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BackgroundDiabetes and its concurrent complications impact a significant proportion of the population of the US and create a large financial burden on the American health care system. FDA-approved maggot debridement therapy (MDT), the application of sterile laboratory-reared Lucilia sericata (green bottle fly) larvae to wounds, is a cost-effective and successful treatment for diabetic foot ulcers and other medical conditions. Human platelet derived growth factor-BB (PDGF-BB) is a secreted dimeric peptide growth factor that binds the PDGF receptor. PDGF-BB stimulates cell proliferation and survival, promotes wound healing, and has been investigated as a possible topical treatment for non-healing wounds. Genetic engineering has allowed for expression and secretion of human growth factors and other proteins in transgenic insects. Here, we present a novel concept in MDT technology that combines the established benefits of MDT with the power of genetic engineering to promote healing. The focus of this study is to create and characterize strains of transgenic L. sericata that express and secrete PDGF-BB at detectable levels in adult hemolymph, whole larval lysate, and maggot excretions/ secretions (ES), with potential for clinical utility in wound healing.ResultsWe have engineered and confirmed transgene insertion in several strains of L. sericata that express human PDGF-BB. Using a heat-inducible promoter to control the pdgf-b gene, pdgf-b mRNA was detected via semi-quantitative PCR upon heat shock. PDGF-BB protein was also detectable in larval lysates and adult hemolymph but not larval ES. An alternative, tetracycline-repressible pdgf-b system mediated expression of pdgf-b mRNA when maggots were raised on diet that lacked tetracycline. Further, PDGF-BB protein was readily detected in whole larval lysate as well as larval ES.ConclusionsHere we show robust, inducible expression and production of human PDGF-BB protein from two conditional expression systems in transgenic L. sericata larvae. The tetracycline-repressible system appears to be the most promising as PDGF-BB protein was detectable in larval ES following induction. Our system could potentially be used to deliver a variety of growth factors and anti-microbial peptides to the wound environment with the aim of enhancing wound healing, thereby improving patient outcome in a cost-effective manner.

The parasitic mite Varroa destructor Anderson & Trueman (Acari: Varroidae) has plagued European honey bees, Apis mellifera L. (Hymenoptera: Apidae), in the Americas since its introduction in the 1980s. For many years, these mites were sufficiently controlled using synthetic acaricides. Recently, however, beekeepers have experienced increased resistance by mites to chemical pesticides, which are also known to leave residues in hive products such as wax and honey. Thus there has been increased emphasis on nonchemical integrated pest management control tactics for Varroa. Because mites preferentially reproduce in drone brood (pupal males), we developed a treatment strategy focusing on salvaging parasitized drones while removing mites from them. We removed drone brood from colonies in which there was no acaricidal application and banked them in separate "drone-brood receiving" colonies treated with pesticides to kill mites emerging with drones. We tested 20 colonies divided into three groups: 1) negative control (no mite treatment), 2) positive control (treatment with acaricides), and 3) drone-brood removal and placement into drone-brood receiving colonies. We found that drone-brood trapping significantly lowered mite numbers during the early months of the season, eliminating the need for additional control measures in the spring. However, mite levels in the drone-brood removal group increased later in the summer, suggesting that this benefit does not persist throughout the entire season. Our results suggest that this method of drone-brood trapping can be used as an element of an integrated control strategy to control varroa mites, eliminating a large portion of the Varroa population with limited chemical treatments while retaining the benefits of maintaining adult drones in the population.

The pine bark adelgid, Pineus strobi Hartig (Hemiptera: Adelgidae), is a native herbivore of eastern white pine, Pinus strobus L. (Pinales: Pinaceae), in eastern North America. P. strobi does not appear to have any dominant overwintering lifestage in southwest Virginia, as it does in its northern range. Eggs can be found consistently from late March through early December and may be produced sporadically later throughout the winter during warm periods. Two distinct generations were observed in the spring, after which life stage frequencies overlapped. Adult body size varied seasonally and was greatest in the spring. The present study constitutes the first recording of phenological details of the P. strobi in its southern range, informing biological control efforts aimed at closely related invasive pests. The phenological plasticity observed between northern and southern P. strobi populations provides insight into the potential effects of climate on the population dymanics of this and related species.

The pine bark adelgid, Pineus strobi (Hartig) (Hemiptera: Adelgidae), is an herbivore native to eastern North America that specialises on eastern white pine, Pinus strobus Linnaeus (Pinaceae). Little is known about P. strobi, especially in its southern range in the Appalachian Mountains, United States of America, and the composition of its predator complex has not yet been documented in this region. The current study identifies arthropod predators associated with P. strobi in Appalachian forests of Virginia based on a two-year survey. Predators were identified using morphology and DNA barcoding. Predator species include: Laricobius rubidus LeConte (Coleoptera: Derodontidae), Leucopis piniperda Malloch (Diptera: Chamaemyiidae), and Leucopis argenticollis Zetterstedt (Diptera: Chamaemyiidae), that are known adelgid specialists. Also found were predators from the families Cecidomyiidae (Diptera), Coccinellidae (Coleoptera), Chrysopidae (Neuroptera), Hemerobiidae (Neuroptera), and Syrphidae (Diptera). The Cecidomyiidae were especially diverse, with 14 different species inferred from their DNA barcodes. Knowledge of this predator complex is particularly valuable for anticipation and detection of potential interactions between native predator species and those that are being considered for the introduction for biological control of invasive adelgid pests within the southern Appalachian ecosystem.