Lucilia Sericata Research Articles

The composition of the cuticular and internal free fatty acids and alcohols from Lucilia sericata males and females.

GC, GC–MS, and HPLC–LLSD analyses were used to identify and quantify cuticular and internal lipids in males and females of the blow-fly (Lucilia sericata). Sixteen free fatty acids, seven alcohols and cholesterol were identified and quantitatively determined in the cuticular lipids of L. sericata. Cuticular fatty acids ranged from C6 to C20 and included unsaturated entities such as 16:1n-9, 18:1n-9, 20:4n-3 and 20:5n-3. Cuticular alcohols (only saturated and even-numbered) ranged from C12 to C20 in males and C10 to C22 in females. Only one sterol was found in the cuticular lipids of both males and females. 23 free fatty acids, five alcohols and cholesterol were identified in the internal lipids. Internal fatty acids were present in large amounts—7.4 mg/g (female) and 10.1 mg/g (male). Only traces of internal alcohols (from C14 to C26 in males, from C14 to C22 in females) were found in L. sericata. Large amounts of internal cholesterol were identified in L. sericata males and females (0.49 and 0.97 mg/g of the insect body, respectively).

A Case of Traumatic Myiasis in a Domestic Rabbit (Oryctolagus cuniculus) Caused By Lucilia sericata

Lucilia sericata is one of the factors resulting in facultative traumatic myiasis in animals and humans. L. sericata threatens human health and leads to significant economic losses in animal industry by leading to serious parasitic infestations. A three month old female rabbit was presented to the clinics of the Veterinary Faculty of Dicle University for the treatment of the wound located on the left carpal joint. The examination revealed that the wound was infested with larvae. The microscopic inspection of the larvae collected from the rabbit showed that they were the third instar larvae of L. sericata.

Expression of a cGMP compatibleLucilia sericatainsect serine proteinase debridement enzyme

Previously, we demonstrated the effectiveness of a research grade recombinant chymotrypsin, derived from the larvae of Lucilia sericata, in "debriding" slough/eschar from venous leg ulcers ex vivo. Furthermore, we were able to formulate this enzyme for successful delivery to in vitro wound healing assays, from a prototype hydrogel wound dressing, and showed that enzyme delivered in this way could degrade wound tissue ex vivo. Recently, to progress biotechnological development of the enzyme as a potential therapeutic product, we explored expression using current good manufacturing practice (cGMP) guidelines, and now report that a recombinant chymotrypsin I zymogen from L. sericata can be expressed in the cGMP acceptable strain of Escherichia coli (BLR-DE3). In addition, the conditions required for purification, refolding and activation of the chymotrypsinogen have been determined. The activated enzyme was stable, and effective in digesting wound slough/eschar tissue. To summarise, we have successfully initiated the production and characterisation of a novel cGMP compatible product for use in future clinical trials.

Disentangling the functional consequences of the connectivity between optic-flow processing neurons

Typically, neurons in sensory areas are highly interconnected. Coupling two neurons can synchronize their activity and affect a variety of single-cell properties, such as their stimulus tuning, firing rate or gain. All of these factors must be considered to understand how two neurons should be coupled to optimally process stimuli. We quantified the functional effect of an interaction between two optic-flow processing neurons (Vi and H1) in the fly (Lucilia sericata). Using a generative model, we estimated a uni-directional coupling from H1 to Vi. Especially at a low signal-to-noise ratio (SNR), the coupling strongly improved the information about optic-flow in Vi. We identified two constraints confining the strength of the interaction. First, for weak couplings, Vi benefited from inputs by H1 without a concomitant shift of its stimulus tuning. Second, at both low and high SNR, the coupling strength lay in a range in which the information carried by single spikes is optimal.

Plant Terpenoids: Acute Toxicities and Effects on Flight Motor Activity and Wing Beat Frequency in the Blow Fly Phaenicia sericata

We evaluated the acute toxicities and the physiological effects of plant monoterpenoids (eugenol, pulegone, citronellal and alpha-terpineol) and neuroactive insecticides (malathion, dieldrin and RH3421) on flight muscle impulses (FMI) and wing beat signals (WBS) of the blow fly (Phaenicia sericata). Topically-applied eugenol, pulegone, citronellal, and alpha-terpineol produced neurotoxic symptoms, but were less toxic than malathion, dieldrin, or RH3421. Topical application of eugenol, pulegone, and citronellal reduced spike amplitude in one of the two banks of blow fly dorsolongitudinal flight muscles within 6-8 min, but with citronellal, the amplitude of FMIs reverted to a normal pattern within 1 hr. In contrast to pulegone and citronellal, where impulse frequency remained relatively constant, eugenol caused a gradual increase, then a decline in the frequency of spikes in each muscle bank. Wing beating was blocked permanently within 6-7 min of administering pulegone or citronellal and within 16 mins with eugenol. alpha-Terpineol-treated blow flies could not beat their wings despite normal FMI patterns. The actions of these monoterpenoids on blow fly flight motor patterns are discussed and compared with those of dieldrin, malathion, RH3421, and a variety of other neuroactive substances we have previously investigated in this system. Eugenol, pulegone and citronellal readily penetrate blow fly cuticle and interfere with flight muscle and/or central nervous function. Although there were differences in the effects of these compounds, they mainly depressed flight-associated responses, and acted similarly to compounds that block sodium channels and facilitate GABA action.

The use of maggot debridement therapy in the treatment of chronic wounds in hospitalised and ambulatory patients

To summarise our experience of the use of medicinal maggots for the debridement of necrotic chronic wounds and to try and identify prognostic factors for debridement success and associated pain. During the years 1996–2009, 723 wounds of 435 patients (180 females and 255 males) were treated with maggot debridement therapy (MDT) in 16 departments and units of the Hadassah Hospital in Jerusalem, Israel. Overall, 261 patients were treated during hospitalisation, while 174 were treated as ambulatory patients. In 90.5% of the patients the wounds were located on the leg, but only 48.0% had diabetic foot ulcers. The wound duration range from one to 240 months (mean=8.9; median=4 months). Sterile maggots of the green bottle fly, Lucilia sericata, were used for MDT. In 90.6% of the cases, maggots were placed directly on the wound using a cage-like dressing and left for 24 hours, while in 9.4% of the patients maggots concealed in a tea-bag like polyvinyl netting were used. The concealed maggots were left on the wound for 2–3 days. The number of treatments was 1–48 (mean=2.98; median=2) and the duration of the treatment varied between one and 81 days (mean=4.65; median=3). In 357 patients (82.1%) complete debridement of the wound was achieved, while in 73 patients (16.8%) the debridement was partial and in five (1.1%) it was ineffective. Increased pain or discomfort during MDT were reported in 38% of the patients. MDT is a very safe, simple and effective treatment modality for chronic wounds in ambulatory and hospitalised patients. There were no external sources of funding for this study. The authors have no additional conflicts of interest to declare.

Ade novotranscriptome assembly ofLucilia sericata(Diptera: Calliphoridae) with predicted alternative splices, single nucleotide polymorphisms and transcript expression estimates

The blow fly Lucilia sericata (Diptera: Calliphoridae) (Meigen) is a nonmodel organism with no reference genome that is associated with numerous areas of research spanning the ecological, evolutionary, medical, veterinary and forensic sciences. To facilitate scientific discovery in this species, the transcriptome was assembled from more than six billion bases of Illumina and twenty-one million bases of 454 sequence derived from embryonic, larval, pupal, adult and larval salivary gland libraries. The assembly was carried out in a manner that enabled identification of putative single nucleotide polymorphisms (SNPs) and alternative splices, and that provided expression estimates for various life history stages and for salivary tissue. The assembled transcriptome was also used to identify transcribed transposable elements in L. sericata. The results of this study will enable blow fly biologists, dipterists and comparative genomicists to more rapidly develop and test molecular and genetic hypotheses, especially those regarding blow fly development and salivary gland biology.

Cuticular and internal n-alkane composition ofLucilia sericatalarvae, pupae, male and female imagines: application of HPLC-LLSD and GC/MS-SIM

The composition of cuticular and internal n-alkanes in Lucilia sericata larvae, pupae, and male and female imagines were studied. The cuticular and internal lipid extracts were separated by HPLC-LLSD, after which the hydrocarbon fraction was identified by GC/MS in selected ion monitoring (SIM) and total ion current (TIC) modes. The cuticular lipids of the larvae contained seven n-alkanes from C23 to C31. The major n-alkane in L. sericata larvae was C29 (42.1%). The total cuticular n-alkane content in the cuticular lipids was 31.46 μg g-1 of the insect body. The internal lipids of L. sericata larvae contained five n-alkanes ranged from C25 to C31. The most abundant compound was C27 (61.71 μg g-1 of the insect body). Eighteen n-alkanes from C14 to C31 were identified in the cuticular lipids of the pupae. The most abundant n-alkanes ranged from C25 to C31; those with odd-numbered carbon chains were particularly abundant, the major one being C29:0 (59.5%). Traces of eight cuticular n-alkanes were present. The internal lipids of L. sericata pupae contained five n-alkanes, ranging from C25 to C31. The cuticular lipids of female imagines contained 17 n-alkanes from C12 to C30. Among the cuticular n-alkanes of females, C27 (47.5%) was the most abundant compound. Four n-alkanes, with only odd-numbered carbon chains, were identified in the internal lipids of females. The lipids from both sexes of L. sericata had similar n-alkane profiles. The cuticular lipids of adult males contained 16 n-alkanes ranging from C13 to C31. C27 (47.9%) was the most abundant cuticular n-alkanes in males. The same n-alkanes only with odd-numbered carbon chains and in smaller quantities of C27 (0.1%) were also identified in the internal lipids of males. The highest amounts of total cuticular n-alkanes were detected in males and females of L. sericata (330.4 and 158.93 μg g-1 of the insect body, respectively). The quantities of total cuticular alcohols in larvae and pupae were smaller (31.46 μg g-1 and 42.08 μg g-1, respectively). The internal n-alkane contents of larvae, pupae, and male and female imagines were significantly higher than the cuticular n-alkane contents (153.53, 99.60, 360.06 and 838.76 μg g-1 of the insect body, respectively).

The efficacy of maggot debridement therapy – a review of comparative clinical trials

Over the last decade, maggot debridement therapy (MDT) has been recognized by many clinicians as a potential adjunct to conventional therapy, and many patients with non healing, chronic ulcers have been treated. Numerous case reports and case series have described the successful use of MDT in a variety of ulcers. However, comparative clinical trials and in particular randomized controlled trials investigating the efficacy of MDT are sparse. A systematic search in the literature showed three randomized clinical trials and five non randomized studies evaluating the efficacy of sterile Lucilia sericata applied on ulcers with various aetiologies. Of these, seven studies had debridement and/or healing as an outcome variable. When evaluating maggots as debriding agents, the studies report MDT as being significantly more effective than hydrogel or a mixture of conventional therapy modalities, including hydrocolloid, hydrogel and saline moistened gauze. However, the design of the studies was suboptimal, with important differences in the use of other therapies, such as compression, that may influence both debridement and healing between the compared groups, as well as inappropriately short follow-up times. The quality of the studies therefore makes it difficult to conclude that MDT shortens healing time. The poor quality of the data used for evaluating the efficacy of MDT highlights the need for more and better designed investigations.

Proteus mirabilis interkingdom swarming signals attract blow flies

Flies transport specific bacteria with their larvae that provide a wider range of nutrients for those bacteria. Our hypothesis was that this symbiotic interaction may depend on interkingdom signaling. We obtained Proteus mirabilis from the salivary glands of the blow fly Lucilia sericata; this strain swarmed significantly and produced a strong odor that attracts blow flies. To identify the putative interkingdom signals for the bacterium and flies, we reasoned that as swarming is used by this bacterium to cover the food resource and requires bacterial signaling, the same bacterial signals used for swarming may be used to communicate with blow flies. Using transposon mutagenesis, we identified six novel genes for swarming (ureR, fis, hybG, zapB, fadE and PROSTU_03490), then, confirming our hypothesis, we discovered that fly attractants, lactic acid, phenol, NaOH, KOH and ammonia, restore swarming for cells with the swarming mutations. Hence, compounds produced by the bacterium that attract flies also are utilized for swarming. In addition, bacteria with the swarming mutation rfaL attracted fewer blow flies and reduced the number of eggs laid by the flies. Therefore, we have identified several interkingdom signals between P. mirabilis and blow flies.

Trakeotomi Yarasında Lucilia Sericata’nın Neden Olduğu Miyazis: Türkiye’den Nadir Görülen Bir Olgu Sunumu

Although myiasis is not a frequently encountered case in otorhinolaryngology practice, reported cases often involved the nose, ear and pharynx in rank order. Myiasis of tracheotomy wound is extremely rare. Lucilia larvae tend to infest the wound tissue (e.g. tracheotomy) rather than healthy tissues. Older and lonely patients with tracheotomy should be controlled by primary health care periodically. We report a Lucilia sericata myiasis in tracheotomy wound of an 80-year-old male patient who lived alone and had decreased hearing and vision due to senility.

Determination of the limits of usability of auricle morphology for personnel identification

Using CO II sequences to identify common species of carrion-breeding flies and larvae. flies and larvae were collected on the corpses of rats in Zhengzhou district, DNA was extracted, CO II sequences were amplified and sequenced. Clustalx and MEGA 4.0 software were used to analyze the gene sequences and to construct the phylogenetic trees. There was no significant gene difference between adults and larvae. COII gene sequences could be used to identify Boettcherisca peregrina, Aldrichina grahami and Lucilia illustris but they could not distinguish Lucilia cuprina from the Lucilia sericata because of their close evolutionary distance and single nucleotide polymorphisms in aldrichina grahami and Lucilia illustris populations were found. CO II sequence of mtDNA in Zhengzhou district can be used effectively to identify some common species of carrion-breeding fly. The method is simple and accurate.

Common Green Bottle Fly or Sheep Blow Fly Lucilia sericata (Meigen) (Insecta: Diptera: Calliphoridae)

The common green bottle fly is a common visitor to carrion, feces, and garbage. Like many of the other blow flies, it plays an important role in forensic, medical, and veterinary science. Larvae or maggots help forensic scientists to determine the period of insect colonization as it relates to the time of death, aiding law enforcement in their investigations. Medical treatment using maggot therapy can help to heal infections that are otherwise incurable. Learn more in this 4-page fact sheet written by Matthew Anderson and Phillip E. Kaufman and published by the UF Department of Entomology and Nematology, October 2011. Reviewed September 2014. EENY-502/IN903: Common Green Bottle Fly or Sheep Blow Fly Lucilia sericata (Meigen) (Insecta: Diptera: Calliphoridae) (ufl.edu)

The use of COI barcodes for molecular identification of forensically important fly species in Germany

Deoxyribonucleic acid (DNA)-based insect identification has become a routine and accurate tool in forensic entomology. In the present study, we demonstrate the utility of the mitochondrial DNA cytochrome oxidase I gene "barcoding region" as a universal marker for molecular identification of forensically important Diptera. We analyzed 111 specimens belonging to 13 species originating from Frankfurt am Main, Germany (Calliphoridae: Calliphora vicina, Calliphora vomitoria, Lucilia ampullacea, Lucilia caesar, Lucilia illustris, Lucilia sericata, Lucilia silvarum, Phormia regina, Protophormia terraenovae; Piophilidae: Parapiophila vulgaris; Muscidae: Hydrotaea dentipes, Hydrotaea ignava, Hydrotaea similis). Intraspecific variation ranged from 0 to 1.17% and interspecific variation occurred between 1.17% and 15.21%. Although differences within species were generally less than among species, divergence percentages overlapped due to low interspecific nucleotide divergence of the recently separated sister species L. caesar and L. illustris. However, all species formed distinct monophyletic clades and thus the cytochrome oxidase 1 (COI) barcode has been shown suitable for clear differentiation and identification of forensically relevant Diptera in Germany.

Cuticular muscle attachment sites as a tool for species determination in blowfly larvae

First results of a new method for species determination in third instar larvae of saprophagous blowflies are introduced. Cuticular attachment sites of a limited number of transversal muscles are visualized for light microscopic analysis. After removing the muscles and staining the cuticle, the attachment sites become visible as laterally symmetrical segmental clusters of dark dots. The combined patterns of five such clusters, located in the second, third and fourth segments, show sufficient differences to allow reliable separation of externally very similar larval Lucilia sericata and Lucilia illustris as well as Calliphora vomitoria and Calliphora vicina, the most common saprophagous blowfly species in Europe. Species determination even in poorly conserved, discoloured and fragmented blowfly larvae becomes possible with this new method. The method can primarily be applied for postmortem interval (PMI) calculations in forensic entomology. Interspecific morphological similarity of the larvae and differences in growth rate make species determination an essential requisite for an exact PMI calculation.

A review of comparative aspects of myiasis in goats and sheep in Europe

Myiasis is the infestation of living vertebrate animals and humans by the larvae of certain fly species (Diptera), which feed on the host's living or dead tissues and body fluids. Myiasis in sheep and goats is a major animal welfare issue causing considerable pain and suffering and, if untreated, can result in serious tissue injury, productivity losses, reproduction losses and death. A number of oestrid flies cause myiasis of sheep and goats. These include the goat warble flies (Przhevalskiana silenus), and the sheep nasal bot flies (Oestrus ovis). P. silenus has been reported mainly from goats. In contrast, infection rates and larval burdens with O. ovis are generally higher in sheep than goats, following both natural and artificial infestations, despite the fact that both ruminant species can act as hosts. Traumatic or wound myiasis is caused mainly by calliphorid and sarcophagid flies. Most of the calliphorid species involved, such as Lucilia sericata and Chrysomya albiceps, are facultative parasites. The single obligate parasite causing traumatic myiasis of goats and sheep in Europe is the sarcophagid, Wohlfahrtia magnifica, Wohlfahrt's wound myiasis fly. All calliphorids and sarcophagids can infest sheep and goats and there is no known predisposition of certain genetic types for different hosts. Myiasis causing parasites are found worldwide and are especially widespread in the countries of Europe and Africa bordering the Mediterranean Basin. Because of the milk oriented husbandry systems applied in those areas, prevention and control of myiasis can be problematic, especially of lactating females in sheep and goat herds, due to the lengthy withdrawal periods for many of the insecticidal products. Like many other pest species, global warming could enable them to move northwards in Europe, with damaging consequences for the naïve hosts that would thereby be exposed.

Forensic entomology: Molecular identification of blowfly species (Diptera: Calliphoridae) in Portugal

Insects, in particular Calliphoridae species have a very important role in decomposition process and are the earliest insects to infest a corpse. An accurate morphological identification is essential but very difficult or even sometimes impossible to do. So, molecular identification provides a rapid and reliable method that can be done in all development stages. The potential of mitochondrial cytochrome oxidase subunit I (COI) is very well established. But in some species this gene is not effective. In this work, we used the ribosomal internal transcribed spacer 2 (ITS2) to complement COI data, demonstrating ITS2 effectiveness in insects’ identification. Blowflies of the family Calliphoridae ( Calliphora vicina, Calliphora vomitoria, Lucilia caesar and Lucilia sericata) were collected in Portugal. COI fragments permitted correct specimens identification using BLAST search for all blowflies, except for L. caesar because of the high similarity with Lucilia illustris. For these species, we used ITS2 sequences for species determination. This genetic marker analysis facilitated the differentiation of these two species. Our results indicate that it would be of great importance to increase the sequences collection to prevent incorrect identification and reinforce results validity.

EFFECTS OF THALLIUM ON THE LARVAL DEVELOPMENT OF LUCILIA SERICATA MEIGEN 1826 AND PMI ESTIMATION

Determination of larval growth rate of and forensic analysis of the age of Calliphoridae larvae on a corpse are useful evidence in legal investigations for the estimation of exact death time and time duration after death; post mortem interval. However many factors, such as temperature, tissue type and contamination of drugs and toxins, effect larval development of blow fly larvae and consequently the estimation of post mortem interval. The present study examined the larval growth rate of a forensically important blow fly species, Lucilia sericata Meigen 1826 in different concentrations (0,12; 0,25; 0,50; 1 and 2 μg/g) of toxic heavy metal Thallium under controlled laboratory conditions. Body length and weight, death ratio of larvae and pupa between experimental and control groups were compared. Results demonstrated that the development rate of larvae between uncontaminated and contaminated diets varies significantly. In short, they molted later, reached maximum length more slowly and sometimes produced significantly smaller pupae in contaminated food source. These results emphasized that the importance of determining the contamination rate of toxins in tissue for the forensic entomologist, while using development rates from standard curves based on larvae fed non-contaminated mediums.

XEvaluation of Anti-adherent Activity of Excretions of Irradiated Lucilia sericata Maggot and Certain Essential Oils against Some Pathogenic Bacterial Strains

ESSENTIAL OILS are widely used for their medicinal properties. They block adhesion and colonization of pathogenic microbes to epithelial cells which associated with bacterial resistance to antibiotics. So, this study investigates the effect of

Seasonal and habitat abundance and distribution of some forensically important blow flies (Diptera: Calliphoridae) in Central California

Seasonal and habitat calliphorid abundance and distribution were examined weekly for two years (2001-2003) in Santa Clara County, California, using sentinel traps baited with bovine liver. Of the 34,389 flies examined in three defined habitats (rural, urban, and riparian), 38% of the total catch represented Compsomyiops callipes (Bigot) and 23% represented Phormia regina (Meigen). Other flies collected in this survey included Calliphora vomitoria (Linnaeus), Calliphora latifrons (Hough), Lucilia sericata (Meigen), Lucilia cuprina (Wiedemann), and Lucilia mexicana (Macquart), which is a new record for the area. Multivariate MANOVA and ANOVA (P ≤ 0.05) analysis indicate significant seasonal habitat preference for all fly species examined. This information may be used to identify potentially forensically impo rtant fly species within Santa Clara County, California.