Margareth Maria de Carvalho Queiroz

Morphological aspects of the larval instars of Chrysomya albiceps (Diptera, Calliphoridae) reared in the laboratory

In order to study the morphology of young Chrysomya albiceps forms, newly hatched larvae were collected at 2 hr intervals, during the first 56 hr; after this time the collection was made at 12 hr intervals. For identification and drawing, larvae were placed between a slide and a coverslip. The cephalopharyngeal skeletons along with the first and last segments were cut off for observation of their structures and spiracles. The larvae present microspines, which are distributed randomly throughout the 12 segments of the body surface; the cephalopharyngeal skeleton varies in shape and extent of sclerotization according to larval instar; the second and third instars have relatively long processes (tubercles) on the dorsal, lateral and ventral surfaces, with microspine circles on the terminal portion. Guimaraes et al. (1978) were the first researchers who recorded the presence of Chrysomya albiceps (Wiedemann, 1819), Chrysomya megacephala (Fabricius, 1974) and Chrysomya putoria (Wiedemann, 1818) in Brazil. According to them, these species were probably introduced in the southern region of Brazil by ships coming from Angola, Africa. These Diptera are considered important from the sanitary standpoint, since they may be mechanical vector of enteropathogens for humans as well as cause facultative myiasis. Today, they are widely distributed throughout the neotropical region (Prado & Guimaraes 1982, Baumgartner 1988), and have drawn the attention of several researchers. However, only the last two species have been well studied in terms of biology and taxonomy of the adults. The taxonomy of C. albiceps is still quite unknown since it was long thought to be indistinguishable from those of C. rufifacies (Macquart, 1842) (Patton 1922, Zumpt 1965, Kurahashi 1981, Richard & Gerrish 1983), a species that has not yet been recorded in Brazil. However, more accurate observations have shown some morphological aspects of the third instar of both species that allow their differentiation (Erzinclioglu 1987, Tantawi & Greenberg 1993). During the 70’s the two species were geographically isolated, C. albiceps being restricted to Africa and C. rufifacies to regions of Eastern Asia and Australia, although occupying the same biological niches in their respective natural biotopes

New records of calyptrate dipterans (Fanniidae, Muscidae and Sarcophagidae) associated with the decomposition of domestic pigs in Brazil.

The calyptrate dipterans are the most important decomposers of human cadavers. Knowledge of their species and distribution are of great importance to forensic entomology, especially because of the enormous diversity in Brazil. Carcasses of domestic pigs (Sus scrofa, L) were the experimental models used to attract calyptrates of forensic interest during the winters of 2006 and 2007 and the summers of 2006 and 2008. A total of 24,423 specimens from 44 species were collected (19 Muscidae, 2 Fanniidae and 23 Sarcophagidae), three of which were new records of occurrence and 20 of which were new forensic records for the state of Rio de Janeiro. Fourteen of these species were newly identified as forensically important in Brazil.

Population fluctuations of calliphorid species (Diptera, Calliphoridae) in the Biological Reserve of Tinguá, state of Rio de Janeiro, Brazil

The purpose of this work was to determine the diversity and population fluctuations of calliphorid flies in the Biological Reserve of Tingua (ReBio-Tingua), Nova Iguacu, state of Rio de Janeiro, Brazil and to correlate their occurrence with the environmental variables of temperature, rainfall and relative air humidity. Specimens of Diptera were collected monthly between June 2002 and January 2005 using four traps placed at four points along a trail and exposed for 48 hours. The traps were baited with sardines and the trapped insects were stored in 70% alcohol. It was collected 8,528 calliphorids, thirteen species were identified among the blowflies including Laneela nigripes Guimaraes 1977, Chrysomya megacephala (Fabricius, 1794), C. albiceps (Wiedemann, 1819), C. putoria (Wiedemann, 1830), Chloroprocta idioidea (Robineau-Devoidy, 1830), Cochliomyia macellaria (Fabricius, 1775), Hemilucilia semidiaphana (Rondani, 1850), H. segmentaria (Fabricius, 1805), Lucilia eximia (Wiedemann,1819), L. cuprina (Wiedemann, 1830), Paralucilia pseudolyrcea (Mello, 1969), Mesembrinella sp. and Eumesembrinella pauciseta (Aldrich, 1922). No significant correlation was found between the abundance of blowflies and the temperature and relative air humidity. Only C. megacephala and C. albiceps showed a positive and significant correlation with rainfall. An analysis of grouping by month (UPGMA) revealed no seasonal difference in the composition of the community, indicating that the community of calliphorid flies is probably more influenced by the ecological niches occupied by each species than by the seasons of the year.

Identification of fly eggs using scanning electron microscopy for forensic investigations

Forensic entomology is the science that studies the role of insects in decomposing corpses and one of the most common uses is to estimate the post-mortem interval (PMI) based on insect activity on a decomposing body. Usually, flies are the first insects to reach a carcass and are able to oviposit on carrion within a few hours after death. Scanning electron microscopy (SEM) gives detailed information about morphological characters helping to identify the immature forms of flies and consequently serves as a tool in crime scene investigations. Sometimes, only eggs and larvae are found in corpses. Some dipteral species are important because their larvae develop in organic matter. The aim of this study is to identify eggs of species of forensic importance, such as Chrysomya megacephala, Chrysomya putoria, Lucilia cuprina, Lucilia eximia and Ophyra aenescens, using scanning electron microscopy (SEM). C. megacephala had no anastomosis or holes at the top of the islands and C. putoria had few anastomoses and no holes, whereas L. eximia and O. aenescens were found to have anastomoses and holes and L. cuprina had only anastomoses. The median area was bifurcated anteriorly in C. megacephala, L. eximia and O. aenescens and rounded in C. putoria and L. cuprina. Also the sculptures observed in the chorionic cells, the length and the way that median area ends up posteriorly are characteristics of great diagnostic value to identify muscoids of forensic importance.

Diversity of Calliphoridae (Diptera) in Brazil's Tinguá Biological Reserve

The Calliphoridae are flies of great ecological, medical and sanitary importance because they are decomposers of organic matter, mechanical vectors of pathogenic agents, and causers of myiasis. This paper attempts to ascertain the diversity of Calliphoridae in the Tinguá Biological Reserve and correlate meteorological data (temperature, relative humidity of the air, and precipitation) with the occurrence of these flies. The study was conducted at a site in the Tinguá Biological Reserve, located in the municipality of Nova Iguaçu, state of Rio de Janeiro, Brazil. Four traps were set up using chicken viscera as bait. The experiment was conducted over the period of May 2001 to April 2002, with two monthly collections. The captured flies were killed with ether and conserved in 70% alcohol and identified in the Laboratory of Diptera Studies (UNIRIO) and the Laboratory of Vector Insect Biology and Control (FIOCRUZ), both headquartered in the city of Rio de Janeiro, RJ, Brazil. A total of 1,987 insects were captured, 37.5% belonging to the family Calliphoridae, 29.4% to Muscidae, 16.0% to Sarcophagidae, and 17.1% to other families. The most representative species found was Phaenicia eximia (47.0%), followed by Hemilucilia semidiaphana (23.6%), Mesembrinella bellardiana (13.7%), Hemilucilia segmentaria (7.5%), Chloroprocta idioidea (4.9%), Chrysomya albiceps (1.2%), Chrysomya megacephala (0.9%), Phaenicia sericata (0.6%), Eumesembrinella sp. (0.5%), and Chrysomya putoria (0.1%). Large numbers of Calliphoridae were noted in May, June, September and January, coinciding with low rainfall and an average temperature of 21.8 to 27.0 degrees C. A negative correlation was found between the number of Calliphoridae captured and meteorological factors.

Temperature requirements of Chrysomya albiceps (Wiedemann, 1819) (Diptera, Calliphoridae) under laboratory conditions

Chrysomya albiceps specimens were obtained from colonies established with larvae and adults collected at the Federal Rural University in Rio de Janeiro, Seropédica, State of Rio de Janeiro. The larval stage of C. albiceps was allowed to develop in climatic chambers at temperatures of 18, 22, 27 and 32 degrees C, and the pupal stage was allowed to develop at 22, 27 and 32 degrees C (60 +/- 10% RH and 14 hr photoperiod). The duration and viability of the larval stage of C. albiceps at 18, 22, 27 and 32 degrees C were 21.30, 10.61, 5.0 and 4.0 days and 76.5, 88.5, 98.5 and 99.5%, respectively, with mean mature larval weights of 45.16, 81.86, 84.35 and 70.53 mg, respectively. Mean duration and viability of the pupal stage at 22, 27 and 32 degrees C were 9.36, 4.7 and 3.0 days and 93.8, 100 and 100%, respectively. The basal temperature for the larval and pupal stage and for the larval and adult phase were 15.04, 17.39 and 15.38 degrees C, corresponding to 65.67, 44.15 and 114.23 DD.

Ultramorphological characteristics of immature stages of Chrysomya albiceps (Wiedemann 1819) (Diptera: Calliphoridae), a fly specie of forensic importance.

Forensic entomology is an area of science that serves as a tool in crime scene investigations. Usually, flies are the first insects to reach a dead body and can oviposit just a few hours after arrival. Therefore the knowledge of immature stages is essential for correct identification of the species found on corpses. Scanning electron microscopy gives detailed information about morphological characters helping to identify the immature forms and consequently serves as a tool in crime scene investigations. C. albiceps is a very important fly for forensic entomologists because its larvae are almost always present on a dead body and it is facultative predators and therefore can alter the composition of species present at the carcass. The aim of this study is to identify eggs, larvae, and puparia of C. albiceps using SEM. Eggs were elongated with the anterior region ending in a “Y” shape and the posterior end was tapered. The micropyle was a well‐adorned orifice with some projections around it. The first instar larva was composed of 12 segments separated by spines. Only one spiracular opening could be seen at the posterior spiracle. Body tegument was smooth and tubercles were not seen. Antennae and maxillary palps were visible. Second and third larval instars were very similar to first instar, except for the presence of anterior spiracle. However, body tegument was composed of net‐like patches and tubercles were visible. Tubercles present at the third instar larvae were robust and erect. Puparia showed a retracted cephalic region and curved tubercles. Microsc. Res. Tech. 73:779–784, 2010.

Factors of susceptibility of human myiasis caused by the New World screw-worm, Cochliomyia hominivorax in São Gonçalo, Rio de Janeiro, Brazil

Abstract This study was carried out between July 2007 and June 2008 and reports on the occurrence of human myiasis caused by the New World screwworm, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae) in São Gonçalo in the state of Rio de Janeiro, Brazil. Liquid or solid vaseline was used to suffocate the larvae, which were then preserved in 70% ethanol and sent to the Instituto Oswaldo Cruz for identification. C. hominivorax were identified in all 22 cases of myiasis. There were 12 male and 10 female patients with ages ranging from 03 to 71. Ethnically the highest incidence was among black people, with 17 cases. Open wounds were the main cause of the parasitosis, whereas poor personal hygiene, the low educational level, alcoholism, bedridden patients, and physical or mental disability were possibly secondary factors; in addition to all these factors the income of the patients was very low.

Cuticular hydrocarbons as a tool for the identification of insect species: puparial cases from Sarcophagidae.

The external surface of all insects is covered by a species-specific complex mixture of highly stable, very long chain cuticular hydrocarbons (CHCs). Gas chromatography coupled to mass spectrometry was used to identify CHCs from four species of Sarcophagidae, Peckia (Peckia) chrysostoma, Peckia (Pattonella) intermutans, Sarcophaga (Liopygia) ruficornis and Sarcodexia lambens. The identified CHCs were mostly a mixture of n-alkanes, monomethylalkanes and dimethylalkanes with linear chain lengths varying from 23 to 33 carbons. Only two alkenes were found in all four species. S. lambens had a composition of CHCs with linear chain lengths varying from C23 to C33, while the other three species linear chain lengths from 24 to 31 carbons. n-Heptacosane, n-nonacosane and 3-methylnonacosane, n-triacontane and n-hentriacontane occurred in all four species. The results show that these hydrocarbon profiles may be used for the taxonomic differentiation of insect species and are a useful additional tool for taxonomic classification, especially when only parts of the insect specimen are available.

Effects of latex from “Amapazeiro” Parahancornia amapa (Apocynaceae) on blowfly Chrysomya megacephala (Diptera: Calliphoridae) post-embryonic development

Nowadays, insect control is usually carried out using chemical insecticides, but insect resistance and other negative side effects have prompted the search for alternatives. Biopesticides provide a positive alternative to synthetic pesticides because they have low impact on the environmental, low toxicity to humans and low costs among other advantages. This research was carried out to evaluate the activity of Parahancornia amapa (Huber) Ducke (Apocynaceae) lyophilized latex on the post embryonic development of Chrysomya megacephala (F.) (Diptera: Calliphoridae). Larvae treated with 1.0% latex showed a shorter post embryonic development period (larval, pupal and newly hatched larvae to adult); whereas larvae treated with 3.0% latex provoked a prolongation of these periods. Viability (53%) was also very low at the newly hatched larvae to adult period for larvae treated with 3.0% latex, indicating that latex from P. amapa at high concentrations could change C. megacephala post embryonic development.