Effects of Terbufos (Organophosphate) on Larval Behaviour of Two Forensically Important Diptera Species: Contributions for Entomotoxicology.

Neurotoxicant compounds interfere with the behaviour and biology of insects, significantly altering their locomotion patterns. However, little is known about the effect of organophosphates, neurotoxicants for agricultural, domestic and industrial use, on the larval movement of necrophagous flies, although being responsible for frequent cases of poisoning and accidental or intentional deaths. Thus, we aimed to study the influence of Terbufos (organophosphate) on the activity and mobility patterns of Lucilia eximia (Wiedemann 1819) (Calliphoridae) and Peckia (Peckia) chrysostoma (Wiedemann 1830) (Sarcophagidae) immatures collected from rat carcasses intoxicated with 5, 10 or 20 mg/kg of Terbufos, to evaluate (i) peristaltic movements and body contractions, and (ii) distance and shape of the trajectory travelled by the larva. Behavioural parameters were analysed in loco and through videos. We observed that the presence of Terbufos altered poisoned larvae's activity and body mobility in both taxon and dose-dependent manner. Lucilia eximia larvae were more active, with greater frequency of body movements and lateral contractions when intoxicated with high and intermediate doses of Terbufos. On the other hand, P. (P.) chrysostoma immatures were less active, with fewer body and lateral contractions when intoxicated with the high dose of the compound. This work experimentally demonstrates that the presence of Terbufos can alters the mobility and movement of intoxicated necrophagous Diptera, essential components of the cadaveric fauna.

Evidence of two mitochondrial lineages and genetic variability in forensically important Lucilia eximia (Diptera: Calliphoridae) in Colombia.

Lucilia eximia (Wiedemann, 1819) (Diptera: Calliphoridae) is a blowfly with medical and forensic importance that shows genetic and color variation, however, these variations have not justified the description of new species. But in forensic entomology an accurate identification of species and subpopulations is crucial. We explored the genetic variation of L. eximia from eight localities, in five natural regions in Colombia using two mitochondrial fragments, including the standard locus for insect identification COI and the Cytb-tRNA-Ser-ND1 region. We found significant differentiation at COI and Cytb-tRNA-Ser-ND1 level, characterizing two lineages and revealing a deep and significant genetic split. High values of FST and genetic distances supported the two lineages. The origin of the divergence of L. eximia remains to discover. Examining whether the lineages have diverse ecological and biological behaviors could be a significant impact on the use of L. eximia in forensic and medical science. Our results could have relevant implications for the use of post-mortem interval estimation based on insect evidence, as well as our sequences improve the database used in DNA-based methods for identifying forensically important flies.

Myiasis in humans: Case reports in Northeastern Brazil including multispecies co-infestation by Sarcophagidae.

Myiasis is a parasitosis characterized by an infestation of living vertebrates (humans and other animals) by Diptera larvae, whose occurrence and etiological identification are still neglected by health professionals. Here we analyzed the human myiasis cases registered from 2010 to 2018 in health care units in the municipality of Natal, Northeast region of Brazil. Specifically, we aimed to: I) analyze the medical records of cases documented from 2010 to 2017; II) list the patients predisposing factors; and III) monitor the recent cases diagnosed in health units between August 2017 and March 2018 and report the taxonomic identity of the infesting species. Our data revealed that myiases mainly affect the elderly and individuals with predisposing medical conditions (e.g. senility, filariasis). Regarding the new cases, larvae of Calliphoridae (C. hominivorax (Coquerel, 1858)) and Sarcophagidae [Sarcophaga (Liopygia) ruficornis (Fabricius, 1794), Peckia (Sarcodexia) lambens (Wiedemann, 1830), and Helicobia morionella (Aldrich, 1930)] were identified. Furthermore, this was the first report of a co-infestation with three Sarcophagidae species and also the presence of H. morionella in a human wound. These results reinforce that myiasis is an underreported parasitosis, which may underestimate the ability of previously unreported fly species to feed on living human tissues in the Neotropical Region.

The Nocturnal Ovipositon Behavior of Chrysomya megacephala (Diptera: Calliphoridae) in Brazil and Its Forensic Implications.

Chrysomya megacephala (Fab. 1794) (Diptera: Calliphoridae) is a very important species for forensic entomology, mainly contributing estimations of the postmortem interval (PMI) in judicial investigations. There are some doubts about the nocturnal oviposition of these flies, which could lead to errors in the PMI calculation. This study aimed to monitor the nocturnal oviposition behavior of this species through four experimental conditions carried out in laboratory. Ten cages, each containing five males and females (n = 100), were kept in a fume hood and subjected to total darkness or to artificial light for 11 consecutive hours. Two verifications were performed to determine whether the females deposited eggs on the substrate of ~20 g of chicken gizzards per cage. The first verification occurred at 9:00 pm in nocturnal experiments and at 09:00 am in diurnal experiments. The second verification occurred at 05:00 am in nocturnal experiments and at 05:00 pm in diurnal experiments. Each experiment lasted 5 d. Chrysomya megacephala deposited eggs at night under artificial light and in total darkness, but the amount of eggs was significantly lower when compared with the daytime experiments in dark conditions and under natural light. Oviposition occurred when the average temperature was around 25°C (± 2°C) and relative humidity around 73% (± 6%). Night oviposition is a possibility which should not be ruled out. Thus, future experiments are recommended.

Lysenko and the Screwworm Fly-When Politics Interferes with Science and Public Health.

In the One Health scenario, a deep understanding of the dynamics potentially threatening the development and implementation of useful pest and vector management tools is of key importance. The New World screwworm fly, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae), is characterized by a wide host range. It acts as an important agent of myiasis in humans and warm-blooded animals in the Neotropics, and has been eliminated from a wide region through genetic methods. Of note, Serebrovsky had already proposed in 1940 the principles of autocidal control by the translocation of segments between two chromosomes, but his work was negated by Lysenko, based on the negation of Mendelian genetics. This entomological case study emphasizes the danger of politics interfering with science, a still contemporary hot issue. The negation of global warming or current pandemics are further examples of this noxious influence.

Analysis of the Effect of Cyclophosphamide and Methotrexate on Chrysomya megacephala (Diptera: Calliphoridae).

Forensic entomotoxicology investigates the effects of chemical substances in the development of scavenger insects and the reflection on estimating the minimum postmortem interval (PMI). To evaluate the impact of the chemotherapeutic drugs cyclophosphamide (CF) and methotrexate (MTX) on the postembryonic development of Chrysomya megacephala, controlled experiments were performed by rearing the larvae on spiked minced beef with different concentrations of these drugs. The results indicated that CF significantly decreased their developmental rate up to 28 h but had no significant effect on larval and adult sizes, survival rate, and sex ratio, whereas MTX decreased larval and adult sizes, survival rate, and there was a deviation in the expected sex ratio toward females in MTX-exposed larvae but had no significant impact on developmental rate. These negative interference factors should be considered in cases of suspected death of people that have undergone chemotherapy.

Insecticide activity of Baccharis dracunculifolia essential oil against Cochliomyia macellaria (Diptera: Calliphoridae).

The ethnobotanical uses of Brazilian plants for different injuries and diseases conjoined with local rich biodiversity represent an important resource for research and development. This study aimed to characterise BDEO and its in vitro activity on the third instar larvae (L3) of Cochliomyia macellaria. Groups of 20 L3 were placed on filter paper impregnated with increasing concentrations of 5-30% (v/v), equivalent to 0.79-4.77 µL/cm2, solubilised in ethanol or acetone. The major constituents of BDEO were ß-pinene (9.94%), D-limonene (9.59%), ß-nerolidol (7.93%), caryophyllene (7.69%), spathulenol (6.69), α-muurolene (6.74%) and α-pinene (5.31%). Lethal concentrations of 50% for BDEO on C. macellaria (LC50) after 24 and 48 h of exposure were 2.63 and 2.47 µL/cm2 for ethanol and 9.58 and 8.11 µL/cm2 for acetone, respectively. Furthermore, larvae cuticle abnormalities and adult deformity were observed. Our data confirm the effectiveness of BDEO as an ecofriendly product against blowflies.

Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) Oviposition Behavior in Previous Oviposition Situation.

Blowflies use their sense of smell to locate oviposition substrates and their vision to direct and assist in landing decision. Blowflies' ovipositions are usually aggregated. Chrysomya megacephala (Fabricius) females, for example, prefer to lay eggs on substrates containing fresh eggs of the same species. However, it is possible that females would be capable of evaluating the substrate, reducing the size of egg cluster in sites with a high amount of eggs or larvae, or finding another site for laying their eggs, preventing the progeny from facing high competition for space and food. The present study aimed to investigate whether females of C. megacephala could distinguish and preferentially select previously infested substrates and whether females would lay their eggs attached or separate from eggs previously laid on the substrate. Behavioral measures were conducted inside cages under laboratory conditions in free-choice tests. Data analysis confirmed that C. megacephala females are able to distinguish the presence of cospecific eggs in the substrate and preferentially select previously infested substrates for egg laying, depositing their eggs separate from pre-existing large egg masses.

Evaluation of conventional therapeutic methods versus maggot therapy in the evolution of healing of tegumental injuries in Wistar rats with and without diabetes mellitus.

Larval therapy consists on the application of sterilized carrion flies larvae, reared in laboratory, on acute, chronic, and/or infected wounds in order to promote healing. Conventional methods for treating injuries include mechanical debridement or silver-based dressings; however, they are not always effective for wound healing. Larval therapy is a feasible and safe treatment for therapeutic application and, in many cases, the only and the most recommended alternative for difficult healing injuries. Thus, this study aimed to evaluate the competence of Cochliomyia macellaria F. (Diptera: Calliphoridae) as a suitable species for therapeutic application and evaluate time and effectiveness of the types of treatments most commonly used to treat integumental injuries. C. macellaria eggs were obtained from colonies established in laboratory and sterilized prior to application. Twenty-five larvae were applied for each centimeter squared of lesion. Lesions were induced in 24 Wistar rats; type 1 diabetes mellitus was induced in 12 of them. Animals were divided in four groups with three individuals each, being denominated: larval therapy, larval therapy associated with foam dressing with silver release, mechanical debridement with foam dressing silver and control group, without treatment. All treatments were applied once and held for 24 h. Medical application of larvae was found to be safe, as only dead tissue was removed, and efficient to accelerate healing process when compared to other treatments.

Induced Cannibalism in Experimental Populations of the Forensic Indicator Chrysomya putoria Wiedemann (Diptera: Calliphoridae).

We analyzed the behavior of third-instars of Chrysomya putoria as potential cannibals in experimental populations. Cannibalism rates were evaluated in three settings observed for 3, 6, 9, and 24 h, placing injured and uninjured larvae of C. putoria together. Our data heavily support that C. putoria larvae behave as cannibals when induced by a wound in another larva, and also after starving for 24 h. The probability of cannibalism increased as a function of time, both in no-choice and in choice experiments evidencing that time is a determining factor for cannibalism induction in C. putoria. However, the treatment combining injured with uninjured larvae showed the highest probability of cannibalism. These results suggest that C. putoria larvae may cannibalize under scarcity of food over long time or the presence of injured larvae. This study is useful to understand the behavior of C. putoria feeding on ephemeral substrates such as carrion or corpses and brings relevant and significant contribution to population ecology of blowflies and also forensic entomology.

Effect of bait decomposition on the attractiveness to species of Diptera of veterinary and forensic importance in a rainforest fragment in Brazil.

Insects associated with carrion can have parasitological importance as vectors of several pathogens and causal agents of myiasis to men and to domestic and wild animals. We tested the attractiveness of animal baits (chicken liver) at different stages of decomposition to necrophagous species of Diptera (Calliphoridae, Fanniidae, Muscidae, Phoridae and Sarcophagidae) in a rainforest fragment in Brazil. Five types of bait were used: fresh and decomposed at room temperature (26 °C) for 24, 48, 72 and 96 h. A positive correlation was detected between the time of decomposition and the abundance of Calliphoridae and Muscidae, whilst the abundance of adults of Phoridae decreased with the time of decomposition. Ten species of calliphorids were registered, of which Chrysomya albiceps, Chrysomya megacephala and Chloroprocta idioidea showed a positive significant correlation between abundance and decomposition. Specimens of Sarcophagidae and Fanniidae did not discriminate between fresh and highly decomposed baits. A strong female bias was registered for all species of Calliphoridae irrespective of the type of bait. The results reinforce the feasibility of using animal tissues as attractants to a wide diversity of dipterans of medical, parasitological and forensic importance in short-term surveys, especially using baits at intermediate stages of decomposition.

Mesembrinellinae (Diptera: Calliphoridae) to edge effects in the Tinguá Biological Reserve, Rio de Janeiro, Brazil / Mesembrinellinae (Diptera: Calliphoridae) na Reserva Biológica do Tinguá, Rio de Janeiro, Brasil

In this study we describe the diversity of Mesembrinelinae in a biological reserve in the city of Nova Iguaçu, State of Rio de Janeiro. Traps containing sardines were distributed seasonally, at four sites: Site A (22° 58.788 S, 43° 43.459 W), in a forest edge, and sites B (22° 58.523 S, 43° 44.540 W), C (22° 58.350 S, 43° 44.678 W), and D (22° 34.865 S, 43° 27.063 W), located 1,000m, 500m, and 2,000m respectively, inwards from the edge. A total of 2,150 individuals of Mesembrinellinae were collected, representing ten species. Laneela nigripes Guimarães, 1977 was the most abundant species, followed by Mesembrinella bellardiana Aldrich, 1922, Eumesembrinella cyaneicincta (Surcouf, 1919) and Mesembrinella semihyalina Mello, 1967. These species were common and constant during the study period. Mesembrinella batesi Aldrich, 1922, Eumesembrinella quadrilineata (Fabricius, 1805) and Huascaromusca aeneiventris (Wiedmann, 1830) were the less abundant flies, being considered rare and accidental. Eumesembrinella besnoiti (Seguy, 1925) was rare and accessory. Eumesembrinella cyaneicincta, M. bellardiana, M. semihyalina and M. bicolor were mostly collected in site B, while L. nigripes was mostly collected in site C. The edge effect was not evident since the four sites showed similar populations. Site B showed a strong positive relationship between abundance and richness, in site C the correlation was positive and weak, and there was no correlation in A and D. The highest abundance of specimens was recorded during autumn and winter. These flies occurred from the edge up to 2,000m inside the forest.(AU)

Nesse estudo, descrevemos a diversidade de Mesembrinelinae, em uma reserva biológica na cidade de Nova Iguaçu, Estado do Rio de Janeiro. Armadilhas contendo sardinha foram distribuídas sazonalmente em quatro pontos: Ponto A (22° 58.788 S, 43° 43.459 W), na borda da mata B (22° 58.523 S, 43° 44.540 W), C (22° 58.350 S, 43° 44.678 W), e D (22° 34.865 S, 43º 27.063 W), localizados a 1.000m, 500m e 2.000m para o interior da borda respectivamente. Foram coletados 2.159 exemplares de Mesembrinellinae, com um total de dez espécies. Laneela nigripes Guimarães, 1977 foi a espécie mais abundante, seguida por Mesembrinella bellardiana Aldrich, 1922, Eumesembrinella cyaneicincta (Surcouf, 1919) e Mesembrinella semihyalina Mello, 1967. Essas espécies foram consideradas comuns e contantes durante o período de estudo. Mesembrinella batesi Aldrich, 1922, Eumesembrinella quadrilineata Fabricius, 1805 e Huascaromusca aeneiventris (Wiedmann, 1830) foram as moscas com as menores abundâncias, sendo consideradas raras e acidentais, Eumesembrinella besnoiti (Seguy, 1925) foi rara e acessória. Eumesembrinella cyaneicincta, M. bellardiana, M. semihyalina e M. bicolor foram as mais coletadas no ponto B, enquanto L. nigripes foi a mais coletada no ponto C. Os efeitos de borda não foram evidentes porque os quatro pontos apresentaram populações similares. O ponto B apresentou uma relação forte e positiva entre a abundância e a riqueza, no ponto C a correlação foi positiva e fraca e não houve correlação nos pontos A e D. A maior abundância de indivíduos foi registrada nas estações do outono e inverno. Espécimes da subfamília Mesembrinellinae ocorreram da borda até 2.000 metros para o interior da mata.(AU)

Mesembrinellinae (Diptera: Calliphoridae) to edge effects in the Tinguá Biological Reserve, Rio de Janeiro, Brazil

Abstract In this study we describe the diversity of Mesembrinelinae in a biological reserve in the city of Nova Iguaçu, State of Rio de Janeiro. Traps containing sardines were distributed seasonally, at four sites: Site A (22° 58.788 S, 43° 43.459 W), in a forest edge, and sites B (22° 58.523 S, 43° 44.540 W), C (22° 58.350 S, 43° 44.678 W), and D (22° 34.865 S, 43° 27.063 W), located 1,000m, 500m, and 2,000m respectively, inwards from the edge. A total of 2,150 individuals of Mesembrinellinae were collected, representing ten species. Laneela nigripes Guimarães, 1977 was the most abundant species, followed by Mesembrinella bellardiana Aldrich, 1922, Eumesembrinella cyaneicincta (Surcouf, 1919) and Mesembrinella semihyalina Mello, 1967. These species were common and constant during the study period. Mesembrinella batesi Aldrich, 1922, Eumesembrinella quadrilineata (Fabricius, 1805) and Huascaromusca aeneiventris (Wiedmann, 1830) were the less abundant flies, being considered rare and accidental. Eumesembrinella besnoiti (Seguy, 1925) was rare and accessory. Eumesembrinella cyaneicincta, M. bellardiana, M. semihyalina and M. bicolor were mostly collected in site B, while L. nigripes was mostly collected in site C. The edge effect was not evident since the four sites showed similar populations. Site B showed a strong positive relationship between abundance and richness, in site C the correlation was positive and weak, and there was no correlation in A and D. The highest abundance of specimens was recorded during autumn and winter. These flies occurred from the edge up to 2,000m inside the forest.

Resumo Nesse estudo, descrevemos a diversidade de Mesembrinelinae, em uma reserva biológica na cidade de Nova Iguaçu, Estado do Rio de Janeiro. Armadilhas contendo sardinha foram distribuídas sazonalmente em quatro pontos: Ponto A (22° 58.788 S, 43° 43.459 W), na borda da mata B (22° 58.523 S, 43° 44.540 W), C (22° 58.350 S, 43° 44.678 W), e D (22° 34.865 S, 43º 27.063 W), localizados a 1.000m, 500m e 2.000m para o interior da borda respectivamente. Foram coletados 2.159 exemplares de Mesembrinellinae, com um total de dez espécies. Laneela nigripes Guimarães, 1977 foi a espécie mais abundante, seguida por Mesembrinella bellardiana Aldrich, 1922, Eumesembrinella cyaneicincta (Surcouf, 1919) e Mesembrinella semihyalina Mello, 1967. Essas espécies foram consideradas comuns e contantes durante o período de estudo. Mesembrinella batesi Aldrich, 1922, Eumesembrinella quadrilineata Fabricius, 1805 e Huascaromusca aeneiventris (Wiedmann, 1830) foram as moscas com as menores abundâncias, sendo consideradas raras e acidentais, Eumesembrinella besnoiti (Seguy, 1925) foi rara e acessória. Eumesembrinella cyaneicincta, M. bellardiana, M. semihyalina e M. bicolor foram as mais coletadas no ponto B, enquanto L. nigripes foi a mais coletada no ponto C. Os efeitos de borda não foram evidentes porque os quatro pontos apresentaram populações similares. O ponto B apresentou uma relação forte e positiva entre a abundância e a riqueza, no ponto C a correlação foi positiva e fraca e não houve correlação nos pontos A e D. A maior abundância de indivíduos foi registrada nas estações do outono e inverno. Espécimes da subfamília Mesembrinellinae ocorreram da borda até 2.000 metros para o interior da mata.

Mesembrinellinae (Diptera: Calliphoridae) to edge effects in the Tinguá Biological Reserve, Rio de Janeiro, Brazil / Mesembrinellinae (Diptera: Calliphoridae) na Reserva Biológica do Tinguá, Rio de Janeiro, Brasil

Abstract In this study we describe the diversity of Mesembrinelinae in a biological reserve in the city of Nova Iguaçu, State of Rio de Janeiro. Traps containing sardines were distributed seasonally, at four sites: Site A (22° 58.788’ S, 43° 43.459’ W), in a forest edge, and sites B (22° 58.523’ S, 43° 44.540’ W), C (22° 58.350’ S, 43° 44.678’ W), and D (22° 34.865’ S, 43° 27.063’ W), located 1,000m, 500m, and 2,000m respectively, inwards from the edge. A total of 2,150 individuals of Mesembrinellinae were collected, representing ten species. Laneela nigripes Guimarães, 1977 was the most abundant species, followed by Mesembrinella bellardiana Aldrich, 1922, Eumesembrinella cyaneicincta (Surcouf, 1919) and Mesembrinella semihyalina Mello, 1967. These species were common and constant during the study period. Mesembrinella batesi Aldrich, 1922, Eumesembrinella quadrilineata (Fabricius, 1805) and Huascaromusca aeneiventris (Wiedmann, 1830) were the less abundant flies, being considered rare and accidental. Eumesembrinella besnoiti (Seguy, 1925) was rare and accessory. Eumesembrinella cyaneicincta, M. bellardiana, M. semihyalina and M. bicolor were mostly collected in site B, while L. nigripes was mostly collected in site C. The edge effect was not evident since the four sites showed similar populations. Site B showed a strong positive relationship between abundance and richness, in site C the correlation was positive and weak, and there was no correlation in A and D. The highest abundance of specimens was recorded during autumn and winter. These flies occurred from the edge up to 2,000m inside the forest.

Resumo Nesse estudo, descrevemos a diversidade de Mesembrinelinae, em uma reserva biológica na cidade de Nova Iguaçu, Estado do Rio de Janeiro. Armadilhas contendo sardinha foram distribuídas sazonalmente em quatro pontos: Ponto A (22° 58.788’ S, 43° 43.459’ W), na borda da mata B (22° 58.523’ S, 43° 44.540’ W), C (22° 58.350’ S, 43° 44.678’ W), e D (22° 34.865’ S, 43º 27.063’ W), localizados a 1.000m, 500m e 2.000m para o interior da borda respectivamente. Foram coletados 2.159 exemplares de Mesembrinellinae, com um total de dez espécies. Laneela nigripes Guimarães, 1977 foi a espécie mais abundante, seguida por Mesembrinella bellardiana Aldrich, 1922, Eumesembrinella cyaneicincta (Surcouf, 1919) e Mesembrinella semihyalina Mello, 1967. Essas espécies foram consideradas comuns e contantes durante o período de estudo. Mesembrinella batesi Aldrich, 1922, Eumesembrinella quadrilineata Fabricius, 1805 e Huascaromusca aeneiventris (Wiedmann, 1830) foram as moscas com as menores abundâncias, sendo consideradas raras e acidentais, Eumesembrinella besnoiti (Seguy, 1925) foi rara e acessória. Eumesembrinella cyaneicincta, M. bellardiana, M. semihyalina e M. bicolor foram as mais coletadas no ponto B, enquanto L. nigripes foi a mais coletada no ponto C. Os efeitos de borda não foram evidentes porque os quatro pontos apresentaram populações similares. O ponto B apresentou uma relação forte e positiva entre a abundância e a riqueza, no ponto C a correlação foi positiva e fraca e não houve correlação nos pontos A e D. A maior abundância de indivíduos foi registrada nas estações do outono e inverno. Espécimes da subfamília Mesembrinellinae ocorreram da borda até 2.000 metros para o interior da mata.

Detection and preliminary physico-chemical properties of antimicrobial components in the native excretions/secretions of three species of Chrysomya (Diptera, Calliphoridae) in Brazil.

Antibiotic-resistant bacteria in hospitals and communities increasingly threaten public health in Brazil and the rest of the World. There is an urgent need for additional antimicrobial drugs. Calliphorid blowfly larvae are a rich source of antimicrobial factors but the potential of Neotropical species has been neglected. This preliminary study evaluates the antimicrobial activity of the native excretions/secretions of larvae of three species of Brazilian calliphorids, Chrysomya megacephala, Chrysomya albiceps and Chrysomya putoria. Native excretions/secretions were collected from third instar larvae, sterile filtered and tested for antibacterial activity against Staphylococcus aureus 9518, Escherichia coli K12 4401 and Serratia marcescens 365. Turbidometric assays were made in micro-plates, using an ELISA reader, with readings taken up to 22 h. Bacterial suspensions at the start and end of each experiment were also serially diluted, spread on nutrient agar plates and then colony forming units counted. The physico-chemical characteristics of the native excretions/secretions were also tested by freezing/thawing, boiling, and protease digestion. The native excretions/secretions of larvae from these three Chrysomya species significantly inhibited bacterial growth. Therefore, Brazilian calliphorid flies could potentially provide new classes of antibiotics.

Phenotypic polymorphism of Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae) may lead to species misidentification.

Species identification is an essential step in the progress and completion of work in several areas of biological knowledge, but it is not a simple process. Due to the close phylogenetic relationship of certain species, morphological characters are not always sufficiently distinguishable. As a result, it is necessary to combine several methods of analysis that contribute to a distinct categorization of taxa. This study aimed to raise diagnostic characters, both morphological and molecular, for the correct identification of species of the genus Chrysomya (Diptera: Calliphoridae) recorded in the New World, which has continuously generated discussion about its taxonomic position over the last century. A clear example of this situation was the first record of Chrysomya rufifacies in Brazilian territory in 2012. However, the morphological polymorphism and genetic variability of Chrysomya albiceps studied here show that both species (C. rufifacies and C. albiceps) share very similar character states, leading to misidentification and subsequent registration error of species present in our territory. This conclusion is demonstrated by the authors, based on a review of the material deposited in major scientific collections in Brazil and subsequent molecular and phylogenetic analysis of these samples. Additionally, we have proposed a new taxonomic key to separate the species of Chrysomya found on the American continent, taking into account a larger number of characters beyond those available in current literature.

Evaluation of the influence of the antibiotic ciprofloxacin in the development of an Old World screwworm fly, Chrysomya putoria.

Chrysomya putoria (Wiedemann) (Diptera: Calliphoridae), an Old World screwworm fly, is a species with potential for maggot therapy practice and has been described in myiasis and forensic entomology studies. The objective of the present study was to assess the action of different ciprofloxacin concentrations on the growth and development of C. putoria. First instar maggots of the third generation were raised on 60 g of chicken gizzard homogenate in 65% agar diet and received ciprofloxacin chloridrate. Each concentration of the antibiotic tested (3.33 µg/mL, 6.66 µg/mL, and 13.33 µg/mL) and the control (no antibiotic) were replicated four times (40 maggots/replication). The control received distilled water instead of the antibiotic. Maggots were kept in an acclimatized chamber at 30° C during the day and 28° C at night, with 70 + 10% RH and a 14:10 L:D photoperiod. They were weighed in batches of five and stored in test tubes sealed with nylon fabric and elastic. Microsoft Excel and STAT were used for the analysis. The variation among the maggot weight means and the duration of the maggot stage, pupal stage, and time to total development (neolarvae to adult) were analyzed by Student's t-test (α= 5%). The viabilities and the normality rates were compared using ANOVA, and the expected sex ratio frequency was tested by the chisquared test (χ(2)). There was no significant difference among the four treatments regarding mean individual maggot weight, mean duration of the maggot inoculation until abandonment, the duration of the maggot and pupal stages, and the total duration of all stages. The sex ratios found in the four treatments did not differ from the expected. Only treatment 2 (6.66 µg/mL concentration of ciprofloxacin) differed significantly from the control in maggot and total viability. The antibiotic did not seem to alter C. putoria development in the postembryonic period.

Comparative morphology of the spermathecae of some species of Chrysomya Robineau-Desvoidy and Cochliomyia Townsend (Diptera, Calliphoridae)

Comparative morphology of the spermathecae of some species of Chrysomya Robineau-Desvoidy and Cochliomyia Townsend (Diptera, Calliphoridae). Little is known about the morphology of the chitinized structures of the spermathecae of the Calliphoridae. In this work, the spermathecae of Chrysomya albiceps Wiedemann, 1819, C. megacephala Fabricius, 1794, Cochliomyia macellaria Fabricius, 1775 and C. hominivorax Coquerel, 1858 are described and illustrated. The occurrence in one species of four spermathecae, an atypical form for blow flies, was recorded for the first time. The analysis of these structures will allow a better understanding of this group as well as provide taxonomic characters for future phylogenetic studies.

Morfologia comparada das espermatecas de espécies de Chrysomya Robineau-Desvoidy e Cochliomyia Townsend (Diptera, Calliphoridae). Pouco se conhece sobre a morfologia das estruturas quitinizadas das espermatecas de Calliphoridae. Nesse trabalho as espermatecas de Chrysomya albiceps Wiedemann, 1819, C. megacephala Fabricius, 1794, Cochliomyia macellaria Fabricius, 1775 e C. hominivorax Coquerel, 1858 são descritas e ilustradas. Foi registrada pela primeira vez a ocorrência em uma espécie com quatro espermatecas, uma forma atípica em califorídeos. A análise dessas estruturas possibilitará uma melhor compreensão do grupo e fornecerá caracteres taxonômicos para futuros estudos filogenéticos.

Necrophagous species of Diptera and Coleoptera in northeastern Brazil: state of the art and challenges for the Forensic Entomologist / Espécies necrófagas de Diptera e Coleoptera na Região Nordeste do Brasil: estado da arte e desafios para o Entomologista Forense

Inventories on necrophagous insects carried out in Brazil encompass mostly species from the southeastern and central-western regions of the country. This review aims to produce the first checklist of necrophagous Diptera and Coleoptera species of forensic relevance in northeastern Brazil, an area that concentrates high rates of homicides. We performed a literature survey on scientific articles, theses and dissertations regarding necrophagous insect species in the region, and contacted scientists who develop research on forensic entomology. Fifty-two species of Diptera belonging to eight families with previous record of necrophagy were reported in the region: Sarcophagidae, Calliphoridae, Muscidae, Fanniidae, Piophilidae, Phoridae, Anthomyiidae and Stratiomyidae. Coleopteran species from six families of forensic relevance were registered, although taxonomical identification remained superficial. Bait traps were the most frequent methodology used, followed by collection on animal carcasses. Seven Dipteran species from two families were registered on human cadavers. All species had been previously reported in other Brazilian states and/or other countries, although none has been effectively used in legal procedures in the region. The status of research on forensic entomology in northeastern Brazil is incipient, and the checklist produced here contributes to the knowledge on the local diversity of necrophagous insects.

Inventários de insetos necrófagos no Brasil abordam em sua maioria espécies das regiões Sudeste e Centro-Oeste do país. Esta revisão visa apresentar um checklist de espécies necrófagas de Diptera e Coleoptera de importância forense no Nordeste brasileiro, uma área que concentra elevadas taxas de homicídio. Nós conduzimos uma revisão bibliográfica consultando artigos, teses e dissertações sobre espécies necrófagas na região, e contatamos cientistas que desenvolvem pesquisa em entomologia forense. Cinquenta e duas espécies de Diptera pertencentes a oito famílias com registro prévio de necrofagia foram registrados na região: Sarcophagidae, Calliphoridae, Muscidae, Fanniidae, Piophilidae, Phoridae, Anthomyiidae e Stratiomyidae. Espécies de Coleoptera de seis famílias de importância forense foram listadas, embora a identificação taxonômica tenha sido superficial. Armadilhas contendo iscas foram o método de coleta mais frequente, seguidas de coleta sobre carcaças animais. Sete espécies de Diptera pertencentes a duas famílias foram registradas sobre cadáveres humanos. Todas as espécies haviam sido previamente registradas em outros estados brasileiros e/ou em outros países, embora nenhuma tenha sido efetivamente utilizada em procedimentos legais na região. O status da pesquisa em entomologia forense no Nordeste do Brasil é incipiente, e o checklist apresentado neste trabalho contribui para sistematizar o conhecimento sobre a diversidade local de insetos necrófagos.

Diversity of Calliphoridae and Sarcophagidae (Diptera, Oestroidea) in continuous forest and gaps at different stages of regeneration in the Urucu oilfield in western Brazilian Amazonia

Diversity of Calliphoridae and Sarcophagidae (Diptera, Oestroidea) in continuous forest and gaps at different stages of regeneration in the Urucu oilfield in western Brazilian Amazonia. The diversity of Calliphoridae and Sarcophagidae in continuous forest and gaps at different stages of regeneration was studied in the Urucu river basin, in Coari, state of Amazonas, Brazil. The flies were collected at 16 sampling points, 12 in gaps at different stages of regeneration (early _ C1, mid- C2 and late successional _ C3) and four in continuous forest _ MT. The diversity of blowflies was similar in the two less regenerated habitats (C1 and C2), and lower than that in the late successional (C3) and continuous forests (MT). By contrast, the diversity of flesh flies was much higher in all three types of gaps (C1, C2 and C3) in comparison with continuous forest (MT). Ordination (NMDS) and similarity (ANOSIM) analyses revealed that the blowflies communities were grouped by habitat type, which affected species composition more than diversity. Analysis of the flesh flies revealed two main groupings, gaps (C1, C2 and C3) and continuous forest (MT), with no evidence of any influence of successional stage on the diversity of the community.

Diversidade de Calliphoridae e Sarcophagidae (Diptera, Oestroidea) em florestas contínuas e clareiras com diferentes estágios de regeneração no campo petrolífero Urucu no oeste da Amazônia brasileira. A diversidade de Calliphoridae e Sarcophagidae em florestas contínuas e clareiras com diferentes estágios de regeneração foi avaliada na bacia do Rio Urucu, Coari, Estado do Amazonas, Brasil. As moscas foram coletadas em 16 áreas amostrais, 12 de clareira, em diferentes estágios de regeneração (pouco recuperadas _ C1, semi-recuperadas _ C2, bem recuperadas _ C3) e quatro em floresta contínua _ MT). A diversidade de Calliphoridae foi similar nos dois habitats menos recuperados (C1 e C2), sendo menores do que as dos habitats de clareiras bem recuperadas (C3) e florestas contínuas (MT). Entretanto, a diversidade de Sarcophagidae foi muito maior nos três tipos de clareiras (C1, C2 e C3) quando comparada com a floresta contínua (MT). As análises de ordenação (NMDS) e de similaridade (ANOSIM) demonstraram que as comunidades de Calliphoridae foram agrupadas conforme os quatro tipos de ambientes, e que a composição de espécies foi mais afetada do que a diversidade. A análise de Sarcophagidae revelou a formação de dois grupos, clareiras (C1, C2 e C3) e florestas contínuas (MT), sem evidência de qualquer influencia dos estágios de recuperação das clareiras sobre a diversidade da comunidade.