Humoral factor activating the Sarcophaga lectin gene in cultured fat body

On culture of the fat body of Sarcophaga peregrina (flesh fly) larvae in modified Grace's medium, the Sarcophaga lectin gene was activated only when the medium was supplemented with larval hemolymph. The expressions of the genes for the storage protein and the sarcocystatin A were not affected by supplementing the medium with the hemolymph. Thus the hemolymph contained a factor that specifically activated the Sarcophaga lectin gene. This factor seemed to be a heat-stable, low molecular weight compound that was probably not a peptide, and to activate genes in the fat body for defense proteins that are known to be expressed in response to injury of Sarcophaga larvae.     

Genomic organization in the flesh fly Sarcophaga bullata

The genome of the flesh fly Sarcophaga bullata has been characterized both cytologically and biochemically. S. bullata has a haploid DNA level of 0.61 picograms which is five times larger than the haploid genome size of Drosophila melanogaster. Reassociation kinetics of Sarcophaga DNA shows that its sequence organization is very similar to that of D. melanogaster in having a very large proportion of single copy DNA (81%) and only small amounts of highly and moderately repetitive DNA (9% and 6%, respectively). cRNAs from all three sequence classes were prepared and their cytological distributions on diploid and polytene cells determined by in situ hybridization. The cytological distribution of the highly repetitive probe was found to be restricted to the centromeric heterochromatin of two of the five autosomes and this sequence class was also found to be markedly underreplicated in polytene foot-pad cells. No highly repetitive DNA was localized on either of the sex chromosomes, but only on the two large centromeric regions of chromosomes C and E. Moderately repetitive DNA was found uniformly distributed on all of the autosomes in both testis and polytene foot-pad squashes. As in the case of the highly repetitive sequence probe, no moderately repetitive DNA was detected on either the X or Y chromosomes. Moderately repetitive DNA in Sarcophaga was also shown to have the Drosophila type pattern of sequence interspersion with a moderately repetitive element of 5,000 nucleotides adjacent to a unique element of greater than 10,000 nucleotides. The Sarcophaga genome is the largest for which this type of interspersion has so far been demonstrated.     

What determines the number of ovarioles in a fly ovary?

The relation between the size of a fly and the number of ovarioles in its ovary was investigated in Phormia regina and Sarcophaga bullata. Small flies of varying size were produced by taking larvae prematurely off the food. The smallest flies thus obtained were derived from larvae only $\text{1}\text{8}$ of the weight of a normal larva. The number of ovarioles in an ovary is directly proportional to the size of the fly and, in the extreme case, is about $\text{1}\text{5}$ the normal number in Sarcophaga and about $\text{1}\text{3}$ in Phormia. Larvae prematurely taken off the food, but fed again after starving for several days, grow to normal or almost normal size and develop ovaries with about the normal number of ovarioles. Small or re-fed Sarcophaga do not show any changes in the anatomy of individual ovarioles but in Phormia disorders in ovariole development and a consequent reduction of fertility are frequent. The number of ovarioles remains identical from the early pupal stage all through the development of the pharate fly and then through ovarian development in the adult fly: it is determined by the size of the larva when it was taken off the food. This shows that it is not lack of space in a small adult fly abdomen which determines the number nor the occurrence of degenerative processes during ovarian development.     

Towards a phylogeny of the flesh flies (Diptera: Sarcophagidae): morphology and phylogenetic implications of the acrophallus in the subfamily Sarcophaginae

The morphology of the acrophallus, the distal portion of the male phallus carrying the phallotreme, was studied in 72 exemplar species representing 56 genera and subgenera of the flesh fly subfamily Sarcophaginae. For 42 of those species, scanning electron microscopy was used to clarify the phallic morphology. Terms used to describe the male genitalia were updated based on new interpretations of homology. Male genitalic characters, combined with other morphological characters of adult males and females and of larvae, were used to construct a phylogeny. The monophyly of the subfamily was supported, and some generic‐level sister‐group relationships proposed in the literature, but without previous cladistic analyses, were also supported. The genus Blaesoxipha Loew, as currently recognized, was not monophyletic in our analysis. The genus Helicobia Coquillett is synonymized with Sarcophaga Meigen syn. nov. and treated as a subgenus of the latter. The Sarcophaga subgenera Neobellieria Blanchard and Mehria Enderlein were not monophyletic. Many of the clades in the analysis were supported primarily or exclusively by male genitalic character states, highlighting the importance of the male genitalia as a source of morphological characters for sarcophagine phylogeny. © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 158 , 740–778.     

Phylogeny of Heteronychia: the largest lineage of Sarcophaga (Diptera: Sarcophagidae)

Sarcophaga Meigen is one of the megadiverse genera of true flies, with approximately 850 valid species worldwide. The genus is divided into about 160 subgenera, the validity of a vast majority of which has never been verified using cladistic methods. This paper deals with the mainly Palaearctic subgenus Heteronychia Brauer & Bergenstamm, which comprises 89 species and is thus the largest subunit of Sarcophaga. We performed a cladistic analysis of the group based exclusively on male morphological characters. Parsimony analyses were run on a matrix of 84 characters for 88 species. Species of the subgenera Discachaeta Enderlein and Notoecus Stein were also included in the matrix. A further analysis was carried out using a subset of characters from the terminalia alone (70 characters). The results show that the clade formed by Heteronychia, Discachaeta, and Notoecus is monophyletic, with Discachaeta emerging as polyphyletic whereas Sarcophaga (Notoecus) longestylata Strobl is nested within the Sarcophaga filia‐group. Character states supporting Heteronychia and the few well‐supported species‐groups are discussed in detail. The following synonymies are proposed: Discachaeta = Heteronychia ( syn. nov. ) and Notoecus = Heteronychia ( syn. nov. ). The paper also includes a historical background of the taxon in relation to the classification of the genus Sarcophaga over the past two centuries, as well as a terminological review of the male terminalia, particularly of the distiphallus. © 2013 The Linnean Society of London     

Molecular phylogeny of the hyperdiverse genus Sarcophaga (Diptera: Sarcophagidae), and comparison between algorithms for identification of rogue taxa

The hyperdiverse genus Sarcophaga Meigen, with about 890 valid species arranged within 169 subgenera, accounts for almost half of the diversity of the subfamily Sarcophaginae. Current phylogenetic hypotheses for this genus are poorly supported or based on small taxon sets, or both. Here, we use molecular data from the genes COI and 28S to reconstruct the phylogeny of Sarcophaga based on the most comprehensive sampling for the group to date: 144 species from 47 subgenera, including representatives from all regional faunas for the first time. Of the total sequences of Sarcophaga used in the present study, 94.7% were newly generated. The secondary structure of the D1–D3 expansion segments of 28S is presented for the first time for the family Sarcophagidae, and is used in a multiple sequence alignment. Branch support and tree resolution increased remarkably through rogue taxa identification and exclusion. Rogue behaviour was explained mostly as a missing data problem. The RogueNaRok web service and the algorithms chkmoves, IterPCR and prunmajor implemented in the computer program TNT were equally good at identifying critical rogue species, but chkmoves and IterPCR also identified rogue clades. Pruning rogues increased the number of monophyletic subgenera in consensus trees from one to six out of 19 subgenera with more than one representative species. Bayesian inference, maximum‐likelihood and parsimony analyses recovered more monophyletic subgenera after the removal of rogue taxa, with parsimony showing the largest improvements in branch support and resolution. Although with low support, Nearctic taxa were found to be the earliest diverging lineages, followed by a subsequent diversification of Old World faunas, which is in agreement with currently available evidence of a New World origin and early diversification of Sarcophaga.     

Phylogenetic relationships of flesh flies in the subfamily Sarcophaginae based on three mtDNA fragments (Diptera: Sarcophagidae)

In an effort to improve our knowledge of the phylogenetic relationships among species and genera of the subfamily Sarcophaginae, we analysed data from three mitochondrial gene fragments. Sequence data for portions of the genes cytochrome oxidase I (COI), cytochrome oxidase II (COII) and dehydrogenase subunit 4 (ND4) were obtained from 43 species of Sarcophagidae representing 15 genera. We used a Bayesian approach to simultaneously choose how best to partition the data and which substitution model to apply to each partition. Phylogenetic relationships were inferred using Bayesian Inference and Maximum Likelihood methods. Our results are consistent with monophyly of the subfamily Sarcophaginae (posterior probability 1; bootstrap support 93%), as well as with monophyly of several genera within the Sarcophaginae (including Sarcophaga s.l.; posterior probability 1; bootstrap support 97%). We found support for a sister‐group relationship between Ravinia Robineau‐Desvoidy and Oxysarcodexia Townsend, which has been hypothesised by past authors on the basis of morphological similarities, although this was supported only in the Bayesian analyses (posterior probability 0. 81–0. 98), and for some novel supra‐generic clades. Contrary to a recent morphological hypothesis, we do not find Helicobia Coquillett to be nested within Sarcophaga Meigen; our data suggest, but do not strongly support, a hypothesis that Peckia Robineau‐Desvoidy is the sister group to Sarcophaga.     

Immunohistochemical localisation of the hypertrehalosaemic hormone II (Cam-HrTH-II) and related peptides in the nervous system of Carausius morosus and Sarcophaga bullata

Summary A polyclonal antiserum was prepared against an N-terminal modified Cam-HrTH-II (Leu-Asn-Phe-...), one of the members of the large AKH/RPCH peptide family, first isolated from Carausius morosus. The localisation of this peptide was performed by means of immunocytochemical methods in the brain and corpora cardiaca-corpora allata complex of the stick insect, Carausius morosus and the grey fleshfly, Sarcophaga bullata. The distribution patterns of molecules reactive to the Cam-HrTH-II and the LomAKH-I antisera in both insect species were compared. In Carausius, both antisera reacted in the same cell bodies. In Sarcophaga, some neurons were stained by both, others only by one of the two antisera. By combining two different antisera, we demonstrated that there are no Lom-AKH-I-like molecules present in Carausius and that there must occur at least three different AKH-like molecules in the brain of Sarcophaga. One is similar to Cam-HrTH-II, the second to Lom-AKH-I and the third is an AKH/RPCH-like peptide, different from Lom-AKH-I and Cam-HrTH-II.     

The effects of two parasitoids on the life history and metapopulation structure of the intertidal snail Littoraria filosa in different-sized patches of mangrove forest

We describe differences in life history of the intertidal arboreal snail, Littoraria filosa, among patches of mangroves ranging in size from isolated trees to large stands several square kilometres in area. Recruitment of L. filosa occurred from mid spring (October) to early winter (June), recruits grew rapidly and copulating adults were found during the following September–April. Populations within large patches of forest were annuals; all or most individuals died between October–January (spring–midsummer). In contrast, those in smaller peripheral patches were more likely to survive over the summer but survival differed among patches and years. These differences in life history were caused by a parasitoid fly (genus Sarcophaga) that attacked L. filosa 10 mm and longer and was present in all large patches, but absent from, or rare, in smaller peripheral patches. Experimental introductions to isolated trees confirmed that the fly could kill L. filosa. Another sarcophagid parasitoid that attacked L. filosa from 4 to less than 10 mm long was also found in every patch. The combined effects of these parasitoids appear to determine the metapopulation structure of L. filosa. Most adults in large patches were killed by the larger fly during early summer. Summer recruits were often killed by the smaller fly within a month of settlement and when this happened effective recruitment of L. filosa was reduced to autumn. The planktotrophic larval stage of L. filosa lasts less than 1 month, so the source of autumn recruits to all patches must have been adults that survived the early summer, most of which were in small patches or on isolated trees. Consequently these ”peripheral sources” are likely to be important for persistence of the metapopulation of L. filosa. The results of this study demonstrate that metapopulation structure may be determined by complex interactions and that common models cannot be assumed to apply in all habitats. Received: 15 September 1999 / Accepted: 31 January 2000     

Insect oostatic activity of GnRH and its fragments

Mammal, ¹²⁵I-mammal, salmon, chicken I and II GnRHs and three fragments of mammal GnRH were synthesized and their effect on oogenesis in the flesh fly Neobellieria(formerly Sarcophaga) bullata (Diptera) was investigated. The peptides were prepared by the Merrifield solid phase synthesis on polystyrene/divinylbenzene polymer using the N-Boc strategy in DMF and were purified by preparative RP-HPLC in a gradient of water-MeOH. From the peptides assayed, only mammal GnRH and two of its carboxy-terminus truncated analogs remarkably affected the processes of egg development in ovarioles, causing changes in the follicular epithelium, proliferation of its nuclei and cell division towards the inner part of the egg chamber. The process led to the occurrence of multinuclear follicular epithelium which finally filled up almost the whole egg chamber and then it degenerated. The inability of GnRH of other animal species to evoke the changes in the egg development establishes the question of primary structures of GnRH responsible for these biological effects. The identityof sequences of GnRHs from position 1 up to 6 (with the exception of chicken GnRH II) points to functionality of amino acids located in positions 7 and 8 of the peptide chain. The radioactivity of the ¹²⁵I-labelled mammal GnRH with maintained oostatic activity and its receptor competition with the non-labelled mammal GnRH were measured in selected insect organs and exhibited different residual values according to the organ and the time after applicationof the peptide. A transfer of the radioactivity into the next (F₁) generation was also observed.     

Influences of blackberry margins on population dynamics of Drosophila suzukii and grape infestation in adjacent vineyards

Understanding the dynamics of pest insect populations in relation to the presence of non‐crop habitats and infestation levels of adjacent crops is essential to develop sustainable pest management strategies. The invasive pest species Drosophila suzukii (Diptera: Drosophilidae) is able to utilize a broad range of host plants. In viticulture, scientific risk assessment for D. suzukii has only recently started and studies assessing the effects of field margins containing wild host plants on D. suzukii population dynamics and on infestation risks in adjacent vineyards are lacking. Thus, in a one‐year field study, the role of different field margins on fly abundance and crop infestation in adjacent vineyards of Vitis vinifera, variety “Pinot Noir,” were investigated. Different monitoring methods were conducted to assess fly distribution, sex ratio and grape infestation in 14 vineyards adjacent to field margins containing either blackberry (BB) Rubus spp. or non‐host (NH) plants. Our results show that blackberries strongly enhanced D. suzukii abundance within field margin vegetation all year long, whereas fly abundance in vineyards adjacent to BB margins was just enhanced in some seasonal periods. Moreover, the influence of BB margins was limited by distance. However, high fly numbers in BB field margins did result in zero egg infestation of “Pinot Noir” berries. These results may have important implications for winegrowers to make efficient management decisions: regardless of high abundance of adult D. suzukii, only grape berry monitoring can assess the actual infestation risk and the potential need to take management action.     

Population fluctuations and interspecific competition between Tephritid flies attacking fruit crops in the New Valley oases,Egypt

Population fluctuations of the Mediterranean fruit fly Ceratitis capitata (Wiedemann) and the peach fly Bactrocera zonata (Saunders) were monitored with lure trap collections in three provinces in the New Valley oases, Western Desert, Egypt. Results showed marked temporal differences in peak trap catches of the two flies in the selected sites all over the entire studied areas. One annual peak of C. capitata was recorded during both October 2005 and February 2006 and coincided with the ripening period of citrus trees in Kharga oases. However, two annual peaks were recorded during June and September 2005 in Bodkholow province and coincided with the ripening period of apple and mango. On the other hand, two annual peaks of B. zonata were recorded in Kharga oases throughout May and September and coincided with the ripening periods of apricot, mango and guava. One annual peak only was recorded in the round up of September and/or October in both Moot and Bodkholow in Dakhla oases and coincided with the ripening period of mango, guava and citrus. The occurrence of C. capitata was very limited in comparison with B. zonata. Population fluctuations of the two pests in the studied sites were significantly different. The ability of the traps used in capturing both C. capitata and B. zonata indicated that the yellow sticky trap was more effective in capturing C. capitata. However, the Abdel-Kawi trap was significantly efficient at trapping B. zonata. The occurrence of B. zontata in high numbers all over the study period compared to C. capitata is considered as good proof that this invading fly may be considered as a vigorous competitive tephritid fly to the native fly C. capitata.     

Evaluation of Beauveria bassiana applications against adult house fly, Musca domestica, in commercial caged-layer poultry facilities in New York state

Applications of a commercially produced Beauveria bassiana product, balEnce, were compared with pyrethrin treatments for the control of adult house flies in New York high-rise, caged-layer poultry facilities. An integrated fly management program, which included the release of house fly pupal hymenopteran parasitoids, was used at all facilities. Adult house fly populations were lower in B. bassiana-treated facilities during the spray and post-spray periods, as recorded on spot cards. Concurrently, the numbers of house fly larvae recovered in B. bassiana-treated facilities were less than one-half that of the pyrethrin-treated facilities. House fly pupal parasitism levels were low, but similar under both treatment regimes. The numbers of adult and larval Carcinops pumilio, a predatory beetle, recovered from B. bassiana-treated facilities were 43 and 66% greater than from the pyrethrin-treated facilities, respectively.     

The effects of larval competition on puparial size in Sarcophaga spp. (Diptera: Sarcophagidae) breeding in dead snails

An account is given of the effects of competition for food on the volume of the puparia of three species of Sarcophaga breeding in dead snails. The relationships between the number of puparia in a snail and the mean and total volume of the puparia, and the extent to which food is equally divided among the larvae are discussed.     

The circadian eclosion rhythm in Sarcophaga argyrostoma: A limit cycle representation of the pacemaker

We examine the effects of light upon the phase and coherence of the Sarcophaga eclosion rhythm and show that the limit cycle representation developed previously for the mosquito Culex pipiens quinquefasciatus can be extended, with very slight modification, to Sarcophaga.     

Biological control of <Emphasis Type="Italic">Musca domestica</Emphasis> and <Emphasis Type="Italic">Stomoxys calcitrans</Emphasis> by mass releases of the parasitoid <Emphasis Type="Italic">Spalangia cameroni</Emphasis> on two Norwegian pig farms

House flies (Musca domestica L.) and stable flies (Stomoxys calcitrans (L.)) (Diptera: Muscidae) are nuisance pests on livestock farms. In the present study we tested the effect of biweekly mass release of Spalangia cameroni Perkins (Hymenoptera: Pteromalidae), the most common parasitoid of house flies and stable flies in southern Norway, on pig premises with scattered fly breeding sites. The study spanned two successive summer periods, with two control and two release units each year. Spalangia cameroni suppressed both house flies and stable flies in one release unit during the first year, and stable flies in one release unit the second year. The apparent lack of success in fly control in the other release units was likely due to immigration of flies in two of the trials and high temperatures in one trial. Recommendations concerning release of S. cameroni in similar pig production premises are given. Handling editor: Torsten Meiners.     

Diel Calling Activity and Field Survival of the Bushcricket,Sciarasaga quadrata (Orthoptera: Tettigoniidae): A Role for Sound-locating Parasitic Flies?

In this study I examine the variability in calling activity of Sciarasaga quadrata over both the short (4 d) and long term (life span), and the temporal pattern to host searching by its most significant natural enemy, the ormiine fly Homotrixa alleni. Indeed, few male S. quadrata survive the entire calling season as this acoustically orienting parasitoid fly continually ‘culls’ calling males out of the population. I found that male S. quadrata commence calling over 1 h before sunset, cease around midnight and call, on average, for over 3 h within this period (range 0–11 h 09 min). There was significant concordance and repeatability in calling activity over both the short and long term, enabling me to conclude that relative differences in calling activity among males persist throughout a male's life span. There was a distinct peak to host searching by gravid female flies, with 82 % of all flies collected at acoustic traps between sunset and midnight. A positive association between call duration (time spent calling) and fly attraction was evident from multiple collections of flies at single traps. Counter to expectation, there was no significant decline in average call duration across successive collections of males, indicating that long-call-duration males were not being selectively culled by flies from the calling population. Males at the end of the calling season, when their remaining reproductive potential is low, allocate the majority of their calling time between sunset and midnight despite the high risk of fly attack. The distribution of call durations within the population was not skewed, which if heritable, indicates a lack of history of directional selection acting on call duration. Limited evidence suggests that encounters with females are rare in S. quadrata so that males may be selected, irrespective of natural selection pressure, to call for long periods of time to optimize their chance of attracting mates. The above findings are further discussed in relation to parasitoid foraging and acoustic reproductive strategies.     

Neuropeptidomics of the Australian sheep blowfly Lucilia cuprina (Wiedemann) and related Diptera

Insect neuropeptides are the most diverse and important group of messenger molecules that regulate almost all physiological processes, including behavior. In this study, we performed a combination of matrix assisted laser desorption ionization time of flight (MALDI-TOF) and electrospray ionization quadrupole time of flight (ESI-Q-TOF) mass spectrometry to analyze the peptidome of the brain and the neurohemal organs of the Australian sheep blowfly Lucilia cuprina and compared the data with those of related flies such as the gray flesh fly Sarcophaga (=Neobellieria) bullata; the cabbage root fly Delia radicum, the fruit fly Drosophila melanogaster, and the yellow fever mosquito, Aedes aegypti. Without counting low intensity signals of truncated peptides, 45 neuropeptides arising from 12 neuropeptide genes (adipokinetic hormone, CAPA-peptides, corazonin, extended FMRFamides, SIFamide, insect kinin, short neuropeptide F, NPLP-1 peptides, HUGIN-pyrokinin, sulfakinins, allatostatins A, putative eclosion hormone precursor peptide) were identified; sequences of extended FMRFamides were reported in a separate publication. The remarkable similarity of the peptidome of cyclorraphan flies, which contain a large number of ecologically important species, does not support the development of a species-specific neuropeptide-based insect pest control strategy. However, mass spectrometric approaches as shown here do not cover the entire peptidome or differences at the receptor level and it is possible that group-specific peptide ligands or receptors exist that escaped the detection.     

Tolerance and antibiosis resistance to cabbage root fly in vegetable Brassica species

Four accessions of the wild species Brassica fruticulosa Cyrillo (Brassicaceae) were studied in order to identify its tolerance and antibiosis resistance to the cabbage root fly, Delia radicum L. (Diptera: Anthomyiidae), in comparison to a widely cultivated cauliflower cultivar and a rapid cycling Brassica oleracea L. line. Antibiosis was prominent, as the insects reared on resistant accessions showed reduced individual pupal weight, total pupal weight, adult dry weight, and the longest average fly eclosion time. Host plant resistance, however, did not affect the sex ratio of adult flies. A study of the root architecture of plants with and without root fly inoculation revealed differences in the structure within B. oleracea accessions. A long main root and a high number of lateral roots appeared to be important characteristics for a Brassica type, with a higher tolerance level to cabbage root fly attack.     

Successional patterns of the insect fauna on a pig carcass in southern Italy and the role of Crematogaster scutellaris (Hymenoptera,Formicidae) as a carrion invader

The sarcosaprophagous fauna plays a key role in organic matter decomposition. Moreover, the biological, ecological and behavioral specificities of the taxa are useful to reconstruct the decay history of a corpse or carcass, often back to the lethal event. Here we report the seasonal succession of the insect fauna on a pig carcass exposed in a rural area in Calabria (southern Italy) during summer 2007 and 2008. The aim is to identify and qualitatively assess the major taxa of forensic importance in this region. The principal fly invaders were Lucilia caesar (L.), L. sericata (Meigen, 1826), Chrysomya albiceps (Wiedemann, 1819), Sarcophaga (Meigen, 1826) spp. and Amobia (Robineau‐Desvoidy, 1830) spp., Musca domestica (L.) and Muscina stabulans (Fallen, 1817). The primary beetle species collected in summer belonged to Dermestidae, Dermestes maculatus (De Geer, 1774) and Cleridae, Necrobia rufipes (De Geer, 1775). This paper also examined the ecological role of ants in the insect succession and describes the evidence of skin injuries directly inflicted by the acrobat ant Crematogaster (Acrocoelia) scutellaris (Olivier, 1791) (Hymenoptera Formicidae) while feeding on pig carrion. Ants belonging to two other species were also collected: Camponotus aethiops (Latreille, 1798) and Tetramorium semilaeve (André, 1881). Ants can invade carcasses and corpses directly, disrupting blowfly egg laying or preying on their larvae. Our data on the carrion faunal composition and role of ants as invaders should be useful for further forensic cases in Calabria (southern Italy). This is among the few reports of ants as forensically relevant species.