australian Trap with LuciLures, the Western Australian Trap baited with sheep liver with 30% sodium sulphide and the LuciTrap. The rate model illustrated that the Western Australian Trap with sheep liver with 30% sodium sulphide and LuciTrap did not differ (p > 0.05) for L. cuprina and L. sericata. Combined, these results indicate that New Zealand farmers can use either the LuciTrap or the Western Australian Trap with sheep liver with 30% sodium sulphide to monitor these target species.