In search of the spectral composition of an effective light trap for the mushroom pest Lycoriella ingenua (Diptera: Sciaridae).

Certain fungus gnats, like Lycoriella ingenua are notorious pests in agriculture, especially in mushroom production. While larvae cause mainly direct crop damage, adults are vectors of several dangerous fungal pathogens. To promote the development of pesticide-free management methods, such as light trapping, we measured the spectral sensitivity of L. ingenua compound eyes with electroretinography and performed two different behavioural experiments to reveal the wavelength dependence of phototaxis in this species. The spectral sensitivity of the compound eyes is bimodal with peaks at 370 nm (UV) and 526 nm (green). Behavioural experiments showed that attraction to light as a function of wavelength depends on light intensity. In our first experiment, where the minimal photon flux (105-109 photons/cm2/s) needed for eliciting a phototactic response was determined wavelength by wavelength, phototaxis was strongest in the green spectral range (~526 nm). In the other behavioural experiment, where wavelength preference was tested under a higher but constant light intensity (~1013 photons/cm2/s), the highest attraction was elicited by UV wavelengths (398 nm). Our results suggest that both UV and green are important spectral regions for L. ingenua thus we recommend to use both UV (~370-398 nm) and green (~526 nm) for trapping these insects.

RNA-Seq analysis of the blue light-emitting Orfelia fultoni (Diptera: Keroplatidae) suggest photoecological adaptations at the molecular level.

Bioluminescence in Diptera is found in the Keroplatidae family, within Arachnocampininae and Keroplatinae subfamilies, with reported occurrences in Oceania, Eurasia, and Americas. Larvae of Orfelia fultoni, which inhabit stream banks in the Appalachian Mountains, emit the bluest bioluminescence among insects, using it for prey attraction, similarly to Arachnocampa spp. Although bioluminescence has a similar prey attraction function, the systems of Arachonocampininae and Keroplatinae subfamilies are morphologically/biochemically distinct, indicating different evolutionary origins. To identify the possible coding genes associated with physiological control, ecological adaptations, and origin/evolution of bioluminescence in the Keroplatinae subfamily, we performed the RNA-Seq analysis of O. fultoni larvae during day and night and compared it with the transcriptomes of Arachnocampa luminosa, and reanalyzed the previously published proteomic data of O. fultoni against the RNA-Seq dataset. The abundance of chaperones/heat-shock and hexamerin gene products at night and in luciferase enriched fractions supports their possible association and participation in bioluminescence. The low diversity of copies/families of opsins indicate a simpler visual system in O. fultoni. Noteworthy, gene products associated with silk protein biosynthesis in Orfelia were more similar to Lepidoptera than to the Arachnocampa, indicating that, similarly to the bioluminescent systems, at some point, the biochemical apparatus for web construction may have evolved independently in Orfelia and Arachnocampa.

Impact of artificial light intensity on nocturnal insect diversity in urban and rural areas of the Asir province, Saudi Arabia.

Continuous urban developments have resulted in increased demand for street furniture, one of which is street light columns. Artificial light at night (ALAN) pose significant impacts on insect diversity in urban and rural areas. The ALAN is a significant driver of decline in insect diversity. This study evaluated the impact of light intensity and sky quality at night on insect diversity in rural and urban areas of the Asir province, Saudi Arabia. Insect traps were installed in both areas during night. Light intensity of nearby road lamps was measured using light meter, while sky quality was measured using sky quality meter. Rural areas exhibited low light intensity (10.33 flux/f.candle) and good sky quality (18.80 magnitude/arcsec2). Urban areas exhibited intense light (89.33 flux/f.candle) and poor sky quality (15.49 magnitude/arcsec2). Higher insect diversity was recorded for rural areas where insects belonging to seven orders (i.e., Diptera, Lepidoptera, Hemiptera, Hymenoptera, Coleoptera, Neuroptera, and Dermaptera) were collected. However, insects of four orders (i.e., Diptera, Lepidoptera, Hemiptera, and Neuroptera) were found in urban areas indicating low diversity. Lepidopteran insects were frequently recorded from rural areas indicating they are attracted to artificial light. It is concluded that excessive ALAN and poor sky quality at night disrupt insect biodiversity. Therefore, ALAN and sky quality must be considered responsible for decline in insect biodiversity along with other known factors.

Mating success of the black soldier fly, Hermetia illucens (Diptera: Stratiomyidae), under four artificial light sources.

Larvae of the black soldier fly (BSF) Hermetia illucens (L.) (Diptera: Stratiomyidae) are promising organisms to be used for organic waste bioconversion. Breeding BSF indoors has been suggested as a cost-effective approach for countries with long winters or low sunlight levels through the year. As the BSF mating is visually mediated, artificial illumination conditions are critical to a successful indoor breeding system. In this study, we tested four different types of artificial light sources for their effects on BSF mating success. They included: (1) a halogen lamp; (2) a combination of a white light-emitting diode (LED) lamp and a fluorescent ultra violet lamp; (3) a metal halide lamp, and (4) a specially designed light-emitting diode (BSFLED) lamp, whose design was based on the specific BSF adult visual spectral sensitivity. We determined the spectra of four artificial light sources, compared their spectral composition in relation to the BSF-visible spectrum, and compared their effects on the mating success of two different BSF colonies. BSFLED was the most energy efficient light source in spectral composition and led to the highest mating success in terms of the percentage of inseminated females and fertile clutches. Thus, BSFLED is the most suitable light source tested in our experiment for breeding BSF indoors. The colony effect and possible light flickering effect on BSF mating success were also detected. The implications of these findings are discussed.

Hessian Fly (Diptera: Cecidomyiidae) Attraction to Different Wavelengths and Intensities of Light-Emitting Diodes in the Laboratory.

Monitoring of Hessian fly, Mayetiola destructor (Say) (Diptera: Cecidomyiidae), populations is important for targeted management methods. Also, effectiveness of monitoring efforts is critical to surveillance efforts in regions of the world without this pest. Current Hessian fly monitoring traps rely purely on a single attractant, the female sex pheromone, which is ineffective for monitoring females in the population. Our objective was to examine another attractant targeting vision of both males and females. To do this, we evaluated emitted light of various wavelengths and intensities produced by light-emitting diodes (LEDs). Hessian flies were released in the center of a four-leaf clover-shaped arena, which contained an LED within each collection cup at the apex of each of the four cloverleaves. Initially, we examined a range of colors, including red, amber, green, and blue, with wavelengths of 624, 590, 527, and 472 nm, respectively. Both sexes of Hessian fly preferred green LEDs; therefore, we examined specific wavelengths within the green spectrum (502, 525, and 565 nm), and varied light intensities (4, 8, 12, and 16 W/m2). Specifically, females preferred wavelengths in the lower region of the green spectrum (502 and 525 nm), and both sexes preferred high intensity light (16 W/m2). This is the first report of Hessian fly attraction to select emitted wavelengths and intensities from LEDs under controlled conditions. Leveraging these results into new trap designs will add a second sensory modality to the existing trap; however, future studies are needed to assess attraction to LED traps under field conditions.

Cloning and functional characterizations of an apoptogenic Hid gene in the Scuttle Fly, Megaselia scalaris (Diptera; Phoridae).

Although the mechanisms of apoptotic cell death have been well studied in the fruit fly, Drosophila melanogaster, it is unclear whether such mechanisms are conserved in other distantly related species. Using degenerate primers and PCR, we cloned a proapoptotic gene homologous to Head involution defective (Hid) from the Scuttle fly, Megaselia scalaris (MsHid). MsHid cDNA encodes a 197-amino acid-long polypeptide, which so far is the smallest HID protein. PCR analyses revealed that the MsHid gene consists of four exons and three introns. Ectopic expression of MsHid in various peptidergic neurons and non-neuronal tissues in Drosophila effectively induced apoptosis of these cells. However, deletion of either conserved domain, N-terminal IBM or C-terminal MTS, abolished the apoptogenic activity of MsHID, indicating that these two domains are indispensable. Expression of MsHid was found in all life stages, but more prominently in embryos and pupae. MsHid is actively expressed in the central nervous system (CNS), indicating its important role in CNS development. Together MsHID is likely to be an important cell death inducer during embryonic and post-embryonic development in this species. In addition, we found 2-fold induction of MsHid expression in UV-irradiated embryos, indicating a possible role for MsHid in UV-induced apoptosis.

Comparison of different light sources for trapping Culicoides biting midges, mosquitoes and other dipterans.

The response of Culicoides biting midges, mosquitoes and other dipterans to different wavelengths was evaluated in a farm meadow in northern Spain. A total of 9449 specimens of 23 species of Culicoides, 5495 other ceratopogonids (non-biting midges), 602 culicids and 12428 other mixed dipterans were captured. Centers for Disease Control and Prevention (CDC) suction light traps fitted with five light emitting diodes (LEDs) (white, green, red, blue, ultraviolet) were run for 15 consecutive nights. Significantly more Culicoides were collected in those traps fitted with green, blue or ultraviolet (UV) lights than in red and white-baited LED traps for the most abundant species captured: C. punctatus (37.5%), C. cataneii (26.5%) and C. obsoletus/C. scoticus (20.4%). Similar results were obtained for non-Culicoides ceratopogonids, mosquitoes and other mixed dipterans. Wavelengths in green (570nm) resulted effective for targeting some Culicoides species, culicids and other midges. In a second trial, the effectiveness of 4-W white and UV tubes was compared to traps fitted with UV LED and a standard incandescent light bulb. More specimens of all taxa were collected with fluorescent black light (UV) traps than with the other light sources, except culicids, which were recovered in high numbers from fluorescent white light traps.

How photoperiods affect the immature development of forensically important blowfly species Chrysomya albiceps (Calliphoridae).

The purpose of this study was to verify the influence of different photoperiods on larval body weight, post-embryonic development, and viability of Chrysomya albiceps. The bioassays were performed in acclimatized chamber at 27 ± 1 °C and 60 ± 10 % relative humidity regulated by three different light and dark cycles for a period of 24 h-12:12, 24:00, and 00:24. Four replications with 50 newly hatched larvae each were used per photoperiod. The larvae were placed in container containing bovine ground meat (50 g). When mature larvae spontaneously abandoned the diet, they were individually weighed and separated into glass tubes until emergence. The larvae that did not abandon the diet were not weighed. The larvae from 24 h of photophase did not abandon the diet and pupated inside, so these larvae were not weighed and the larval and pupal developments were not registered. The mean of larval body weight did not vary between the 24 h scotophase and 12 h photophase. The mean duration of larval developmental time varied significantly when comparing the 24 h scotophase and the 12 h photophase, and the pupal developmental time also varied for the same photoperiods; in both stages, the duration of development was faster in the 24 h scotophase. The developmental time of neo-larvae to adult was verified in all photoperiods and gradually increased as photophase increased. There was a trend to augment of viability with the reduction of photophase length.

Photodynamic treatment of Chaoborus crystallinus larvae with chlorophyllin induces necrosis and apoptosis.

Chlorophyllin kills mosquito larvae (Culex, Aedes) in the aquatic habitat at low concentrations via photodynamic reactions under irradiation. The effects of chlorophyllin were investigated at the cellular level using the transparent larvae of Chaoborus crystallinus as a model system. Their transparency enabled in situ fluorescence investigation, showing that chlorophyllin accumulates in the intestine of the larvae. Uptake of chlorophyllin at room temperature took about 2 h. The fluorescence signal peaked after 5 h of incubation. Chlorophyllin accumulates up to about 15 ng per larvae. The intestine of treated larvae was dissected and stained with several dyes (acridine orange, Hoechst 33342 and propidium iodide). Apoptosis and necrosis increased with higher concentrations of chlorophyllin (to a smaller extent in dark controls) and were elevated in irradiated samples. Single cells from treated larvae were isolated and subjected to Annexin V flow cytometry. The fraction of apoptotic and necrotic cells increased significantly at a high chlorophyllin concentration (21.4 mg L(-1)) and under intensive irradiation. The activity of caspases-3, -8 and -9 as well as Bcl-2 and cytochrome c was investigated by means of western blot analysis. The data suggest a possible chlorophyllin concentration-dependent shift of the apoptotic pathway.

An artificial light source influences mating and oviposition of black soldier flies, Hermetia illucens.

Current methods for mass-rearing black soldier flies, Hermetia illucens (L.) (Diptera: Stratiomyidae), in the laboratory are dependent on sunlight. Quartz-iodine lamps and rare earth lamps were examined as artificial light sources for stimulating H. illucens to mate and lay eggs. Sunlight was used as the control. Adults in the quartz-iodine lamp treatment had a mating rate of 61% of those in the sunlight control. No mating occurred when the rare earth lamp was used as a substitute. Egg hatch for the quartz-iodine lamp and sunlight treatments occurred in approximately 4 days, and the hatch rate was similar between these two treatments. Larval and pupal development under these treatments required approximately 18 and 15 days at 28°C, respectively. Development of methods for mass rearing of H. illucens using artificial light will enable production of this fly throughout the year without investing in greenhouse space or requiring sunlight.

Clock genes period and timeless are rhythmically expressed in brains of newly hatched, photosensitive larvae of the fly, Sarcophaga crassipalpis.

While roles of the clock genes period (per) and timeless (tim) are relatively well understood in relation to circadian clocks, their potential roles in insect photoperiodism remain enigmatic. In this study, the expression of per and tim genes under two contrasting photoperiods is described in the central nervous system of photoperiodically sensitive, newly hatched first instar larvae of the flesh fly, Sarcophaga crassipalpis. Using qPCR, diel oscillations were observed in the mRNA levels of both genes under long-day (15 h light:9h dark, promotes direct development) and short-day conditions (11 h light:13 h dark, induces pupal diapause). Peak per and tim mRNA oscillations were closely associated with the light/dark transition. The conspicuous difference between the two photoperiodic conditions was that the sharp increase in per and tim mRNA abundance occurred during the light phase under long days but during the dark phase under short days. The diel oscillations were, at least in part, driven by an endogenous component, as demonstrated by transferring larvae to continuous darkness. The cells displaying Tim- and Per-like immunoreactivities (Tim- and Per-LIRs) were localized using anti-Drosophila-Per and anti-Chymomyza-Tim antibodies. Per-LIR and Tim-LIR co-localized in three groups of cells in each brain hemisphere. Two other groups, one in the brain hemispheres and the other in the fused ventral nerve ganglion, expressed only the Per-LIR.

Possible involvement of distinct photoreceptors in the photoperiodic induction of diapause in the flesh fly Sarcophaga similis.

Physiological characteristics of the photoreceptors involved in the photoperiodic induction of diapause were investigated in the flesh fly Sarcophaga similis. Both the early and late phases of scotophase were sensitive to light and a light pulse during each of these phases prevented diapause. Certain physiological differences between the phases were, nevertheless, detected. Compared with early scotophase, late scotophase required a light pulse with a long period and a large number of night interruption photoperiodic cycles in order to effectively prevent diapause. The diapause-averting effects of a light pulse during early scotophase were canceled by an additional long dark period, but those during late scotophase were not. Thus, the diapause-averting effects produced during early scotophase are different to those produced during late scotophase. The early scotophase was sensitive to light at wavelengths of 470nm or shorter, but not to light of 583nm or longer. In contrast, the late scotophase was sensitive to light of a broad range of wavelengths, ranging from 395 to 660nm. Furthermore, the early scotophase was considerably more sensitive to monochromatic light with low photon flux density than the late scotophase. These results suggest that different types of photoreceptor are involved in the photoperiodic response.

Actin and myosin inhibitors block elongation of kinetochore fibre stubs in metaphase crane-fly spermatocytes.

We used an ultraviolet microbeam to cut individual kinetochore spindle fibres in metaphase crane-fly spermatocytes. We then followed the growth of the "kinetochore stubs", the remnants of kinetochore fibres that remain attached to kinetochores. Kinetochore stubs elongate with constant velocity by adding tubulin subunits at the kinetochore, and thus elongation is related to tubulin flux in the kinetochore microtubules. Stub elongation was blocked by cytochalasin D and latrunculin A, actin inhibitors, and by butanedione monoxime, a myosin inhibitor. We conclude that actin and myosin are involved in generating elongation and thus in producing tubulin flux in kinetochore microtubules. We suggest that actin and myosin act in concert with a spindle matrix to propel kinetochore fibres poleward, thereby causing stub elongation and generating anaphase chromosome movement in nonirradiated cells.

Towards environmentally and human friendly insect pest control technologies: photosensitization of leafminer flies Liriomyza bryoniae.

Development of new, ecologically safe technologies to control insect pest populations is of great importance. Photoactive compounds usually used for photosensitization might be effective as pesticide agents, with low impact on the environment, being non-toxic and not mutagenic. Phosensitizer accumulates within the insect body and, following exposure to visible light, induces lethal photochemical reactions and death. The aim of this study is to evaluate the possible usage of several photosensitizers (acridine orange, aminolevulinic acid, hematoporphyrin dimethyl ether, methylene blue) as photopesticides to control population of polyphagous plant pest Liriomyza bryoniae (Kaltenbach, 1858) (Diptera, Agromyzidae). Fluorescence measurements of intact cooled insects indicate that insect feeding with bait containing HPde and sugar induces remarkable accumulation of this compound in the body of insect. This accumulation is strongly dependent on sex and feeding duration. The highest HPde amount in the body of insect was detected 16 h after feeding, whereas no significant photosensitizer amount was detected in the same insect following 48 h. Following irradiation with visible light results in fast death of L. bryoniae. Of importance to note that survival of insects after feeding and irradiation depends on sex: female insect died much faster than males.

Biological effects of anhydrobiosis in an African chironomid, Polypedilum vanderplanki on radiation tolerance.

PURPOSE: Anhydrobiotic organisms are known to have an extremely high tolerance against a range of stresses. However, the functional role of anhydrobiosis in radiation tolerance is poorly understood, especially in development following irradiation. The present study aims to evaluate effects of anhydrobiosis on radiation tolerance in an anhydrobiotic insect, Polypedilum vanderplanki. MATERIALS AND METHODS: Larval survival (48 h), anhydrobiotic ability, metamorphosis and reproduction after exposure to 1-9000 Gy of gamma-rays at the larval stage were compared between anhydrobiotic (dry) and normal (wet) phases. RESULTS: Wet larvae were killed in a dose-dependent manner at doses higher than 2000 Gy, and all died within 8 h after 4000 Gy exposure. In contrast, dry larvae survived even 5000 Gy, and some of them still tolerated 7000 Gy and were alive at 48 h after rehydration. Moreover, greater radiotolerance of dry larva, compared to wet ones, was demonstrated in terms of metamorphoses. However, anhydrobiosis did not protect against radiation damage in terms of producing viable offspring. CONCLUSION: These results indicate that anhydrobiosis enhances radiotolerance, resulting in increases of successful metamorphoses.

Sterilisation of Lucilia cuprina Wiedemann maggots used in therapy of intractable wounds.

Three new techniques of sterilising maggots of Lucilia cuprina for the purpose of debriding intractable wounds were studied. These techniques were utilisation of ultra-violet C (UVC) and maggot sterilisation with disinfectants. The status of sterility was checked on nutrient agar and blood agar and confirmed with staining. The indicators for the effectiveness of the methods were sterility and survival rate of the eggs or larvae. Egg sterilisation with UVC had the lowest hatching rate (16+/-0.00%) while egg sterilisation with disinfectants showed high hatching rate (36.67+/-4.41%) but low maggot survival rate (31.67+/-1.67%). Sterilisation of the maggots was the most suitable, since the survival rate was the highest (88.67+/-0.88%). Complete sterility was achieved in all cases, except that Proteus mirabilis was consistently found. However, the presence of this microorganism was considered beneficial.

SEM observation of the insect prepared by microwave irradiation.

The ptilinum of the fly and the compound eye are among the most fragile organs encountered during conventional procedures of morphological sample fixation. In order to identify a fixative suitable for preparing such samples for scanning electron microscopy, we examined various fixation conditions using microwave irradiation (MWI). The conditions examined were: (i) fixatives; (ii) temperature; (iii) concentration; (iv) duration; (v) dehydration; and (vi) substitution. The identified optimal conditions were 5% glutaraldehyde with MWI (350 W, 5 min). The MWI was continued until the maximal temperature of 75 degrees C was attained, followed by intermittent irradiation to maintain a temperature of 75 degrees C. After irradiation, the sample was left at room temperature for 24 h in the fixative and then dehydrated in increasing concentrations of ethanol. Each step in the ethanol series lasted for 24 h. The final absolute ethanol step included three solution changes, with each incubation lasting 1 h. A subsequent stepwise substitution of t-butyl alcohol for ethanol was conducted by reducing the ratio of 100% ethanol to t-butyl alcohol from 2:1 to 1:1 and then 1:2 (24 h each). The substitution was completed by three solution changes using 100% t-butyl alcohol, 30 min each. The best results were obtained by freeze-drying samples using t-butyl alcohol. The use of MWI improved fixative permeation, which occurred at a uniform rate throughout the sample. Comparison with temperature in a water bath at 75 degrees C indicated that the fixation effect of MWI was due to its heat generation in addition to some unknown mechanism.

Two bioluminescent diptera: the North American Orfelia fultoni and the Australian Arachnocampa flava. Similar niche, different bioluminescence systems.

Orfelia fultoni is the only bioluminescent dipteran (Mycetophilidae) found in North America. Its larvae live on stream banks in the Appalachian Mountains. Like their Australasian relative Arachnocampa spp., they build sticky webs to which their bioluminescence attracts flying prey. They bear two translucent lanterns at the extremities of the body, histologically distinct from the single caudal lantern of Arachnocampa spp., and emit the bluest bioluminescence recorded for luminescent insects (lambda(max) = 460 nm versus 484 nm from Arachnocampa). A preliminary characterization of these two bioluminescent systems indicates that they are markedly different. In Orfelia a luciferin-luciferase reaction was demonstrated by mixing a hot extract prepared with dithiothreitol (DTT) under argon with a crude cold extract. Bioluminescence is not activated by adenosine triphosphate (ATP) but is strongly stimulated by DTT and ascorbic acid. Using gel filtration, we isolated a luciferase fraction of approximately 140 kDa and an additional high molecular weight fraction (possibly a luciferin-binding protein) that activated bioluminescence in the presence of luciferase and DTT. The Arachnocampa luciferin-luciferase system involves a 36 kDa luciferase and a luciferin soluble in ethyl acetate under acidic conditions; the bioluminescence is activated by ATP but not by DTT. The present findings indicate that the bioluminescence of O. fultoni constitutes a novel bioluminescent system unrelated to that of Arachnocampa.

Female sensitivity to diet and irradiation treatments underlies sex-mortality differentials in the Mediterranean fruit fly.

Large-scale experiments on medflies that were subjected to sterilizing doses of ionizing radiation (plus intact controls) and maintained on either sugar-only or full, protein-enriched diets revealed that, whereas the mortality trajectories of both intact and irradiated male cohorts maintained on both diets are similar, the mortality patterns of females are highly variable. Mean mortality rates at 35 days in male cohorts ranged from 0.2 to 0.3 but in female cohorts ranged from 0.09 to 0.35, depending on treatment. The study reports three main influences: (a) qualitative differences exist in the sex-mortality response of medflies subjected to dietary manipulations and irradiation; (b) the female mortality response is linked to increased vulnerability due to the nutritional demands of reproduction; and (c) female sensitivity to environmental changes underlies the dynamics of the sex-mortality differential.

[Frequency of chromosome aberrations after irradiation of gametes of cabbage root fly (Delia brassicae Bouche) with various doses of X rays]. / Chastota khromosomnykh perestroek pri obluchenii polovykh kletok vesennei kapustnoi mukhi (Delia brassicae Bouche) raznymi dozami rentgenovskikh luchei.

Chromosome rearrangements were isolated via crossing of F1 offspring from X-irradiated male and female Delia brassicae with intact insects. An enhanced (more than 30%) rate of late embryonic lethality (LEL) in clutches was the primary criterion of rearrangements. When males alone were irradiated at 2.5, 5, and 15 Gy, a total of 7.8, 9.6, and 23.6% of their offspring, respectively, inherited semisterility. After irradiation of females alone, the LEL rate in F1 was insignificant. Hybridization analysis of F2 revealed 18 lines with autosomal inheritance of semisterility. Fourteen lines (43.7%) exhibited an increased rate of LEL linked to the male sex.