Curva de sobrevivência e estimativa de entropia em Lucilia cuprina (Diptera, Calliphoridae)

Lucilia cuprina (Wiedemann, 1830) is a cosmopolite blowfly species of medical and veterinary importance because it produces myiasis, mainly in ovine. In order to evaluate the demographic characteristics of this species, survivorship curves for 327 adult males and 323 adult females, from generation F1 maintained under experimental conditions, were obtained. Entropy was utilized as the estimator of the survival pattern to quantify the mortality distribution of individuals as a function of age. The entropy values 0.216 (males) and 0.303 (females) were obtained. These results denote that, considering the survivorship interval until the death of the last individual for each sex, the males present a tendency of mortality in more advanced age intervals, in comparison with the females.

Dispersão larval radial pós-alimentar em Lucilia cuprina (Diptera, Calliphoridae): profundidade, peso e distância de enterramento para pupação

Blowflies utilize discrete and ephemeral sites for breeding and larval nutrition. After the exhaustion of food, the larvae begin dispersing to search for sites to pupate or for additional food source, process referred as postfeeding larval dispersal. Some aspects of this process were investigated in Lucilia cuprina (Wiedemann, 1830), utilizing a circular arena to permit the radial dispersion of larvae from the food source in the center. To determine the localization of each pupa, the arena was split into 72 equal sectors from the center. For each pupa, distance from the center of arena, weight and depth were determined. Statistical tests were performed to verify the relation among weight, depth and distance of burying for pupation. It was verified that the larvae that disperse farthest are those with lowest weights. The majority of individuals reached the depth of burying for pupation between 7 and 18 cm. The study of this process of dispersion can be utilized in the estimation of postmortem interval (PMI) for human corpses in medico-criminal investigations.

Dispersão larval radial pós-alimentar em Lucilia cuprina (Diptera, Calliphoridae): profundidade, peso e distância de enterramento para pupação

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Curva de sobrevivência e estimativa de entropia em Lucilia cuprina (Diptera, Calliphoridae)

Lucilia cuprina (Wiedemann, 1830) is a cosmopolite blowfly species of medical and veterinary importance because it produces myiasis, mainly in ovine. In order to evaluate the demographic characteristics of this species, survivorship curves for 327 adult males and 323 adult females, from generation F1 maintained under experimental conditions, were obtained. Entropy was utilized as the estimator of the survival pattern to quantify the mortality distribution of individuals as a function of age. The entropy values 0.216 (males) and 0.303 (females) were obtained. These results denote that, considering the survivorship interval until the death of the last individual for each sex, the males present a tendency of mortality in more advanced age intervals, in comparison with the females.

Baseline laboratory bioassay data for spinosad against populations of Australian sheep blowfly, Lucilia cuprina (Wiedemann) (Diptera : Calliphoridae)

Spinosad was an effective larvicide against the Australian sheep blowfly, Lucilia cuprina. A survey of 41 field populations indicated no cross-resistance to spinosad from existing organophosphate resistance. The data presented serve as baseline data for future resistance surveys.

Ecologia populacional de Lucilia cuprina (Wiedemann, 1830) (Diptera, Calliphoridae): aspecto do desenvolvimento em diferentes densidades larvais e seus efeitos sobre os adultos resultantes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The influence of temperature on the behavior of burrowing in larvae of the blowflies, Chrysomya albiceps and Lucilia cuprina, under controlled conditions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The gene transformer-2 of Anastrepha fruit flies (Diptera, Tephritidae) and its evolution in insects

Background: In the tephritids Ceratitis, Bactrocera and Anastrepha, the gene transformer provides the memory device for sex determination via its auto-regulation; only in females is functional Tra protein produced. To date, the isolation and characterisation of the gene transformer-2 in the tephritids has only been undertaken in Ceratitis, and it has been shown that its function is required for the female-specific splicing of doublesex and transformer pre-mRNA. It therefore participates in transformer auto-regulatory function. In this work, the characterisation of this gene in eleven tephritid species belonging to the less extensively analysed genus Anastrepha was undertaken in order to throw light on the evolution of transformer-2. Results: The gene transformer-2 produces a protein of 249 amino acids in both sexes, which shows the features of the SR protein family. No significant partially spliced mRNA isoform specific to the male germ line was detected, unlike in Drosophila. It is transcribed in both sexes during development and in adult life, in both the soma and germ line. The injection of Anastrepha transformer-2 dsRNA into Anastrepha embryos caused a change in the splicing pattern of the endogenous transformer and doublesex pre-mRNA of XX females from the female to the male mode. Consequently, these XX females were transformed into pseudomales. The comparison of the eleven Anastrepha Transformer-2 proteins among themselves, and with the Transformer-2 proteins of other insects, suggests the existence of negative selection acting at the protein level to maintain Transformer-2 structural features. Conclusions: These results indicate that transformer-2 is required for sex determination in Anastrepha through its participation in the female-specific splicing of transformer and doublesex pre-mRNAs. It is therefore needed for the auto-regulation of the gene transformer. Thus, the transformer/transfomer-2 > doublesex elements at the bottom of the cascade, and their relationships, probably represent the ancestral state ( which still exists in the Tephritidae, Calliphoridae and Muscidae lineages) of the extant cascade found in the Drosophilidae lineage ( in which tra is just another component of the sex determination gene cascade regulated by Sex-lethal). In the phylogenetic lineage that gave rise to the drosophilids, evolution co-opted for Sex-lethal, modified it, and converted it into the key gene controlling sex determination.

Excretion of ammonia by Lucilia cuprina larvae suppresses immunity in sheep

Attempts to immunise sheep against natural infestations by Lucilia cuprina larvae have not been effective. Yet it is known that the larvae excrete the immunosuppressant ammonium bicarbonate. The effect of larval ammonium and nonionic ammonia on immunopathobiology was evaluated in 12 infested sheep. The concentration of ammonium in veins draining infested sites was measured in another group of four sheep. Mean jugular unionized ammonia concentration increased 3.5 to 5.6 times above pre-infested control levels. Mean venous ammonium concentrations draining infested sites were 13 times higher than pre-infested jugular or carotid levels. Increases in jugular ammonia concentrations correlated with increased number of larvae, area of infestation, earlier death, neutropenia, eosinopenia, lymphocytopenia, large declines in serum globulins and zinc, and large rises in toxic neutrophils. The high concentrations of toxic unionized ammonia in blood directly permanently damaged neutrophils and lymphocytes and depressed serum globulin production. The results show that the ammonium from the excreta of larvae of L. cuprina may be highly immunosupressive. (C) 1997 Elsevier Science B.V.

Dispersão larval pós-alimentar de Lucilia sericata (Diptera, Calliphoridae) em condições de laboratório

Os dípteros califorídeos são os primeiros indivíduos a compor a sucessão faunística de cadáveres em decomposição, auxiliando assim na estimativa do intervalo pós-morte. Estudos de dispersão pós-alimentar de suas larvas possuem relevância para investigações médico-criminais. Diante disto, uma arena circular, simulando o ambiente natural, foi montada em laboratório a fim de verificar-se a dispersão larval radial pós-alimentar de Lucilia sericata (Meigen, 1826), tentando assim estabelecer relações entre as variáveis peso, distância e profundidade de enterramento na arena. Os resultados demonstraram que 45% das pupas foram recuperadas a uma profundidade compreendida entre 6 cm e 8 cm e a uma distância entre 33 cm e 45 cm do centro da arena. Não houve diferenças significativas quanto à propagação e profundidade de enterramento de machos, fêmeas e indivíduos inviáveis. Com relação ao peso, verificou-se que a média das fêmeas (x = 32,35 mg) foi superior a dos machos (x = 30,28 mg). A análise de correlação e de regressão entre peso e distância percorrida e entre peso e profundidade foram positivas, ou seja, pupas oriundas de larvas mais pesadas propagaram e se aprofundaram mais. O experimento permitiu ainda concluir que uma arena circular possibilita o deslocamento das larvas em todas as direções.

Estudio de cultivos celulares primarios derivados de Lucilia sericata (Diptera: 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.

Resistance to antiparasitic drugs: The role of molecular diagnosis

Chemotherapy is central to the control of many parasite infections of both medical and veterinary importance. However, control has been compromised by the emergence of drug resistance in several important parasite species. Such parasites cover a broad phylogenetic range and include protozoa, helminths and arthropods. In order to achieve effective parasite control in the future, the recognition and diagnosis of resistance will be crucial. This demand for early, accurate diagnosis of resistance to specific drugs in different parasite species can potentially be met by modern molecular techniques. This paper summarises the resistance status of a range of important parasites and reviews the available molecular techniques for resistance diagnosis. Opportunities for applying successes in some species to other species where resistance is less well understood are explored. The practical application of molecular techniques and the impact of the technology on improving parasite control are discussed. (C) 2002 Australian Society for Parasitology Inc. Published by Elsevier Science Ltd. All rights reserved.

Sobrevivência de pupas de Lucilia eximia (Wiedemann) (Diptera, Calliphoridae) após submersão em laboratório

Entre Maio e Junho de 2008 foi observado a sobrevivência das pupas de Lucilia eximia (Diptera, Calliphoridae) após submersão em laboratório. Para este experimento foram utilizadas 480 pupas de mesma idade, sendo estas divididas em oito grupos: o controle não foi submerso e os demais grupos foram submersos de um a sete dias. O aumento do período de submersão diminui a sobrevivência, com um dia de submersão a sobrevivência é de 80%, com dois dias 40%, é de 30% a partir do terceiro dia, no quarto dia cai para 23,34% e no quinto dia fica em 10%. Após este período a mortalidade sobe para 100%. Este padrão pode ser explicado pela curva "U-shaped" que ocorre no consumo de O2 durante o período pupal, onde o consumo é maior no início e no final do período pupal. O tempo de submersão também afeta o tempo de desenvolvimento, aumentando o período pupal. Estes dados têm o potencial para serem utilizados em investigações envolvendo Entomologia Forense, para a estimativa do tempo de submersão de um cadáver.

Diversity and Synanthropy of Calliphoridae (Diptera) in the Region of Rio Claro, SP, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Deep sequencing of New World screw-worm transcripts to discover genes involved in insecticide resistance

Background: The New World screw-worm (NWS), Cochliomyia hominivorax, is one of the most important myiasis-causing flies, causing severe losses to the livestock industry. In its current geographical distribution, this species has been controlled by the application of insecticides, mainly organophosphate (OP) compounds, but a number of lineages have been identified that are resistant to such chemicals. Despite its economic importance, only limited genetic information is available for the NWS. Here, as a part of an effort to characterize the C. hominivorax genome and identify putative genes involved in insecticide resistance, we sampled its transcriptome by deep sequencing of polyadenylated transcripts using the 454 sequencing technology. Results: Deep sequencing on the 454 platform of three normalized libraries (larval, adult male and adult female) generated a total of 548,940 reads. Eighteen candidate genes coding for three metabolic detoxification enzyme families, cytochrome P450 monooxygenases, glutathione S transferases and carboxyl/cholinesterases were selected and gene expression levels were measured using quantitative real-time polymerase chain reaction (qRT-PCR). Of the investigated candidates, only one gene was expressed differently between control and resistant larvae with, at least, a 10-fold down-regulation in the resistant larvae. The presence of mutations in the acetylcholinesterase (target site) and carboxylesterase E3 genes was investigated and all of the resistant flies presented E3 mutations previously associated with insecticide resistance. Conclusions: Here, we provided the largest database of NWS expressed sequence tags that is an important resource, not only for further studies on the molecular basis of the OP resistance in NWS fly, but also for functional and comparative studies among Calliphoridae flies. Among our candidates, only one gene was found differentially expressed in resistant individuals, and its role on insecticide resistance should be further investigated. Furthermore, the absence of mutations in the OP target site and the high frequency of mutant carboxylesterase E3 indicate that metabolic resistance mechanisms have evolved predominantly in this species.