Reflectance-based determination of age and species of blowfly puparia.

Science.gov (United States)

Voss, Sasha C; Magni, Paola; Dadour, Ian; Nansen, Christian

2017-01-01

Forensic entomology is primarily concerned with the estimation of time since death and involves determination of the age of immature insects colonising decomposing remains. Accurate age determination of puparia is usually accomplished by dissection, which means destructive sampling of evidence. As part of improving abilities to correctly identify species and developmental age, it is highly desirable to have available non-destructive methods. In this study, we acquired external hyperspectral imaging (HSI) data (77 spectral bands, 389-892 nm) from the dorsal and ventral sides of individual puparia of two species of blowfly (Diptera: Calliphoridae), Calliphora dubia Macquart 1855 and Chrysomya rufifacies Macquart 1842. Puparia were dissected to determine the presence/absence of eight internal morphological development characteristics (legs, wings, labella, abdominal segments, antennae, thoracic bristles, orbital/facial bristles and eye colour and arista). Based on linear discriminant analysis and independent validation of HSI data, reflectance features from puparia could be used to successfully (1) distinguish the two species (classification accuracy = 92.5 %), (2) differentiate dorsal and ventral sides of puparia (classification accuracy C. dubia = 81.5 %; Ch. rufifacies = 89.2 %) and (3) predict the presence of these morphological characteristics and therefore the developmental stage of puparia (average classification accuracy using dorsal imaging: C. dubia = 90.3 %; Ch. rufifacies = 94.0 %). The analytical approach presented here provides proof of concept for a direct puparial age relationship (i.e. days since the onset of pupation) between external puparial reflectance features and internal morphological development. Furthermore, this approach establishes the potential for further refinement by using a non-invasive technique to determine the age and developmental stage of blowflies of forensic importance.

Dynamic feedback circuits function as a switch for shaping a maturation-inducing steroid pulse in Drosophila

DEFF Research Database (Denmark)

Møller, Morten Erik; Danielsen, Erik Thomas; Herder, Rachel

2013-01-01

. Remarkably, our study shows that the same well-defined genetic program that stimulates a systemic downstream response to ecdysone is also utilized upstream to set the duration and amplitude of the ecdysone pulse. Activation of this switch-like mechanism ensures a rapid, self-limiting PG response......Steroid hormones trigger the onset of sexual maturation in animals by initiating genetic response programs that are determined by steroid pulse frequency, amplitude and duration. Although steroid pulses coordinate growth and timing of maturation during development, the mechanisms generating...... of hormone synthesis, the two key parameters determining pulse shape (amplitude and duration). We show that ecdysone has a positive-feedback effect on the PG, rapidly amplifying its own synthesis to trigger pupariation as the onset of maturation. During the prepupal stage, a negative-feedback signal ensures...

DNA extraction and barcode identification of development stages of forensically important flies in the Czech Republic.

Science.gov (United States)

Olekšáková, Tereza; Žurovcová, Martina; Klimešová, Vanda; Barták, Miroslav; Šuláková, Hana

2018-04-01

Several methods of DNA extraction, coupled with 'DNA barcoding' species identification, were compared using specimens from early developmental stages of forensically important flies from the Calliphoridae and Sarcophagidae families. DNA was extracted at three immature stages - eggs, the first instar larvae, and empty pupal cases (puparia) - using four different extraction methods, namely, one simple 'homemade' extraction buffer protocol and three commercial kits. The extraction conditions, including the amount of proteinase K and incubation times, were optimized. The simple extraction buffer method was successful for half of the eggs and for the first instar larval samples. The DNA Lego Kit and DEP-25 DNA Extraction Kit were useful for DNA extractions from the first instar larvae samples, and the DNA Lego Kit was also successful regarding the extraction from eggs. The QIAamp DNA mini kit was the most effective; the extraction was successful with regard to all sample types - eggs, larvae, and pupari.

Effects of larval population density on rates of development and interactions between two species of Chrysomya (Diptera: Calliphoridae) in laboratory culture.

Science.gov (United States)

Goodbrod, J R; Goff, M L

1990-05-01

Rearing of Chrysomya megacephala (F.) and Chrysomya rufifacies (Macquart) in pure cultures at seven different population densities (larvae per gram of liver) demonstrated an inverse relationship between density and the duration of the larval stage. In pure cultures, larval mortality rates decreased with increasing density until an optimum density was reached (8 larvae/g liver for C. megacephala and 10 larvae/g liver for C. rufifacies), then decreased directly with density. Puparial and adult weights varied inversely with density for both species in pure cultures. Internal feeding mass temperatures were above ambient temperatures for all cultures, with maximum temperatures recorded in cultures with 20 and 40 larvae/g liver for G. rufifacies and C. megacephala, respectively. In paired encounters, larvae of C. rufifacies were cannibalistic and predatory on C. megacephala larvae after the first instar. In mixed cultures of these two species, the larval mortality of C. rufifacies remained relatively stable, whereas the larval mortality of C. megacephala increased directly with population density.

Developmental Times of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) at Constant Temperatures and Applications in Forensic Entomology.

Science.gov (United States)

Yang, Yong-Qiang; Li, Xue-Bo; Shao, Ru-Yue; Lyu, Zhou; Li, Hong-Wei; Li, Gen-Ping; Xu, Lyu-Zi; Wan, Li-Hua

2016-09-01

The characteristic life stages of infesting blowflies (Calliphoridae) such as Chrysomya megacephala (Fabricius) are powerful evidence for estimating the death time of a corpse, but an established reference of developmental times for local blowfly species is required. We determined the developmental rates of C. megacephala from southwest China at seven constant temperatures (16-34°C). Isomegalen and isomorphen diagrams were constructed based on the larval length and time for each developmental event (first ecdysis, second ecdysis, wandering, pupariation, and eclosion), at each temperature. A thermal summation model was constructed by estimating the developmental threshold temperature D0 and the thermal summation constant K. The thermal summation model indicated that, for complete development from egg hatching to eclosion, D0 = 9.07 ± 0.54°C and K = 3991.07 ± 187.26 h °C. This reference can increase the accuracy of estimations of postmortem intervals in China by predicting the growth of C. megacephala. © 2016 American Academy of Forensic Sciences.

Gene Expression Associated with Early and Late Chronotypes in Drosophila melanogaster

Directory of Open Access Journals (Sweden)

Mirko ePegoraro

2015-05-01

Full Text Available The circadian clock provides the temporal framework for rhythmic behavioural and metabolic functions. In the modern era of industrialization, work and social pressures, the clock function is often jeopardized, resulting in adverse and chronic effects on health. Understanding circadian clock function, particularly individual variation in diurnal phase preference (chronotype, and the molecular mechanisms underlying such chronotypes may lead to interventions that could abrogate clock dysfunction and improve human (and animal health and welfare. Our preliminary studies suggested that fruitflies, like humans, can be classified as early rising ‘larks’ or late rising ‘owls’, providing a convenient model system for these types of studies. We have identified strains of flies showing increased preference for morning emergence (Early or E from the pupal case, or more pronounced preference for evening emergence (Late or L. We have sampled pupae the day before eclosion (4th day after pupariation at 4 h intervals in the E and L strains, and examined differences in gene expression by RNAseq. We have identified differentially expressed transcripts between the E and L strains which provide candidate genes for studies of Drosophila chronotypes and their human orthologues.

Nanos-mediated repression of hid protects larval sensory neurons after a global switch in sensitivity to apoptotic signals.

Science.gov (United States)

Bhogal, Balpreet; Plaza-Jennings, Amara; Gavis, Elizabeth R

2016-06-15

Dendritic arbor morphology is a key determinant of neuronal function. Once established, dendrite branching patterns must be maintained as the animal develops to ensure receptive field coverage. The translational repressors Nanos (Nos) and Pumilio (Pum) are required to maintain dendrite growth and branching of Drosophila larval class IV dendritic arborization (da) neurons, but their specific regulatory role remains unknown. We show that Nos-Pum-mediated repression of the pro-apoptotic gene head involution defective (hid) is required to maintain a balance of dendritic growth and retraction in class IV da neurons and that upregulation of hid results in decreased branching because of an increase in caspase activity. The temporal requirement for nos correlates with an ecdysone-triggered switch in sensitivity to apoptotic stimuli that occurs during the mid-L3 transition. We find that hid is required during pupariation for caspase-dependent pruning of class IV da neurons and that Nos and Pum delay pruning. Together, these results suggest that Nos and Pum provide a crucial neuroprotective regulatory layer to ensure that neurons behave appropriately in response to developmental cues. © 2016. Published by The Company of Biologists Ltd.

A High Throughput Ambient Mass Spectrometric Approach to Species Identification and Classification from Chemical Fingerprint Signatures

Science.gov (United States)

Musah, Rabi A.; Espinoza, Edgard O.; Cody, Robert B.; Lesiak, Ashton D.; Christensen, Earl D.; Moore, Hannah E.; Maleknia, Simin; Drijfhout, Falko P.

2015-01-01

A high throughput method for species identification and classification through chemometric processing of direct analysis in real time (DART) mass spectrometry-derived fingerprint signatures has been developed. The method entails introduction of samples to the open air space between the DART ion source and the mass spectrometer inlet, with the entire observed mass spectral fingerprint subjected to unsupervised hierarchical clustering processing. A range of both polar and non-polar chemotypes are instantaneously detected. The result is identification and species level classification based on the entire DART-MS spectrum. Here, we illustrate how the method can be used to: (1) distinguish between endangered woods regulated by the Convention for the International Trade of Endangered Flora and Fauna (CITES) treaty; (2) assess the origin and by extension the properties of biodiesel feedstocks; (3) determine insect species from analysis of puparial casings; (4) distinguish between psychoactive plants products; and (5) differentiate between Eucalyptus species. An advantage of the hierarchical clustering approach to processing of the DART-MS derived fingerprint is that it shows both similarities and differences between species based on their chemotypes. Furthermore, full knowledge of the identities of the constituents contained within the small molecule profile of analyzed samples is not required. PMID:26156000

Ionization with accelerated high energy electrons as quarantine treatment against Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) in citrus fruits

International Nuclear Information System (INIS)

Alonso, M.; Río, M.A. del; Jacas, J.

2002-01-01

Ceratitis capitata is a quarantine pest. Spanish citrus exports to countries such as the USA or Japan are subjected to a mandatory quarantine treatment consisting of exposure of fruits to a low temperatures. Some citrus (''Fino'' lemon, ''Fortune'' mandarin) are very sensitive to this kind of treatment and can not be treated this way. Therefore, alternative treatments are necessary. In this study, high energy electrons were investigated as an alternative quarantine treatment against C. capitata in citrus. Survival of the different instars (egg to old pupae) of C. capitata reared in an artificial medium was assessed when exposed to different doses between 0 and 1 kGy. Both pupariation and adult emergence were almost prevented at 0.25 kGy, and no viable adults were obtained at 0.50 kGy. When artificially infested fruits (in both ''Fino'' lemon and ''Fortune'' mandarin) were exposed to 1 kGy, 100% mortality was obtained. Finally, quality (texture, color index, maturity index, juice yield, ethanol and acetaldehyde contents, physiological alterations and organoleptic characteristics) of irradiated (1 kGy) and non irradiated fruit were compared. High energy electron irradiation resulted in unacceptable damage to ''Fortune'' mandarin, but quality of ''Fino'' lemon resulted unaltered even when evaluated one month after irradiation. Therefore high energy electrons could be a useful alternative to cold quarantine treatment for ''Fino'' lemons. (author) [es

DNA degradation and genetic analysis of empty puparia: genetic identification limits in forensic entomology.

Science.gov (United States)

Mazzanti, Morena; Alessandrini, Federica; Tagliabracci, Adriano; Wells, Jeffrey D; Campobasso, Carlo P

2010-02-25

Puparial cases are common remnants of necrophagous flies in crime investigations. They usually represent the longest developmental time and, therefore, they can be very useful for the estimation of the post-mortem interval (PMI). However, before any PMI estimate, it is crucial to identify the species of fly eclosed from each puparium associated with the corpse. Morphological characteristics of the puparium are often distinctive enough to permit a species identification. But, even an accurate morphological analysis of empty puparia cannot discriminate among different species of closely related flies. Furthermore, morphological identification may be impossible if the fly puparia are poorly preserved or in fragments. This study explores the applicability of biomolecular techniques on empty puparia and their fragments for identification purposes. A total of 63 empty puparia of necrophagous Diptera resulting from forensic casework were examined. Samples were divided into three groups according to size, type and time of eclosion in order to verify whether the physical characteristics and puparia weathering can influence the amount of DNA extraction. The results suggest that a reliable genetic identification of forensically important flies may also be performed from empty puparia and/or their fragments. However, DNA degradation can deeply compromise the genetic analysis since the older the fly puparia, the smaller are the amplified fragments. 2009 Elsevier Ireland Ltd. All rights reserved.

Virtual forensic entomology: improving estimates of minimum post-mortem interval with 3D micro-computed tomography.

Science.gov (United States)

Richards, Cameron S; Simonsen, Thomas J; Abel, Richard L; Hall, Martin J R; Schwyn, Daniel A; Wicklein, Martina

2012-07-10

We demonstrate how micro-computed tomography (micro-CT) can be a powerful tool for describing internal and external morphological changes in Calliphora vicina (Diptera: Calliphoridae) during metamorphosis. Pupae were sampled during the 1st, 2nd, 3rd and 4th quarter of development after the onset of pupariation at 23 °C, and placed directly into 80% ethanol for preservation. In order to find the optimal contrast, four batches of pupae were treated differently: batch one was stained in 0.5M aqueous iodine for 1 day; two for 7 days; three was tagged with a radiopaque dye; four was left unstained (control). Pupae stained for 7d in iodine resulted in the best contrast micro-CT scans. The scans were of sufficiently high spatial resolution (17.2 μm) to visualise the internal morphology of developing pharate adults at all four ages. A combination of external and internal morphological characters was shown to have the potential to estimate the age of blowfly pupae with a higher degree of accuracy and precision than using external morphological characters alone. Age specific developmental characters are described. The technique could be used as a measure to estimate a minimum post-mortem interval in cases of suspicious death where pupae are the oldest stages of insect evidence collected. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

Gas-exchange patterns of Mediterranean fruit fly Pupae (Diptera: Tephritidae): A tool to forecast developmental stage

International Nuclear Information System (INIS)

Nestel, D.; Nemny-Lavy, E.; Alchanatis, V.

2007-01-01

The pattern of gas-exchange (CO 2 emission) was investigated for developing Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann) pupae incubated at different temperatures. This study was undertaken to explore the usefulness of gas-exchange systems in the determination of physiological age in developing pupae that are mass produced for sterile insect technique projects. The rate of CO 2 emission was measured in a closed flow-through system connected to commercial infrared gas analysis equipment. Metabolic activity (rate of CO 2 emission) was related to pupal eye-color, which is the current technique used to determine physiological age. Eye-color was characterized digitally with 3 variables (Hue, Saturation and Intensity), and color separated by discriminant analysis. The rate of CO 2 emission throughout pupal development followed a U-shape, with high levels of emission during pupariation, pupal transformation and final pharate adult stages. Temperature affected the development time of pupae, but not the basic CO 2 emission patterns during development. In all temperatures, rates of CO 2 emission 1 and 2 d before adult emergence were very similar. After mid larval-adult transition (e.g., phanerocephalic pupa), digital eye-color was significantly correlated with CO 2 emission. Results support the suggestion that gas-exchange should be explored further as a system to determine pupal physiological age in mass production of fruit flies. (author) [es

Cap-n-Collar Promotes Tissue Regeneration by Regulating ROS and JNK Signaling in the Drosophila melanogaster Wing Imaginal Disc.

Science.gov (United States)

Brock, Amanda R; Seto, Mabel; Smith-Bolton, Rachel K

2017-07-01

Regeneration is a complex process that requires an organism to recognize and repair tissue damage, as well as grow and pattern new tissue. Here, we describe a genetic screen to identify novel regulators of regeneration. We ablated the Drosophila melanogaster larval wing primordium by inducing apoptosis in a spatially and temporally controlled manner and allowed the tissue to regenerate and repattern. To identify genes that regulate regeneration, we carried out a dominant-modifier screen by assessing the amount and quality of regeneration in adult wings heterozygous for isogenic deficiencies. We have identified 31 regions on the right arm of the third chromosome that modify the regenerative response. Interestingly, we observed several distinct phenotypes: mutants that regenerated poorly, mutants that regenerated faster or better than wild-type, and mutants that regenerated imperfectly and had patterning defects. We mapped one deficiency region to cap-n-collar ( cnc ), the Drosophila Nrf2 ortholog, which is required for regeneration. Cnc regulates reactive oxygen species levels in the regenerating epithelium, and affects c-Jun N-terminal protein kinase (JNK) signaling, growth, debris localization, and pupariation timing. Here, we present the results of our screen and propose a model wherein Cnc regulates regeneration by maintaining an optimal level of reactive oxygen species to promote JNK signaling. Copyright © 2017 by the Genetics Society of America.

Analysis of synaptic growth and function in Drosophila with an extended larval stage.

Science.gov (United States)

Miller, Daniel L; Ballard, Shannon L; Ganetzky, Barry

2012-10-03

The Drosophila larval neuromuscular junction (NMJ) is a powerful system for the genetic and molecular analysis of neuronal excitability, synaptic transmission, and synaptic development. However, its use for studying age-dependent processes, such as maintenance of neuronal viability and synaptic stability, are temporally limited by the onset of pupariation and metamorphosis. Here we characterize larval NMJ growth, growth regulation, structure, and function in a developmental variant with an extended third instar (ETI). RNAi-knockdown of the prothoracicotropic hormone receptor, torso, in the ring gland of developing larvae leaves the timing of first and second instar molts largely unchanged, but triples duration of the third instar from 3 to 9.5 d (McBrayer et al., 2007; Rewitz et al., 2009). During this ETI period, NMJs undergo additional growth (adding >50 boutons/NMJ), and this growth remains under the control of the canonical regulators Highwire and the TGFβ/BMP pathway. NMJ growth during the ETI period occurs via addition of new branches, satellite boutons, and interstitial boutons, and continues even after muscle growth levels off. Throughout the ETI, organization of synapses and active zones remains normal, and synaptic transmission is unchanged. These results establish the ETI larval system as a viable model for studying motor neuron diseases and for investigating time-dependent effects of perturbations that impair mechanisms of neuroprotection, synaptic maintenance, and response to neural injury.

Timed Knickkopf function is essential for wing cuticle formation in Drosophila melanogaster.

Science.gov (United States)

Li, Kaixia; Zhang, Xubo; Zuo, Ying; Liu, Weimin; Zhang, Jianzhen; Moussian, Bernard

2017-10-01

The insect cuticle is an extracellular matrix that consists of the polysaccharide chitin, proteins, lipids and organic molecules that are arranged in distinct horizontal layers. In Drosophila melanogaster, these layers are not formed sequentially, but, at least partially, at the same time. Timing of the underlying molecular mechanisms is conceivably crucial for cuticle formation. To study this issue, we determined the time period during which the function of Knickkopf (Knk), a key factor of chitin organization, is required for wing cuticle differentiation in D. melanogaster. Although knk is expressed throughout metamorphosis, we demonstrate that its expression 30 h prior and 48 h after pupariation is essential for correct wing cuticle formation. In other words, expression beyond this period is futile. Importantly, manipulation of Knk expression during this time causes wing bending suggesting an effect of Knk amounts on the physical properties of the wing cuticle. Manipulation of Knk expression also interferes with the structure and function of the cuticle surface. First, we show that the shape of surface nano-structures depends on the expression levels of knk. Second, we find that cuticle impermeability is compromised in wings with reduced knk expression. In summary, despite the extended supply of Knk during metamorphosis, controlled amounts of Knk are important for correct wing cuticle differentiation and function in a concise period of time. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

Science.gov (United States)

Salazar-Souza, Mônica; Couri, Márcia S; Aguiar, Valeria M

2018-04-12

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.

Disruption of pupariation and eclosion behavior in the flesh fly, Sarcophaga bullata Parker (Diptera: Sarcophagidae), by venom form the ectoparasitic wasp Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae)

Czech Academy of Sciences Publication Activity Database

Rivers, D. B.; Žďárek, Jan; Denlinger, D. L.

2004-01-01

Roč. 57, - (2004), s. 78-91 ISSN 0739-4462 R&D Projects: GA ČR GA522/01/0501 Grant - others:USDA-NRI(US) 98-35302-6659; USDA-NRICGP(US) 2001-1005 Institutional research plan: CEZ:AV0Z4055905 Keywords : ectoparasitoid * ecdysone * fly development Subject RIV: CC - Organic Chemistry Impact factor: 1.173, year: 2004

Effect of morphine on the growth rate of Calliphora stygia (Fabricius) (Diptera: Calliphoridae) and possible implications for forensic entomology.

Science.gov (United States)

George, Kelly A; Archer, Melanie S; Green, Lauren M; Conlan, Xavier A; Toop, Tes

2009-12-15

Insect specimens collected from decomposing bodies enable forensic entomologists to estimate the minimum post-mortem interval (PMI). Drugs and toxins within a corpse may affect the development rate of insects that feed on them and it is vital to quantify these effects to accurately calculate minimum PMI. This study investigated the effects of morphine on growth rates of the native Australian blowfly, Calliphora stygia (Fabricius) (Diptera: Calliphoridae). Several morphine concentrations were incorporated into pet mince to simulate post-mortem concentrations in morphine, codeine and/or heroin-dosed corpses. There were four treatments for feeding larvae; T 1: control (no morphine); T 2: 2 microg/g morphine; T 3: 10 microg/g morphine; and T 4: 20 microg/g morphine. Ten replicates of 50 larvae were grown at 22 degrees C for each treatment and their development was compared at four comparison intervals; CI 1: 4-day-old larvae; CI 2: 7-day-old larvae; CI 3: pupae; and CI 4: adults. Length and width were measured for larvae and pupae, and costae and tibiae were measured for adults. Additionally, day of pupariation, day of adult eclosion, and survivorship were calculated for each replicate. The continued presence of morphine in meat was qualitatively verified using high-performance liquid chromatography with acidic potassium permanganate chemiluminescence detection. Growth rates of C. stygia fed on morphine-spiked mince did not differ significantly from those fed on control mince for any comparison interval or parameter measured. This suggests that C. stygia is a reliable model to use to accurately age a corpse containing morphine at any of the concentrations investigated.

Nonconsumptive Effects of Predatory Chrysomya rufifacies (Diptera: Calliphoridae) Larval Cues on Larval Cochliomyia macellaria (Diptera: Calliphoridae) Growth and Development.

Science.gov (United States)

Flores, Micah; Crippen, Tawni L; Longnecker, Michael; Tomberlin, Jeffery K

2017-09-01

Forensic entomologists often rely on development data associated with a given species to estimate when it colonized human or other vertebrate remains. In most instances, these development studies are based on single species reared in isolation in the laboratory. This study examined the impact of excretions and secretions (ES) associated with third-instar Chrysomya rufifacies (Macquart), a predator, on the development of its prey, Cochliomyia macellaria (F.). Not surprisingly, Ch. rufifacies ES did not impact the development of first- or second-instar C. macellaria, which are typically not preyed on by Ch. rufifacies. However, development of third-instar C. macellaria, which do experience predation, was impacted. First, larvae were longer than those in the control (deionized water, dH2O). Filtering the ES and removing the associated bacteria and byproducts >0.2 µm dampened the previous impact observed by the unfiltered ES on third-instar C. macellaria. Second, third-instar C. macellaria treated with unfiltered ES completed pupariation 8 h quicker than the controls. Filtering the ES lessened this effect by 50%. And finally, third-instar C. macellaria treated with filtered or unfiltered Ch. rufifacies ES reached adulthood ∼5 h faster than controls treated with dH2O. In summary, these data have large ramifications for forensic entomology, as multiple species being present on decomposing remains is not uncommon. Understanding the impact of associated ES produced by interspecific cohorts on associated development could lead to more precise estimates of the minimum postmortem interval for forensic investigation of decomposing remains. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Sterility induction in tsetse

Energy Technology Data Exchange (ETDEWEB)

Curtis, C F [London School of Hygiene and Tropical Medicine (UK); Langley, P A [Bristol Univ. (UK)

1982-01-01

The first laboratory and field experiments on genetic control were with tsetse flies, and they made use of the sterility in crosses and hybrids between closely related species of the Glossina morsitans complex. Backcrosses indicate that there are two separate sterility mechanisms involved: (a) genetic incompatibility between a mother and the products of alien genes in the embryo or larva in the uterus; (b) inability of hybrid males to inseminate due to incompatibility of their X-chromosomes with an alien Y or autosomes. The two largest sterile male release programmes have been in Tanzania and Upper Volta, and have used irradiation at the pupal or adult stage, respectively, for the production of sterility. Male tsetse are remarkably resistant to radiosterilization and, with the doses required to induce dominant lethals in more than 95% of sperms, premature senescence and lethargic behaviour of the males tends to result. With G. morsitans irradiated at the puparial stage these effects can be alleviated by the use of a nitrogen atmosphere during irradiation. If the puparia are then transferred to air at 11/sup 0/C for transport to the release site, immediate emergence occurs on re-warming after arrival. This advantageous procedure was used for the releases in Tanzania. In addition to dominant lethals, irradiation also produces chromosome translocations which cause inherited partial sterility. A homozygous translocation line was selected but this example did not have sufficient fitness to be used in a mass rearing programme. Chemosterilants can be applied by pupal dipping, adult contact with deposits or in aerosols. Studies are now in progress on their use in association with odour-baited traps or pheromone-baited decoys as a means of sterilizing the wild population and thus avoiding the costly and difficult process of mass rearing tsetse.

Developmental rates of immatures of three Chrysomya species (Diptera: Calliphoridae) under the effect of methylphenidate hydrochloride, phenobarbital, and methylphenidate hydrochloride associated with phenobarbital.

Science.gov (United States)

Rezende, Fábio; Alonso, Marcela A; Souza, Carina M; Thyssen, Patrícia J; Linhares, Arício X

2014-05-01

Entomotoxicology is focused on obtaining data on necrophagous entomofauna, for criminal investigations purposes. This study aimed to evaluate the effect of different concentrations of methylphenidate hydrochloride, phenobarbital, and their association on the developmental rate, larval and pupal survivorship, and the interval of emergence of adults of Chrysomya albiceps (Wiedemann), Chrysomya megacephala (Fabricius), and Chrysomya putoria (Wiedemann) (Diptera: Calliphoridae). Considering the therapeutic dose (TD) of methylphenidate hydrochloride (0.29 mg/Kg), the concentrations tested were 10× TD, 50× TD, and 100× TD. For phenobarbital, the concentrations used were 1× TD (=150 mg/Kg), 3.3× TD, and 6.7× TD. For the association of the drugs, the combinations used were 10× TD-methylphenidate hydrochloride plus 1× TD-phenobarbital, 50× TD-methylphenidate hydrochloride plus 3.3× TD-phenobarbital, and 100× TD-methylphenidate hydrochloride plus 6.7× TD-phenobarbital. The control group, without addition of drug, was maintained under the same conditions of temperature (25 ± 1 °C), humidity (70 ± 10%), and photoperiod (12 h). Specimens of each group were weighed every 12 h until pupariation. The developmental rate of the three Chrysomya species immatures was monitored. For C. albiceps the developmental time was delayed in 24 h for methylphenidate hydrochloride group and in 12 h for the phenobarbital and the drugs association groups. The effect was observed only at specific ages for C. megacephala, without altering the developmental time. For C. putoria, the developmental time was delayed in 12 h for methylphenidate hydrochloride group and in 24 h for the phenobarbital and the drugs association groups. The emergence interval was similar among all experimental groups, but larval and pupal viabilities were affected in different ways.

Induction of the early-late Ddc gene during Drosophila metamorphosis by the ecdysone receptor.

Science.gov (United States)

Chen, Li; Reece, Christian; O'Keefe, Sandra L; Hawryluk, Gregory W L; Engstrom, Monica M; Hodgetts, Ross B

2002-06-01

During Drosophila metamorphosis, the 'early-late' genes constitute a unique class regulated by the steroid hormone 20-hydroxyecdysone. Their induction is comprised of both a primary and a secondary response to ecdysone. Previous work has suggested that the epidermal expression of the dopa decarboxylase gene (Ddc) is likely that of a typical early-late gene. Accumulation of the Ddc transcript is rapidly initiated in the absence of protein synthesis, which implies that the ecdysone receptor plays a direct role in induction. However, full Ddc expression requires the participation of one of the transcription factors encoded by the Broad-Complex. In this paper, we characterize an ecdysone response element (EcRE) that contributes to the primary response. Using gel mobility shift assays and transgenic assays, we identified a single functional EcRE, located at position -97 to -83 bp relative to the transcription initiation site. This is the first report of an EcRE associated with an early-late gene in Drosophila. Competition experiments indicated that the affinity of the Ddc EcRE for the ecdysone receptor complex was at least four-fold less than that of the canonical EcRE of the hsp27 gene. Using in vitro mutagenesis, we determined that the reduced affinity of the EcRE resided at two positions where the nucleotides differed from those found in the canonical sequence. The ecdysone receptor, acting through this EcRE, releases Ddc from a silencing mechanism, whose cis-acting domain we have mapped to the 5'-upstream region between -2067 and -1427 bp. Deletion of this repressive element resulted in precocious expression of Ddc in both epidermis and imaginal discs. Thus, epidermal Ddc induction at pupariation is under the control of an extended genomic region that contains both positive and negative regulatory elements. Copyright 2002 Elsevier Science Ireland Ltd.

Electron beam irradiation induces abnormal development and the stabilization of p53 protein of American serpentine leafminer, Liriomyza trifolii (Burgess)

Energy Technology Data Exchange (ETDEWEB)

Koo, Hyun-Na; Yun, Seung-Hwan; Yoon, Changmann [Department of Plant Medicine, College of Agriculture, Life and Environment Sciences, Chungbuk National University, Cheongju 361-763 (Korea, Republic of); Kim, Gil-Hah, E-mail: khkim@chungbuk.ac.kr [Department of Plant Medicine, College of Agriculture, Life and Environment Sciences, Chungbuk National University, Cheongju 361-763 (Korea, Republic of)

2012-01-15

The American serpentine leafminer fly, Liriomyza trifolii (Burgess), is one of the most destructive polyphagous pests worldwide. In this study, we determined electron beam doses for inhibition of normal development of the leaf miner and investigated the effect of electron beam irradiation on DNA damage and p53 stability. Eggs (0-24 h old), larvae (2nd instar), puparia (0-24 h old after pupariation) and adults (24 h after emergence) were irradiated with increasing doses of electron beam irradiation (six levels between 30 and 200 Gy). At 150 Gy, the number of adults that developed from irradiated eggs, larvae and puparia was lower than in the untreated control. Fecundity and egg hatchability decreased depending on the doses applied. Reciprocal crosses between irradiated and unirradiated flies demonstrated that males were more radiotolerant than females. Adult longevity was not affected in all stages. The levels of DNA damage in L. trifolii adults were evaluated using the alkaline comet assay. Our results indicate that electron beam irradiation increased levels of DNA damage in a dose-dependent manner. Moreover, low doses of electron beam irradiation led to the rapid appearance of p53 protein within 6 h; however, it decreased after exposure to high doses (150 Gy and 200 Gy). These results suggest that electron beam irradiation induced not only abnormal development and reproduction but also p53 stability caused by DNA damage in L. trifolii. We conclude that a minimum dose of 150 Gy should be sufficient for female sterilization of L. trifolii. - Highlights: > Electron beam irradiation inhibited normal development of the leaf miner. > Electron beam irradiation inhibited normal reproduction of the leaf miner. > Electron beam irradiation increased levels of DNA damage. > Electron beam irradiation induced p53 stability.

Parturition in Tsetse Flies: Endocrine Control

Energy Technology Data Exchange (ETDEWEB)

Zd' arek, J; Cvacka, J; Sanda, M [Institute of Organic Chemistry and Biochemistry, AV CR, Prague (Czech Republic); Takac, P; Keszeliova, D; Simo, L; Roller, L [Institute of Zoology, SAV, Bratislava (Slovakia)

2012-07-15

A problem in tsetse mass rearing facilities is the increased incidence of abortions of underdeveloped larvae or pupariation of larvae within the mother's uterus. We analysed the problem by investigating neural, hormonal and environmental factors controlling parturition. Input from the mother's brain is essential for normal parturition, since a female whose brain is disconnected by ligation fails to deposit her larva. The expulsion of a larva is stimulated by a putative parturition hormone present within the female's uterus. The hormone also elicits abortion when injected into neck ligated females at earlier stages of pregnancy. This report describes attempts to reveal the chemical nature of this hormone by purification of extracts of uteri of Glossina females and identification of behaviourally active fractions using a MALDI-MS instrument. Genomic (BLAST) analysis of the identified sequences did not reveal a significant match with any protein with bioactive properties in other species. However, similarity with various enzymes or structural proteins (and hypothetical proteins) was detected occasionally. In the Glossina genomic and cDNA databases no nucleotide sequence corresponding to the deduced AA sequences was found. Perhaps the deduced sequences are too short to obtain more significant hits both in protein and nucleotide databases. We also made investigations to elucidate environmental influences and physiological mechanisms associated with tsetse parturition. We found that the circadian rhythm of parturition of flies kept in Bratislava (G. m. morsitans, G. f. fuscipes and G. pallidipes) is less pronounced than under natural conditions. The loss of synchrony in the laboratory may have three possible causes: (i) genetic - absence of selective pressure, (ii) environmental - low intensity or absence of an entraining light or temperature stimulus, and (iii) physiological - impaired sensitivity to olfactory stimulation by a hypothetical 'oviposition' pheromone that