First record of Brachymeria amenocles (Hymenoptera: Chalcididae) associated with larvae of Peckia (Peckia) chrysostoma (Diptera: Sarcophagidae) in Brazil.

In this work, we report for the first time the occurrence of the parasitoid wasp Brachymeria amenocles (Walker, 1846) (Hymenoptera: Chalcididae) associated with the larvae of the flesh fly, Peckia (Peckia) chrysostoma (Wiedemann, 1830) (Diptera: Sarcophagidae), in Brazil. This parasitoid species was collected together with the species Brachymeria podagrica (Fabricius, 1787), in the municipality of Rio de Janeiro, geographically located in the Southeast region of Brazil, in larvae of the same host species reared in the same attraction substrate, behaving as solitary parasitoids. A trap containing putrefying sardine was exposed for 48 h, in the upper part of a house, located in Vila Isabel, an urban area. The larvae collected were reared in the laboratory without control of environmental conditions. A total of three adult parasitoid wasps of the species B. podagrica, and nine adult parasitoid wasps of the species B. amenocles emerged from 12 host pupae, resulting in a prevalence of parasitoidism of 4.8% and 14.5%, respectively. Developing parasitoids at the pupal phase were observed in another 34 host pupae dissected.

Biological behavior of Chrysomya putoria (Wiedemann, 1819) (Diptera: Calliphoridae) after refrigeration: Logistics for use in Biotherapy.

The influence of refrigeration on the post-embryonic development of Chrysomya putoria larvae was evaluated, regarding its resistance in the logistics of storage and distribution in biotherapy. Previously sterilized larvae were submitted to four periods of storage under refrigeration (T1=12 h, T2=24 h, T3=48 h and T4=72 h) and control (without sterilization and refrigeration). Newly hatched larvae (0.200 g) were stored between 3 and 9ºC. After refrigeration, 40 neo-larvae (in triplicate) were transferred to 50 g of protein diet and incubated in an acclimatized chamber. There was a significant difference in the larval body mass (T1 and T2) and in the duration of larval, pupal and total development (T3 and T4). The sex ratios found in the four treatments did not differ from what was expected. Normality rates were 100% for all treatments. There was no significant difference between the Control, T1 and T2 treatments for larval, pupal and total viability. There was a significant difference between control (C) and T4 (larval viability), between C, T3 and T4 (pupa) and between C and T4 (total). C. putoria has resistance under refrigeration and storage of up to 56 h, presenting viability above 70% for use in biotherapy.

Post-embriononic development of Chrysomya putoria(Diptera: Calliphoridae) on a diet containing ampicillin in different concentrations.

Here we evaluate the effects of different concentrations of the antibiotic ampicillin on the growth and development of Chrysomya putoria. Third-generation, first instar larvae (L1) reared on 60 grams of homogenate+agar 65% were treated with ampicillin sodium. The experiment consisted of four replicates (40 larvae/replicate) of each antibiotic concentration tested (T1: 466µg/mL ; T2: 81.33 mg/mL and T3: 166.66mg/mL) and a T4: control. The body mass of the mature larvae, after they abandoned the diet, were recorded in batches of five. The variation between the mean body mass of larvae and the duration of larval and pupal stages, and overall duration of the development, viability and normal rates were analyzed by ANOVA. There were no significant differences between the four treatments in the following parameters: body mass of larvae that discontinued the diet as well as the duration of larval, pupal, and total development. The sex ratios found in the four treatments did not differ from those expected. Normality rates were 100% for all treatments. There were no significant differences between treatments for larval and overall viability, but pupal viability differed significantly between T1 and the control, T1 and T2, and between the control and T3. The antibiotic did not appear to significantly alter the development of C. putoria.

Intrapuparial development of Chrysomya putoria (Diptera: Calliphoridae).

The intrapuparial development of 150 pupae of Chrysomya putoria (Wiedemann, 1830) was analyzed. The material was kept in a climate chamber at 27 degrees C (day) and 25 degrees C (night), 60 +/- 10% relative humidity (RH), and a photoperiod of 14:10 (L:D) h, which started at 06:00 a.m. Daily, 30 pupae were frozen at - 15 degrees C, for the sacrifice of the insects, and the immatures obtained were photomicrographed until the emergence of the adults. The following stages were observed: pupariation process: during this process, reduction of the larvae's first three body segments and the darkening of the cuticle occur; cryptocephalic phase after 18 h; phanerocephalic phase after 24 h; body division after 48 h. Between the third and the fourth days of the development, the stage ofpharate adult was reached, characterized by the following changes in the color of the compounds eyes: 1) orange eyes, after 66 h; 2) red eyes, after 72 h; 3) dark red eyes, after 90 h; 4) brown eyes, after 96 h. The time of intrapuparial development of C. putoria at 2 degrees C (day) and 25 degrees C (night), 60 +/- 10% RH, and a photoperiod of 14:10 (L:D) h was 5 d.

Effects of the antibiotics Gentamicin on the postembryonic development of Chrysomya putoria (Diptera: Calliphoridae).

We evaluate the effects the antibiotic Gentamicin on the development of Chrysomya putoria (Wiedemann, 1818). Third-generation, first-instar larvae were reared in a climatic chamber on 60 g of homogenate + agar 65% and were treated with three concentrations of Gentamicin: 4.44 mg/ml, 13.33 mg/ml, and 66.66 mg/ml. The control consisted of distilled water. The relationships between mean body mass of mature larvae (measured after diet abandonment, in batches of five individuals), duration of larval and pupal stages, and overall duration of development were analyzed. The actual sex ratio was compared against the expected using the chi square. None of the parameters measured differed significantly among the four treatments, with one exception: when Gentamicin concentration was 13.33 mg/ml, larval viability differed significantly from the control. All larvae from all treatments were considered normal. We conclude that the antibiotic did not significantly alter the development of C. putoria (Wiedemann) (Diptera: Calliphoridae).

Biological Response of Chrysomya putoria (Wiedemann, 1818) (Diptera: Calliphoridae) Pupae After Submersion in Freshwater.

Forensic Entomology uses arthropods to aid in legal investigations. This study checked the biological response of Chrysomya putoria pupae to submersion in fresh water for up to 6 d, evaluating the critical submersion time, survival rate, and development time of the flies. Adults were collected using fish baits in two typical traps. Seven hundred and twenty fourth-generation pupae with 2 d of development were used and separated into submergence intervals: 24, 48, 72, 96, 120, and 144 h. An additional 120 pupae were used as a control. Each treatment was done in triplicate, consisting of 40 pupae distributed in four tulle-sealed test tubes containing 10 pupae each. All tubes of each treatment were co-adhered in test tube racks and were submerged in mineral water in a container with constant oxygenation, except those of the control group, which were not submerged. The tubes were removed from the water according to their respective submersion interval, until 144 h was completed. The control group had a survival rate of 90%, while the 24-h treatment had 85% and the 48-h treatment had 35.8%. The critical submersion time for pupae was 72 h, with 100% mortality by 144 h. The average development time for the control group was 3.2 d, while the 24- and 48-h treatments developed in 4.3 and 6.3 d, respectively. The longer the individuals were submerged, the lower the survival rate was, while the development time increased. The data obtained in this study have potential in applications to estimate the interval of submersion of a cadaver.

Evaluation of Chilean Boldo Essential Oil as a Natural Insecticide Against Chrysomya megacephala (Diptera: Calliphoridae).

Chrysomya megacephala (Fabricius, 1794) is a vector of enteric bacteria, protozoa, helminths, and viruses. These Diptera can also be responsible for secondary myiasis in several animal species. Therefore, it is easy to understand the relevance of studies focusing on C. megacephala dissemination control. The employment of essential oils as natural insecticides must be considered as a promising alternative for the replacement of synthetic insecticides. In this context, the essential oil obtained from Chilean boldo leaves should be highlighted. The aim of the present work was to assess the insecticidal activity of Chilean boldo essential oil against C. megacephala blowflies in different life stages (larva, pupa, and adult). The essential oils were extracted from commercial samples of Chilean boldo leaves by hydrodistillation and were mixed to produce a pool that was employed in the study. Gas chromatographic techniques were used to enable the identification and quantification of the pool's components. Larvae, pupae, and adult insects of C. megacephala were exposed (topical application) to different concentrations of this essential oil pool. After that, the larvicidal, pupicidal, and insecticidal actions of the oil were tested. Its toxicity might be associated with compounds such as eucalyptol, linalool, α-pinene, limonene, and ascaridole, either acting alone or by synergic effects. Interestingly, the pupae appeared to be stronger than the larvae and adult insects, needing higher doses of essential oil to be killed. The oil's toxic effects could be useful to control C. megacephala dissemination in all of its development phases.

Population Fluctuation, Influence of Abiotic Factors and the Height of Traps on the Abundance and Richness of Calliphoridae and Mesembrinellidae.

Calliphoridae and Mesembrinellidae are relevant to environmental conservation, public health, and forensic entomology. Researches regarding the flight behavior and the influence of abiotic factors on these insects may assist the application of entomology sciences. This study aimed to analyze the population fluctuation of Calliphoridae and Mesembrinellidae, verifying the influence of environmental factors, trap height, and the anthropic effect in the Itaipu-Piratininga lagoon complex, Niterói (RJ). The collections were carried out monthly from September 2015 to August 2016, with fish bait (sardines) exposed for 48 h, totaling six traps, installed in three physiognomies (mangrove, ombrophilous forest, and restinga) at 1.5 and 2.5 m from the ground. Nine thousand seven hundred seventy-three individuals were captured, comprising two families, five genera and 11 species. Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) was the predominant species and Mesembrinella bellardiana (Aldrich, 1922) (Diptera: Mesembrinellidae) the least representative, indicating the low preservation level of this ecosystem. There was a weak, positive correlation between abundance of Chloroprocta idioidea (Robineau Desvoidy, 1830) (Diptera: Calliphoridae), Ch. megacephala, Chrysomya putoria (Wiedemann, 1818) (Diptera: Calliphoridae), and Lucilia cuprina (Wiedemann, 1830) (Diptera: Calliphoridae) with the temperature, as well as between the abundance of C. idioidea and Chrysomya albiceps (Wiedemann, 1819) (Diptera: Calliphoridae) with precipitation; however, there was no correlation between abundance and relative humidity. No significant influence of the trap height was observed. We stated a high influence of anthropic effects on the restinga and mangrove physiognomies, while the forest physiognomy still retains its preserved characteristics, with the dominance of forestall species.

Efficacy of UV-C Ray Sterilization of Calliphora vicina (Diptera: Calliphoridae) Eggs for Use in Maggot Debridement Therapy.

Maggot debridement therapy (MDT) is a simple wound debridement technique. It is a natural treatment licensed by the Food and Drug Administration (FDA) and is increasingly used in the United States and in Europe. This treatment is safe when the larvae originate from laboratory stocks of eggs that have been sterilized. In this study, a simple, inexpensive microbe decontamination technique is described. It yields eggs that are free of chemical residues and are easy to handle, meeting the growing demand for medicinal larvae in hospitals or medical centers. Three treatments (T1, T2, T3) involving 3, 6, and 12 min of exposure to ultraviolet (UV-C) rays, respectively, were compared. Egg sterility was evaluated by culture in thioglycollate broth, incubated at 32°C ± 2.5°C under aerobic conditions for up to 14 d. The UV-C radiation sterilization process obtained satisfactory results after 12 min exposure (treatment 3). Larval viability was 57%, pupal viability was 54%, and 54% of the adults emerged. The sex ratio was 50%, within the expected values. There were no morphological abnormalities associated to the UV-C treatment in the flies. In conclusion sterilization by UV-C rays is indicated to obtain sterile larvae destined for MDT.

Chronology of the Intrapuparial Development of the Blowfly Chrysomya albiceps (Diptera: Calliphoridae): Application in Forensic Entomology.

Insects display different patterns of development, and blow flies have one of the most specialized patterns of intrapuparial development of all. In forensic entomology, pupae can be used as a tool to estimate the minimum postmortem time interval (minPMI). We analyzed the intrapuparial development of Chrysomya albiceps (Diptera: Caloricidade), whose larvae had been fed pig lungs and reared in a climate-controlled room at 28°C day/26°C night, 70 ± 10% RH, and 12 h of photophase and monitored daily. After the third-instar larvae abandoned their diet, the process of pupariation and pupation was monitored. At pre-established times, five pupae were collected, euthanized, and fixed in 5% formaldehyde, inside polypropylene test tubes with caps. Since they were the first, they were classified as 0 h pupae. Twelve collections occurred until the emergence of the adults, at 0, 2, 4, 6, 8, 10, 24, 30, 48, 54, 72, 78, 96, and 99 h (n = 84). The fixed pupae were dissected under the microscope, with the aid of anatomical tweezers and hypodermic needles, and photographed. The stages of metamorphosis and the morphological alterations occurring during the process were identified, described, and recorded before and after pupation. These phases were: pupation, larval pupal apolysis, cryptocephalic, phanerocephalic, pharate adult, emergence, and adult. The cryptophalic phase occurred between 4 and 6 h after pupation; the phanerocephalic phase between 6 and 10 h after; the pharate adult phase between 24 and 96 h after; and the imago/emergence phase 99 h after pupation.

Calliphoridae (Diptera) Associated With Rattus rattus Carcasses in the Tijuca National Park, Rio de Janeiro, Brazil.

Forensic entomology is a complementary tool for penal procedures, mainly on estimating postmortem interval. Study of cadaveric fauna in various environments is primary as source of information to support this science. This study collected information about the fauna of Calliphoridae associated to carcasses of Rattus rattus in the Tijuca National Park, RJ. Four collections were conducted, one for each season of 2015, exposing six carcasses at georeferenced points in each collection. The carcasses were placed 550 m from the boarder and equidistant by 100 m. Five decomposition stages were identified, and 10,559 individuals of Calliphoridae belonging to 10 species were collected. The most abundant species were Hemilucilia semidiaphana (Rondani) (Diptera: Calliphoridae) and Lucilia eximia (Wiedemann) (Diptera: Calliphoridae). L. eximia was the most abundant species during the Swelling and Black Putrefaction stages, succeeded in the next stages by two species of the genera Hemilucilia. H. semidiaphana was the dominant species in the last two stages, followed by Hemilucilia segmentaria (Fabricius) (Diptera: Calliphoridae). The genus Mesembrinella (Diptera: Calliphoridae) occurred mainly during the Black putrefaction stage. Mesembrinella bellardiana (Aldrich) was more abundant, with higher occurrence during the Black putrefaction and Dry decay stages. Mesembrinella peregrina (Aldrich) occurred in the two last stages with low abundance. Huascaromusca aeneiventris (Wiedemann) (Diptera: Calliphoridae) occurred during all the observed stages, mainly during the Butyric fermentation stage. Huascaromusca purpurata (Aldrich) (Diptera: Calliphoridae) occurred only during the Dry decay stage and in low abundance. A succession pattern in the carcasses colonization was observed, providing relevant information for the resolution of criminal investigations in this environment.

Faunistic Analysis of the Families Calliphoridae and Mesembrinellidae (Diptera) at Jardim Botânico do Rio de Janeiro, Brazil.

Flies of the family Calliphoridae play a variety of ecological roles. They carry various pathogens and cause myiasis in humans and livestock, but they are useful to forensic entomology and in larval therapy. Mesembrinellidae flies, formerly classified in the family Calliphoridae, are good bioindicators of human interference in natural environments. In this study, we carried out an inventory of the Calliphoridae and Mesembrinellidae at four collecting sites within the Jardim Botânico do Rio de Janeiro. Eight traps were set, four in the arboretum (sites A and B), where the public is allowed to visit, and four at an Atlantic Forest fragment (sites C and D), open only to researchers. From July 2014 to June 2015, a total of 35,890 calliphorid flies were captured in 10 species and 145 Mesembrinellidea in 3. The greatest number of flies was found at site A and diversity was higher at site C. Chrysomya megacephala (Wiedemann) (Diptera: Calliphoridae) was the most prevalent species, being present at all sites. Mesembrinella bellardiana (Aldrich) (Diptera: Mesembrinellidae), Laneella nigripes (Guimarães) (Diptera: Mesembrinellidae), and Huascaromusca purpurata (Aldrich) (Diptera: Mesembrinellidae) were present at sites C and D, which indicates a preservation of the area because they are asynanthropic species. There were significant differences between sites A and D and sites B and D.