A Case of Oral Myiasis Caused by Lucilia sericata (Diptera: Calliphoridae) in Korea

We report here a case of oral myiasis in the Republic of Korea. The patient was a 37-year-old man with a 30-year history of Becker''s muscular dystrophy. He was intubated due to dyspnea 8 days prior to admission to an intensive care unit (ICU). A few hours after the ICU admission, 43 fly larvae were found during suction of the oral cavity. All maggots were identified as the third instars of Lucilia sericata (Diptera: Calliphoridae) by morphology. We discussed on the characteristics of myiasis acquired in Korea, including the infection risk and predisposing factors.     

Canine Wound Myiasis Caused by Lucilia sericata (Diptera: Calliphoridae) in Korea

Myiasis is a relatively common infection of animals kept as pets, although only 1 case of canine myiasis has been described so far in the Republic of Korea. In the present study, we report an additional case of canine wound myiasis with identification of its causative agent, Lucilia sericata. An 8-year-old male Siberian husky dog was referred with anorexia, vomiting, and diarrhea to the Chungbuk National University Veterinary Medical Center, Cheongju-si (city), Chungcheongbuk-do (province), Korea in July 2013. Physical examination indicated the patient had a deep wound filled with a maggot swarm as a left gluteal lesion. A total of 216 maggots were removed by forceps, and the wounded area was sponged with gauzes and disinfected with 70% alcohol and a povidone-iodine solution. After daily care and suturing the wound, the patient was discharged at day 19 after admission. Recovered worms possessed morphological characteristics similar to those of L. sericata, namely, a sub-cylindrical body with 6–8 lobed anterior spiracles, round shaped with a button surrounded by a peritremal ring with no gaps, and similar distances between dorsal, median, and outer papillae of the 12th segment. Additionally, cox1 partial sequences (528 bp) obtained in the present study showed 100% identity with those of L. sericata (GenBank no. {"type":"entrez-nucleotide","attrs":{"text":"KT272854.1","term_id":"992320795","term_text":"KT272854.1"}}KT272854.1). L. sericata is indicated as a pathogen of myiasis infection not only in humans, but also in animals kept as pets in Korea.     

Wound myiasis in a wild boar by Lucilia caesar (Diptera: Calliphoridae): First case and current status of animal myiasis by this species

The first case of myiasis caused by Lucilia caesar (L.) (Diptera: Calliphoridae) in a wild boar, Sus scrofa L. (Artiodactyla: Suidae) is described. The myiasis occurred in October 2019 in Southern Italy and the identification of the agent was based on adult male morphology. The wild boar had a wound on its right side, near the neck, which was largely infested by larvae. The ecology, distribution and current literature status about cases of animal myiasis by this species is also included.     

First Report of Myiasis Caused by Cochliomyia hominivorax (Diptera: Calliphoridae) in a Diabetic Foot Ulcer Patient in Argentina

Myiasis is usually caused by flies of the Calliphoridae family, and Cochliomyia hominivorax is the etiological agent most frequently found in myiasis. The first case of myiasis in a diabetic foot of a 54-year-old male patient in Argentina is reported. The patient attended the hospital of the capital city of Tucumán Province for a consultation concerning an ulcer in his right foot, where the larval specimens were found. The identification of the immature larvae was based on their morphological characters, such as the cylindrical, segmented, white yellow-coloured body and tracheas with strong pigmentation. The larvae were removed, and the patient was treated with antibiotics. The larvae were reared until the adults were obtained. The adults were identified by the setose basal vein in the upper surface of the wing, denuded lower surface of the wing, short and reduced palps, and parafrontalia with black hairs outside the front row of setae. The main factor that favoured the development of myiasis is due to diabetes, which caused a loss of sensibility in the limb that resulted in late consultation. Moreover, the poor personal hygiene attracted the flies, and the foul-smelling discharge from the wound favoured the female''s oviposition. There is a need to implement a program for prevention of myiasis, in which the population is made aware not only of the importance of good personal hygiene and home sanitation but also of the degree of implication of flies in the occurrence and development of this disease.     

Modelling the impact of climate change on spatial patterns of disease risk: sheep blowfly strike by Lucilia sericata in Great Britain

Understanding the spatial scale and temporal pattern of disease incidence is a fundamental prerequisite for the development of appropriate management and intervention strategies. It is particularly critical, given the need to understand the elevated risks linked to climate change, to allow the most likely changes in the distribution of parasites and disease vectors to be predicted under a range of climate change scenarios. Using statistical models, the spatial distribution and climatic correlates of a range of parasites and diseases have been mapped previously, but their development into dynamic, predictive tools is less common. The aim of the work described here, was to use a species distribution model to characterise the environmental determinants of the monthly occurrence of ovine cutaneous myiasis (blowfly strike) by Lucilia sericata, the most frequent primary agent of northern European myiasis, and to then use this model to describe the potential spatial changes that might be expected in response to predicted climate change in Great Britain. The model predicts that the range of elevated temperatures predicted by current climate change scenarios will result in an increase in the risk of strike and an elongated blowfly season. However, even for the most rapid warming scenario predictions over the next 70 years, strike is not predicted to occur throughout the winter. Nevertheless, in this latter case, parts of central and southern England are likely to become too hot and dry for strike by L. sericata, to persist in mid-summer. Under these conditions, it is possible that other, more pathogenic Mediterranean agents of myiasis, such as Wolfhartia magnifica, could potentially replace L. sericata. Where the phenology of strike is altered by climate change, as predicted here, significant changes to the timing and frequency of parasite treatments and husbandry practices, such as shearing, will be required to manage the problem. The results suggest that the modelling approach adopted here could be usefully applied to a range of disease systems.     

Efficient germ-line transformation of the economically important pest species Lucilia cuprina and Lucilia sericata (Diptera, Calliphoridae)

The green blowfly species Lucilia cuprina and Lucilia sericata are economically important pests for the sheep industries of Australia and New Zealand. L. cuprina has long been considered a good target for a genetic pest management program. In addition, L. sericata maggots are used in the cleaning of wounds and necrotic tissue of patients suffering from ulcers that are difficult to treat by other methods. Development of efficient transgenesis methods would greatly facilitate the development of strains ideal for genetic control programs or could potentially improve “maggot therapy”. We have previously reported the germ-line transformation of L. cuprina and the design of a “female killing system” that could potentially be applied to this species. However, the efficiency of transformation obtained was low and transformed lines were difficult to detect due to the low expression of the EGFP marker used. Here we describe an efficient and reliable method for germ-line transformation of L. cuprina using new piggyBac vector and helper plasmids containing the strong promoter from the L. cuprina hsp83 gene to drive expression of the transposase and fluorescent protein marker gene. We also report, for the first time, the germ-line transformation of L. sericata using the new piggyBac vector/helper combination.     

Cloacal myiasis by Lucilia spp. (Diptera: Calliphoridae) in a rooster (Gallus gallus domesticus) and two Harris's hawks (Parabuteo unicinctus)

In this study, cloacal myiasis caused by dipterans of Lucilia genus was found in a rooster (Gallus gallus domesticus) and two Harris's hawks (Parabuteo unicinctus) from Peru. Larval dipteran were collected and preserved in ethanol. Morphological analysis indicated two species: Lucilia sericata in the rooster and in one Harris's hawk, and Lucilia cuprina in the other Harris's hawk. Molecular analysis confirmed the diagnosis by amplification of the nucleotide sequences of the cytochrome c oxidase subunit 1 gene and internal transcribed spacer 2 region. The sequences were compared with sequence references from a public sequence database, which showed a 100% matched identity. This study demonstrated for first time cloacal myiasis by L. sericata in a domestic bird from Peru and in Harris's hawk. Also, for the first time, L. cuprina was found in a bird of prey.     

Comparison of traps for the control of sheep blowfly in the U.K.

The ability of three commercially available trap types to catch Lucilia (Diptera: Calliphoridae) blowflies was assessed on three sheep farms in southwest England in 2008. The aim was to evaluate their relative value for the control of ovine cutaneous myiasis (sheep blowfly strike) on farms. There was a highly significant difference between the total number of female Lucilia caught per day by the traps, with an Agrilure Trap (Agrimin Ltd, Brigg, U.K.) catching more than the other trap types (Rescue Disposable Fly Trap, Sterling International, Spokane, U.S.A.; Redtop Trap, Miller Methods, Johannesburg, South Africa). However, there was no significant difference between the traps in the numbers of female Lucilia sericata (Meigen) caught. Nevertheless, consideration of the rate at which female L. sericata were caught over time showed that the Agrilure trap did not begin catching until about 30 days after its initial deployment. It subsequently caught L. sericata at a faster rate than the other two traps. The data suggest that the freeze‐dried liver bait used in the Agrilure trap required a period of about 30 days to become fully rehydrated and decompose to the degree required to attract and catch L. sericata. Once the bait was attractive, however, the trap outperformed the other two traps in terms of the rate of L. sericata capture. The Agrilure trap would appear to be the most effective of the designs tested for use against sheep blowfly and blowfly strike in the U.K., but care would be needed to ensure that the traps were deployed in advance of the blowfly season so that the bait was suitably aged when trapping was required.     

Ophthalmomyiasis caused by a Phormia sp. (Diptera: Calliphoridae) larva in an enucleated patient

Ophthalmomyiasis rarely occurs worldwide, and has not been reported in Korea. We present here a case of ophthalmomyiasis caused by Phormia sp. fly larva in an enucleated eye of a patient. In June 2010, a 50-year-old man was admitted to Dankook University Hospital for surgical excision of a malignant melanoma located in the right auricular area. He had a clinical history of enucleation of his right eye due to squamous cell carcinoma 5 years ago. During hospitalization, foreign body sensation developed in his right eye, and close examination revealed a fly larva inside the eye, which was evacuated. The larva was proved to be Phormia sp. based on the morphology of the posterior spiracle. Subsequently, no larva was found, and the postoperative course was uneventful without any complaints of further myiasis. This is the first case of ophthalmomyiasis among the literature in Korea, and also the first myiasis case caused by Phormia sp. in Korea.     

Beyond barcoding: A mitochondrial genomics approach to molecular phylogenetics and diagnostics of blowflies (Diptera: Calliphoridae)

Members of the Calliphoridae (blowflies) are significant for medical and veterinary management, due to the ability of some species to consume living flesh as larvae, and for forensic investigations due to the ability of others to develop in corpses. Due to the difficulty of accurately identifying larval blowflies to species there is a need for DNA-based diagnostics for this family, however the widely used DNA-barcoding marker, cox1, has been shown to fail for several groups within this family. Additionally, many phylogenetic relationships within the Calliphoridae are still unresolved, particularly deeper level relationships. Sequencing whole mt genomes has been demonstrated both as an effective method for identifying the most informative diagnostic markers and for resolving phylogenetic relationships. Twenty-seven complete, or nearly so, mt genomes were sequenced representing 13 species, seven genera and four calliphorid subfamilies and a member of the related family Tachinidae. PCR and sequencing primers developed for sequencing one calliphorid species could be reused to sequence related species within the same superfamily with success rates ranging from 61% to 100%, demonstrating the speed and efficiency with which an mt genome dataset can be assembled. Comparison of molecular divergences for each of the 13 protein-coding genes and 2 ribosomal RNA genes, at a range of taxonomic scales identified novel targets for developing as diagnostic markers which were 117–200% more variable than the markers which have been used previously in calliphorids. Phylogenetic analysis of whole mt genome sequences resulted in much stronger support for family and subfamily-level relationships. The Calliphoridae are polyphyletic, with the Polleninae more closely related to the Tachinidae, and the Sarcophagidae are the sister group of the remaining calliphorids. Within the Calliphoridae, there was strong support for the monophyly of the Chrysomyinae and Luciliinae and for the sister-grouping of Luciliinae with Calliphorinae. Relationships within Chrysomya were not well resolved. Whole mt genome data, supported the previously demonstrated paraphyly of Lucilia cuprina with respect to L. sericata and allowed us to conclude that it is due to hybrid introgression prior to the last common ancestor of modern sericata populations, rather than due to recent hybridisation, nuclear pseudogenes or incomplete lineage sorting.     

Gastrointestinal myiasis by larvae of Sarcophaga sp. and Oestrus sp. in Egypt: report of cases, and endoscopical and morphological studies

Four cases of gastric or intestinal myiasis are reported. The cases contain 2 males (1 child 10 years old, and 1 adult 40 years old) and 2 females (1 girl 18 years old, and 1 adult 50 years old) from Minia Governorate, Southern Egypt. Three of them, including cases no. 1, 3, and 4, were gastric myiasis, and complained of offensive hematemesis of bright red blood. Minute moving worms, larvae of the fly, were found in the vomitus. On the other hand, case no. 2 had intestinal myiasis, and complained of abdominal distention, nausea, vomiting, and diarrhea. The stool of case 2 was mixed with blood, and minute moving worms were observed in the stool. Endoscopy was performed to explore any pathological changes in the stomach of the patients. The larvae were collected and studied macroscopically, microscopically, and us-ing a scanning electron microscope (SEM) to identify their species. Three different types of larvae were identified. The larvae isolated from case 1 were diagnosed as the second stage larvae of Sarcophaga species, and the larvae isolated from case 2 were the third stage larvae of Sarcophaga species. On the other hand, the larvae isolated from cases 3 and 4 were diagnosed as the third stage larvae of Oestrus species.     

Morphological and Ecological Characterization of Steinernema feltiae (Rhabditida: Steinernematidae) Rioja Strain Isolated from Bibio hortulanus (Diptera: Bibionidae) in Spain

A new strain of Steinernema feltiae (Rhabditida: Steinernematidae) was isolated in La Rioja (Spain) from larvae of Bibio hortulanus (Diptera: Bibionidae). A comparative morphometric analysis of this new strain and four additional S. feltiae isolates was performed. Although significant differences in morphometric measurements were observed, PCR-RFLP profiles and sequence analysis of the ITS region of rDNA confirmed the identity of the new strain as A2 RFLP type of S. feltiae. A comparative morphometric study among nematodes from three hosts, Galleria mellonella (Lepidoptera: Pyralidae), Spodoptera littoralis (Lepidoptera: Noctuidae) and B. hortulanus, was conducted. Ecological characterization of the Rioja isolate was performed in G. mellonella larvae. Larval mortality was 75.3 and 78.12% in penetration and sand column assays, respectively, and the percentage of penetrating infective juveniles was 12.0 and 2.8% in these assays. Larval mortality in the one-on-one bioassay was 4.2%, and in exposure-time bioassays, it was 50% at 11.25 hours. Relationships between morphometric characteristics and host mortality are discussed for this new strain of entomopathogenic nematode.     

Human urogenital myiasis caused by the ‘rat-tailed’ larvae of Palpada scutellaris (Fabricius, 1805) in Santander,eastern Colombia: A case report

The Palpada genus, which belongs to the Diptera order (family, Syrphidae), has been rarely reported to cause accidental myiasis in humans. Herein, we report the first case of genitourinary myiasis caused by a larva of the Palpada genus in a 9-year-old girl from Colombia. The girl, who resided in a rural area in the municipality of Floridablanca, Santander, near Bucaramanga city, in eastern Colombia, presented with lower abdominal pain accompanied by oliguria, followed by the subsequent elimination of a larva through the urine. The next day, the patient visited a primary healthcare centre, and no signs or symptoms were observed on clinical examination. Haematological analysis showed high plateletcrit levels and platelet large cell counts. The results of the urine test revealed a decrease in specific gravity and a slight increase in bacterial content and mucus. DNA barcoding analyses showed that the etiological agent corresponded to a third instar larva of the Palpada scutellaris species. This is the first case to report genitourinary myiasis caused by larvae of the genus Palpada in humans. However, we believe that additional cases might be accurately detected if adequate tests are performed to confirm the clinical and molecular features associated with this infection.     

A Nasal Myiasis in a 76-Year-Old Female in Korea

On July 2009, 5 fly larvae were discovered inside the nose of a 76-year-old female. She was living in Cheonan-si, and in a state of coma due to rupture of an aortic aneurysm. Surgery was performed on the day of admission, and the larvae were found 4 days later. By observing their posterior spiracle, the larvae were identified as Lucilia sericata. Considering the rapid development of this species, the infection was likely acquired during hospitalization. Further investigation on the hospital environment should be needed to know the origin of the infection.     

Species Diversity and Seasonal Distribution of Culicoides spp. (Diptera: Ceratopogonidae) in Jeju-do,Republic of Korea

Biting midges belonging to the genus Culicoides (Diptera: Ceratopogonidae) were collected by Mosquito Magnet^® and black light traps at 5 sites on Jeju-do, Republic of Korea (Korea), from May-November 2013 to determine species diversity and seasonal distribution. A total of 4,267 specimens were collected, of which 99.9% were female. The most common species was Culicoides tainanus (91.8%), followed by C. lungchiensis (7.2%) and C. punctatus (0.6%), while the remaining 4 species accounted for <0.5% of all Culicoides spp. that were collected. High numbers of C. tainanus were collected in May, followed by decreasing numbers through August, and then increasing numbers through November when surveillance was terminated. Peak numbers of C. lungchiensis were collected during September, with low numbers collected from May-August and October-November. The presence of C. lungchiensis in Korea was confirmed by morphological and molecular analyses.     

The toad fly Lucilia bufonivora: its evolutionary status and molecular identification

The blow fly genus Lucilia is composed largely of saprophages and facultative myasis agents, including the economically important species Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae) and Lucilia sericata (Meigen). Only one species is generally recognized as an obligate agent of myiasis, Lucilia bufonivora Moniez, and this is an obligate parasite of toads. Lucilia silvarum (Meigen), a sister species, behaves mainly as a carrion breeder; however, it has also been reported as a facultative parasite of amphibians. Morphologically, these species are almost identical, and historically this has led to misidentification, taxonomic ambiguity and a paucity of studies of L. bufonivora. In this study, dipterous larvae were analysed from toad myiasis cases from the U.K., The Netherlands and Switzerland, together with adult specimens of fly species implicated in amphibian parasitism: L. bufonivora, L. silvarum and Lucilia elongata Shannon (from North America). Partial sequences of two genes, cox1 and ef1α, were amplified. Seven additional blow fly species were analysed as outgroups. Bayesian inference trees of cox1, ef1α and a combined‐gene dataset were constructed. All larvae isolated from toads were identified as L. bufonivora and no specimens of L. silvarum were implicated in amphibian myiasis. This study confirms L. silvarum and L. bufonivora as distinct sister species and provides unambiguous molecular identification of L. bufonivora.     

Analysis of somatic and salivary gland antigens of third stage larvae of Rhinoestrus spp. (Diptera, Oestridae)

Larvae of Rhinoestrus spp. (Diptera, Oestridae) infect nasal and sinus cavities of horses, causing a nasal myiasis characterized by severe respiratory distress. Presently, the diagnosis of horse nasal botfly relies on the observation of clinical signs, on the post mortem retrieval of larvae or on molecular assays performed using pharyngeal swabs. The present study was carried out to characterize larval somatic proteins and salivary glands of Rhinoestrus spp. in a preliminary assessment towards the immunodiagnosis of equine rhinoestrosis. Out of the 212 necropsied horses 13 were positive for the presence of Rhinoestrus spp. larvae. The analysis of the sera from the infected animals by Western blotting assay showed the presence of a specific host humoral immune response against Rhinoestrus spp. larvae and proved that the salivary glands are the major immunogens in horse nasal botflies.     

Characterization of the Cystoid Nematode Meloidoderita kirjanovae (Nemata: Sphaeronematidae) from Southern Italy

A population of the cystoid nematode Meloidoderita kirjanovae was detected parasitizing water mint (Mentha aquatica) in southern Italy. The morphological identification of this species was confirmed by molecular analysis using the internal transcribed spacer 1 (ITS1) and 5.8S gene sequences of nuclear ribosomal DNA (rDNA), which clearly separated it from the closely related species Meloidoderita polygoni. A phylogenetic analysis of M. kirjanovae with species of related genera was conducted using sequences of the D2-D3 expansion segments of the 28S nuclear ribosomal RNA gene. The resulting phylogenetic tree was congruent with trees from an extended dataset for Criconematina and Tylenchida. The basal position of the genus Meloidoderita together with Sphaeronema within the Criconematina clade in this tree may indicate their close relationships. The anatomical changes induced by M. kirjanovae population from Italy in water mint were similar to those reported for a nematode population infecting roots of M. longifolia in Israel. Nematode feeding caused the formation of a stellar syncytium that disorganized the pericycle and vascular root tissues.     

Molecular Variation and Distribution of Anopheles fluviatilis (Diptera: Culicidae) Complex in Iran

Anopheles fluviatilis James (Diptera: Culicidae) is one of the known malaria vectors in south and southeastern Iran. Earlier ITS2 sequences analysis of specimens from Iran demonstrated only a single genotype that was identical to species Y in India, which is also the same as species T. We identified 2 haplotypes in the An. fluviatilis populations of Iran based on differences in nucleotide sequences of D3 domain of the 28S locus of ribosomal DNA (rDNA). Comparison of sequence data from 44 Iranian specimens with those publicly available in the Genbank database showed that all of the 28S-D3 sequences from Kazeroun and Khesht regions in Fars Province were identical to the database entry representing species U in India. In other regions, all the individuals showed heterozygosity at the single nucleotide position, which identifies species U and T. It is argued that the 2 species may co-occur in some regions and hybridize; however, the heterozygosity in the 28S-D3 locus was not reflected in ITS2 sequences and this locus for all individuals was identical to species T. This study shows that in a newly diverged species, like members of An. fluviatilis complex, a single molecular marker may not be sufficiently discriminatory to identify all the taxa over a vast geographical area. In addition, other molecular markers may provide more reliable information for species discrimination.     

Three new species of European Platypalpus (Diptera,Hybotidae)

Platypalpus graecoides sp. n. (Italy), Platypalpus pyreneensis sp. n. (Andorra), and Platypalpus silvahumidus sp. n. (Czech Republic) are described. All three species are illustrated and keyed. Platypalpus hallensis Grootaert & Stark, 1997 is first reported from France and Spain.