Selection And Validation Of Reference Genes For Functional Studies In The Calliphoridae Family.

The genera Cochliomyia and Chrysomya contain both obligate and saprophagous flies, which allows the comparison of different feeding habits between closely related species. Among the different strategies for comparing these habits is the use of qPCR to investigate the expression levels of candidate genes involved in feeding behavior. To ensure an accurate measure of the levels of gene expression, it is necessary to normalize the amount of the target gene with the amount of a reference gene having a stable expression across the compared species. Since there is no universal gene that can be used as a reference in functional studies, candidate genes for qPCR data normalization were selected and validated in three Calliphoridae (Diptera) species, Cochliomyia hominivorax Coquerel, Cochliomyia macellaria Fabricius, and Chrysomya albiceps Wiedemann . The expression stability of six genes ( Actin, Gapdh, Rp49, Rps17, α -tubulin, and GstD1) was evaluated among species within the same life stage and between life stages within each species. The expression levels of Actin, Gapdh, and Rp49 were the most stable among the selected genes. These genes can be used as reliable reference genes for functional studies in Calliphoridae using similar experimental settings.

Comunidades de formigas (Hymenoptera, Formicidae) em fragmentos de Mata Atlântica situados em áreas urbanizadas

As comunidades de formigas que vivem em fragmentos de Mata Atlântica rodeados por um ecossistema urbano bem desenvolvido, foram investigadas. Avaliou-se a riqueza, a freqüência de ocorrência das espécies bem como a similaridade entre três áreas da cidade de São Paulo: Parque da Previdência (PP), Reserva Florestal "Armando Salles de Oliveira" (CUASO) e Horto Oswaldo Cruz (HOC). Foram colocadas armadilhas do tipo "pitfall" em locais onde não ocorre visitação pública, durante uma semana, nos meses de março, junho, setembro e dezembro de 2001. Em todos os fragmentos foram coletadas 79 espécies de formigas, pertencentes a nove subfamílias e 32 gêneros. A subfamília Myrmicinae e os gêneros Pheidole e Hypoponera foram os mais ricos. No PP foram registradas 62 espécies, na CUASO 46 e no HOC 43, sendo que PP e CUASO são mais similares entre si. Tal similaridade possivelmente esteja relacionada ao tamanho de ambas as áreas e, também, a uma semelhança nos sítios de nidificação e de alimentação. No geral, a fauna de formigas é generalista, com a presença de alguns gêneros especialistas, como Discothyrea, Acanthognathus, Gnamptogenys, Oxyepoecus e Pyramica; ou de gêneros cujos hábitos alimentares ainda são desconhecidos (Heteroponera e Myrmelachista). A presença de espécies caracteristicamente de áreas domiciliares também foi constatada: Pheidole megacephala Fabricius, 1793, Linepithema humile Mayr, 1868, Wasmannia auropunctata Roger, 1863, Paratrechina fulva Mayr, 1862, P. longicornis Latreille, 1802 e Tapinoma melanocephalum Fabricius, 1793.

The influence of temperature and humidity on abundance and richness of Calliphoridae (Diptera)

The blowfly species are important components in necrophagous communities of the Neotropics. Besides being involved in the degradation of animal organic matter, they may serve as vectors for pathogens and parasites, and also cause primary and secondary myiasis. The occurrence pattern of these species is well defined, yet it is still not very clear which of these environmental factors determine the structure of the assemblies. This paper was developed to evaluate the influence of mean temperature and relative humidity variation in the abundance and richness of blowflies in the Brazilian southernmost state, Rio Grande do Sul, where temperature variation is well marked throughout the year. To evaluate this objective, WOT (Wind Oriented Trap) were installed with beef liver as bait in three environments for 10 consecutive days in each month between July 2003 and June 2004. A total of 13,860 flies were collected distributed among 16 species with a higher frequency of Lucilia eximia (Wiedemann, 1819) and Chrysomya albiceps (Wiedemann, 1819). The mean temperature and relative humidity influence the richness of blowflies, with greater richness and abundance in late spring and early summer, whereas abundance was only influenced by temperature. Each species responded differently with respect to these variables, where L. eximia is not influenced by any of the two abiotic factors, despite the high abundance presented. This paper presents the results of the sensitivity for the presence or absence of species of Calliphoridae and on the variation of the abundance of these species under regime temperature changes and relative humidity with implications for public health and animal management.

Substratos utilizados para a criação de dípteros caliptratos em uma área urbana do Município do Rio de Janeiro

Em uma área urbana do município do Rio de Janeiro, foram criados 10.988 dípteros caliptratos, pertencentes a quatro famílias e 22 espécies. As criações foram efetuadas em quatro etapas durante os anos de 1986 a 1987. Foram utilizados como substratos: peixe, fígado, carcaça de camundongo, fezes humanas e caninas, camarão, siri, banana, manga mamaão e tomate. As espécies criadas com maior freqüência foram: Fania sp (subgrupo pusio), Atherigona orientalis, Phaenicia eximia, Paraphrissopoda chrisostoma, Chrisomyia megacephala, Ophyra solitaria, Musca domestica, Synthesiomyia nudiseta, Phaenicia cuprina, Ophyra aenescens, Sarcophagula occidua, Morellia flavicornis, e Sarcodexia innota.

Substratos utilizados para a criação de dípteros caliptratos no jardim zoológico do Rio de Janeiro (Rio-Zoo)

No Rio-Zoo, foram criados 7.377 dípteros caliptratos, pertencentes a quatro famílias e 22 espécies. As criações foram efetuadas de agosto de 1987 a abril de 1988. Foram utilizados como substratos de criação: banana, camarão, carcaça de camundongo,fezes caninas, fezes humanas, fígado bovino, lula, mamão, peixe, siri e tomate. As espécies criadas com maior freqüência foram: Fannia sp. (subgrupo pusio), Atherigona orientalis, Chrysomyia megacephala, Phaenicia eximia. Paraphrissopoda chrysosotoma, Ophyra aenescens, Synthesiomyia nudiseta, Ophyra chalcogaster, Oxisarcodexia fluminensis e Hemilucilia segmentaria. Foi efetuado um estudo, ainda que incompleto, das espécies que se desenvolvem em fezes de animais cativos do Rio-Zoo. Dentre as mais freqüentes destacam-se: Fannia sp., Sarcophagula occidua, Ophyra chalgogaster, Ravinia belforti e Phaenicia eximia.

Diseminación de enteroparasitos por Calliphoridae (Insecta, Diptera)

It is establish the dissemination enteroparasite by Calliphoridae in a district situated around by Federal Capital of Argentina. The species implicated in this dispersal are: Phaenicia sericata (meigen, 1826); Calliphora vicina Robineau-Desvoidy, 1830; Chrysomya chloropyga (Wiedemann, 1818) and Phaenicia eximia (Wiedemann, 1819). Fifty two flies was studies, thirty four (65%) to belong at the family Calliphoridae. Of this thirteen (38%) have eggs of taeniid and cysts of Entamoeba coli (Grassi, 1879) and Giardia lamblia Styles, 1915.

Levantamento de microhimenópteros parasitóides de dípteros de importância, médico-veterinária no Brasil

De larvas e pupas de Musca domestica, Chrysomya albiceps, Cochliomyia homivorax, Stomoxys calcitrans e Syntesiomyia nudiseta coletadas em diversos ambiente, em São Paulo, Paraná, Mato Grosso, Mato Grosso do Sul e Minas Gerais, foram obtidas dez espécies de microhimenópteros parasitóides da supermamília Chalcidoidea, algumas assinaladas pela primeira vez no Brasil.

A checklist of arthropods associated with pig carrion and human corpses in Southeastern Brazil

Necrophagous insects, mainly Diptera and Coleoptera, are attracted to specific stages of carcass decomposition, in a process of faunistic succession. They are very important in estimating the postmortem interval, the time interval between the death and the discovery of the body. In studies done with pig carcasses exposed to natural conditions in an urban forest (Santa Genebra Reservation), located in Campinas, State of São Paulo, southeastern Brazil, 4 out of 36 families of insects collected - Calliphoridae, Sarcophagidae, Muscidae (Diptera) and Dermestidae (Coleoptera) - were considered of forensic importance, because several species were collected in large numbers both visiting and breeding in pig carcasses. Several species were also observed and collected on human corpses at the Institute of Legal Medicine. The species belonged to 17 different families, 6 being of forensic importance because they were reared from human corpses or pig carcasses: Calliphoridae, Sarcophagidae, Muscidae, Piophilidae (Diptera), Dermestidae, Silphidae and Cleridae (Coleoptera). The most important species were: Diptera - Chrysomya albiceps, Chrysomya putoria, Hemilucilia segmentaria, Hemilucilia semidiaphana (Calliphoridae), Pattonella intermutans (Sarcophagidae), Ophyra chalcogaster (Muscidae), Piophila casei (Piophilidae); Coleoptera - Dermestes maculatus (Dermestidae), Oxyletrum disciolle (Silphidae) and Necrobia rufipes (Cleridae).

Incidence of myiasis in Panama during the eradication of Cochliomyia hominivorax (Coquerel 1858, Diptera: Calliphoridae) (2002-2005)

We present the results of a study on myiasis in Panama during the first years of a Cochliomyia hominivorax eradication program (1998-2005), with the aim of investigating the behavior of the flies that produce myiasis in animals and human beings. The hosts that registered positive for myiasis were cattle (46.4%), dogs (15.3%), humans (14.7%), birds (12%), pigs (6%), horses (4%), and sheep (1%). Six fly species caused myiasis: Dermatobia hominis (58%), Phaenicia spp. (20%), Cochliomyia macellaria (19%), Chrysomya rufifacies (0.4%), and maggots of unidentified species belonging to the Sarcophagidae (3%) and Muscidae (0.3%). With the Dubois index, was no evidence that the absence of C. hominivorax allowed an increase in the cases of facultative myiasis.

Novos táxons de Hemilophini (Coleoptera, Cerambycidae, Lamiinae) sem carenas nos élitros da Região Neotropical

Novos registros e novas espécies de Hemilophini (Cerambycidae) coletados por pulverização de inseticida nas copas das árvores no Equador: Olivensa megacephala (Bates, 1866); Icupima laevipennis (Gahan, 1892); Hemilomecopterus alienus Galileo & Martins, 2004; Acapiata gen. nov., espécie-tipo A. dilatata sp. nov.; Acaiu gen. nov., espécie-tipo A. spinosus sp. nov.; Okamira gen. nov., espécie-tipo O. pulchra sp. nov. Duas espécies novas de Erana Bates, 1866 também são descritas: E. costaricensis sp. nov. da Costa Rica (Heredia) e E. flaviventris sp. nov. de Honduras (Yoro).

Insects of forensic importance from Rio Grande do Sul state in southern Brazil

The experiment was conducted throughout the year 2005, at the Universidade Federal de Pelotas campus. The objectives of the study were to analyze the decomposition of rabbit (Oryctolagus cunniculus L.) with mean weight 2.67 Kg carcass and describe the interaction of insects acting on it, as well as the insect's potential use in legal medicine. We collected 5.239 insect specimens; 1.827 of them were obtained from larvae collected from carcasses and reared. The specimens were identified and 20 species were of forensic importance. The species Lucilia eximia (Wiedemann, 1819) and Chrysomya albiceps (Wiedemann, 1819) (Diptera, Calliphoridae) were better indicators of post-mortem interval (PMI) because they occurred in all seasons and were the first to reach the carcass. Hemilucilia semidiaphana (Rondani, 1850), H. segmentaria (Fabricius, 1805) (Diptera, Calliphoridae), Muscina stabulans (Fallén, 1817) and Synthesiomyia nudiseta (Wulp, 1883) (Diptera, Muscidae) can disclose death time because they occur only in certain months of the year. Oxyletrum discicolle (Brullé, 1840) (Coleoptera, Silphidae) and Dermestes maculates De Geer, 1774 (Coleoptera, Desmestidae) were found in advanced stages of decomposition.

Rate of development of forensically-important Dipterain southern Brazil

Dipteran larvae were collected from rabbit (Oryctolagus cunniculus L.) carcasses during the four seasons in 2005 in the southernmost state of Rio Grande do Sul, Brazil. The larvae were fed ground beef at ambient temperatures following collection from carcasses. The development of each species under these conditions was estimated. The most abundant species in the carcasses were Lucilia eximia (Wiedemann) and Chrysomya albiceps (Wiedemann) (Calliphoridae), and they were found in all seasons. The data were fitted to a linear model that describes the relationship between temperature and linear developmental rating. These two species are primary forensic indicators in southern Brazil. Other species such as Hemilucilia semidiaphana (Rondani) (Calliphoridae), Synthesiomyia nudiseta (Wulp), Muscina stabulans (Fallen) (Muscidae), and Fannia pusio (Wiedemann) (Fanniidae) were forensically less important because they only occurred in high frequency in certain seasons and during the first days of carcass decomposition.

Insects associated with exposed decomposing bodies in the Colombian Andean Coffee Region

In Colombia, mainly classic forensic medicine methods were used to clarify crimes until 2004. However, other disciplines, including forensic entomology, started to be considered only after the New Accusatory System introduction in Bogotá and the Coffee Region in 2005. In order to provide tools for obtaining evidentiary material elements in judicial trials, it is presented here the succession of insects throughout the decomposition process of an exposed carcass of Sus scrofa Linnaeus 1758 (Suidae) and the Occurrence Matrix of colonizing species. This process was evaluated under ambient conditions in the Andean rural area of the city of Pereira, in the Mundo Nuevo district, located in a pre-montane Wet Forest area, from October to November 2006. A sampling period of 27 days and 3198 individuals were collected. We found these colonizing species in the following stages of decomposition: Lucilia eximia (Wiedemann, 1819) fresh; Hemilucilia semidiaphana (Rondani, 1850), Oxelytrum discicolle (Brullé, 1840), and Cochliomyia macellaria (Fabricius 1775) bloated; Chrysomya albiceps (Wiedemann 1819), Compsomyiops verena (Walker, 1849), Ophyra aenescens (Wiedemann, 1830) and Musca domestica Linnaeus, 1758 active; Fannia sp. advanced and Stearibia nigriceps (Meigen, 1826) remains. This study provides support tools to define the Post Mortem Interval that may be used by experts from government institutions and laboratories officially accredited.

Preliminary study of insects associated to indoor body decay in Colombia

Preliminary study of insects associated to indoor body decay in Colombia. This is the first report studying insects associated to indoor body decay process of a white pig (Sus scrofa) (Artiodactyla, Suidae) in a controlled indoor environment in an urban area of Florencia city, Amazonia Piedmont, Colombia. For a period of 54 days, 9,220 individuals (immature and adults), distributed in 3 orders, 5 families, 10 genera, and 10 species were collected using entomological nets and tweezers. Five decaying stages are described (fresh, bloated, active decay, advanced decay and remains). During the fresh stage we recorded Cochliomyia macellaria (Fabricius, 1775), Chrysomya albiceps (Wiedemann, 1819), Ophyra aenescens (Wiedemann, 1830), Oxysarcodexia sp., Lepidodexia sp. and Lasiophanes sp.; during the bloating stage C. macellaria, C. albiceps, Lucilia eximia (Wiedemann, 1819), Hemilucillia semidiaphana (Rondani, 1850), Musca domestica Linnaeus, 1758, O. aenescens, Oxysarcodexia sp., Lepidodexia sp., Dermestes maculatus De Geer, 1774 and Lasiphanes sp.; during the active decay C. macellaria, C. albiceps, L. eximia, M. domestica, O. aenescens, Lepidodexia sp. D. maculatus and Lasiophanes sp.; during the advanced decay C. macellaria, C. albiceps, M. domestica, Lepidodexia sp. and Lasiophanes sp.; and during the remains stage C. albiceps, D. maculatus and Lasiophanes sp. The insects were sorted out in 3 ecological categories; necrophagous, predators and parasites and sarco-saprophagous. According to Chao and Jack estimators, total richness was observed on day 20, with 100% of the expected species.

Necrophagous diptera associated with wild animal carcasses in southern Brazil

Necrophagous Diptera associated with wild animal carcasses in southern Brazil. The aim of this study was to acquire a better knowledge concerning the diversity of necrophagous Diptera that develop on wild animal carcasses. For this purpose, the decomposition of six wild animal carcasses was observed in order to collect and identify the main species of necrophagous flies associated with the decomposition process. The carcasses were found on highways near the cities of Pelotas and Capão do Leão in the initial stage of decomposition, with no significant injuries or prior larval activity. Four wild animal models were represented in this study: two specimens of Didelphis albiventris Lund, 1840; two Tupinambis merianae Linnaeus, 1758; one Nothura maculosa Temminck, 1815; and one Cerdocyon thous Linnaeus, 1766. A total of 16,242 flies from 14 species were reared in the laboratory, where Muscidae presented the greatest diversity of necrophagous species. Overall, (i) carcasses with larger biomass developed a higher abundance of flies and (ii) the necrophagous community was dominated by Calliphoridae, two patterns that were predicted from published literature; and (iii) the highest diversity was observed on the smaller carcasses exposed to the lowest temperatures, a pattern that may have been caused by the absence of the generalist predator Chrysomya albiceps (Wiedemann, 1819). (iv) An UPGMA analysis revealed a similar pattern of clusters of fly communities, where the same species were structuring the groupings.