Transcriptomic profiling of different developmental stages reveals parasitic strategies of Wohlfahrtia magnifica, a myiasis-causing flesh fly.

BACKGROUND: Wohlfahrtia magnifica is an obligatory parasite that causes myiasis in several warm-blooded vertebrates. Adult females deposit the first-stage larvae directly onto wounds or natural body orifices (e.g., genitalia) of the host, from where they quickly colonize the host tissue and feed on it for development. The infestation of W. magnifica can lead to health issues, welfare concerns, and substantial economic losses. To date, little is known about the molecular mechanisms of the W. magnifica-causing myiasis. RESULTS: In this study, we collected parasitic-stage larvae of W. magnifica from wounds of naturally infested Bactrian camels, as well as pupae and adult flies reared in vitro from the wound-collected larvae, for investigating the gene expression profiles of the different developmental stages of W. magnifica, with a particular focus on examining gene families closely related to the parasitism of the wound-collected larvae. As key proteins related to the parasite-host interaction, 2049 excretory/secretory (ES) proteins were identified in W. magnifica through the integration of multiple bioinformatics approaches. Functional analysis indicates that these ES proteins are primarily involved in cuticle development, peptidase activity, immune response, and metabolic processes. The global investigation of gene expression at different developmental stages using pairwise comparisons and weighted correlation network analysis (WGCNA) showed that the upregulated genes during second-stage larvae were related to cuticle development, peptidase activity, and RNA transcription and translation; during third-stage larvae to peptidase inhibitor activity and nutrient reservoir activity; during pupae to cell and tissue morphogenesis and cell and tissue development; and during adult flies to signal perception, many of them involved in light perception, and adult behavior, e.g., feeding, mating, and locomotion. Specifically, the expression level analysis of the likely parasitism-related genes in parasitic wound-collected larvae revealed a significant upregulation of 88 peptidase genes (including 47 serine peptidase genes), 110 cuticle protein genes, and 21 heat shock protein (hsp) genes. Interestingly, the expression of 2 antimicrobial peptide (AMP) genes, including 1 defensin and 1 diptericin, was also upregulated in the parasitic larvae. CONCLUSIONS: We identified ES proteins in W. magnifica and investigated their functional distribution. In addition, gene expression profiles at different developmental stages of W. magnifica were examined. Specifically, we focused on gene families closely related to parasitism of wound-collected larvae. These findings shed light on the molecular mechanisms underlying the life cycle of the myiasis-causing fly, especially during the parasitic larval stages, and provide guidance for the development of control measures against W. magnifica.

Psychodid flies and their implicated role in human myiasis and pseudomyiasis.

Several psychodid flies are commonly associated with human-inhabited environments and have been increasingly implicated in cases of human myiasis. However, the basic biology of psychodid larvae is not well-suited for survival in the human intestinal or urogenital tract, making true, prolonged myiasis unlikely. In this review, we performed a systematic literature review of published cases of purported myiasis caused by psychodid flies, their identification, associated clinical findings, and treatment. We also discuss the anatomy and lifecycle of psychodid flies in relation to their purported ability to use human tissue as a nutritive source and survive in the human alimentary or urogenital tracts. Based on the range of non-specific and varied reported clinical manifestations, lack of observed collections, life cycle patterns of psychodid flies, the mechanics of their mouthparts, and breathing requirements, we conclude that most cases likely represent incidental findings, or in rare cases possibly pseudomyiasis, rather than true myiasis, and provide recommendations for clinical evaluation and reporting so that disease misclassification and unnecessary therapy do not occur.

Bot fly parasitism in mantled howler monkeys (Alouatta palliata): General patterns and climate influences.

Parasitism is a strong selective pressure, and its study is crucial for predicting the persistence of host species. Mantled howler monkeys are infected by the larvae of the bot fly Cuterebra baeri. This parasitosis produces myiasis and may have negative impacts on host health, although systematic information on the dynamics of this host-parasite relationship is very limited. Currently, all available information on infection patterns of C. baeri comes from a single mantled howler monkey population (Barro Colorado Island, Panama). Therefore, in this study we describe temporal variation in infection patterns for a newly mantled howler monkey population and analyze the relationship between climate and infection likelihood. We assessed the presence of C. baeri nodules in 17 adult individuals in Los Tuxtlas for 10 months through direct observation and compiled data on ambient temperature and rainfall. Most subjects had nodules during the study and there were no differences between sexes in the number of nodules. Nodules were usually located in the neck. Prevalence and abundance of nodules peaked thrice during the study (February, April, and September), a pattern that was very similar to that of parasitism intensity (February, April, and August). Incidence closely tracked these peaks, increasing before and decreasing after them. The likelihood of nodule appearance increased when both mean and minimum temperature decreased in the 24-21 prior days to nodule appearance. It also increased with decreased rainfall in the 5-2 prior days to nodule appearance. Although only three of the eight analyzed climate variables had a significant effect on parasitosis, these results suggest that climate may affect pupal development and the access of larvae to hosts. Besides contributing data on C. baeri parasitism for a new mantled howler monkey population, our study provides novel information on the influence of environmental factors on the dynamics of host-parasite systems.

Ophthalmomyiasis Case Caused by Two Blow Fly (Diptera: Calliphoridae) Species in North America.

Ophthalmomyiasis is the result of fly larvae feeding on the tissues of the eye. Commonly associated with poor hygiene and open wounds, this condition is rare and often stigmatized. Treatment can be straightforward, and full recovery is common. Identifying the species responsible for ophthalmomyiasis is important for the medical, forensic, and entomological communities. Here, we present a case of ophthalmomyiasis where 30-40 blow fly (Diptera: Calliphoridae) larvae were removed from the eye of a human male. A representative subsample of five larvae was used for taxonomic identification via two approaches (a) DNA analysis, via sequencing of the complete mitochondrial genome (mtGenome) and comparison of the mtGenome and mitochondrial COI barcode region to GenBank, and (b) morphology, examination of the posterior spiracles using microscopy, and comparison to published larval descriptions of blow flies. Two species of blow flies were identified from the DNA analysis: Lucilia coeruleiviridis and Phormia regina. Morphological examination could only confirm L. coeruleiviridis as being present. To our knowledge, finding two blow fly species causing ophthalmomyiasis in a single individual has not been previously reported in the scientific literature. Neither P. regina nor L. coeruleiviridis prefers living tissue for larva development, but since they fill similar ecological niches, perhaps this was a show of competition rather than a normal feeding habit. Knowing these blow fly species can resort to this behavior, and that it can affect human populations, is valuable to the education of patients and providers.

Larval anatomy of the digestive and excretory systems of the pharyngeal bot fly, Pharyngomyia picta (Diptera: Oestridae).

Oestrid flies (Diptera: Oestridae) are obligate parasites of mammals during their larval stage and show anatomical adaptations for the infestation of host tissues. Unlike the species that parasitize domestic mammals, those oestrid species that infest wild mammal hosts remain poorly known. With the use of x-ray micro-computed tomography, we describe for the first time the anatomy of the digestive and excretory systems of the second and third larval instars of Pharyngomyia picta (Meigen), a parasite of cervids that, like other species within the subfamily Oestrinae, causes nasopharyngeal myiasis. Both larval instars of P. picta show a pair of remarkably large salivary glands arranged in a characteristic 'glandular band', a convoluted and thickly uniform midgut and a greatly enlarged distal region of the anterior pair of Malpighian tubules. These anatomical features also have been described in other species within the subfamily Oestrinae, whereas they differ from the observations in other oestrid subfamilies. We discuss the potential functional significance of the anatomy of the digestive and excretory systems of Oestrinae larvae as specific adaptations to parasitize the nasopharyngeal cavities of mammal hosts.

Larval morphology of the avian parasitic genus Passeromyia: playing hide and seek with a parastomal bar.

The enigmatic larvae of the Old World genus Passeromyia Rodhain & Villeneuve, 1915 (Diptera: Muscidae) inhabit the nests of birds as saprophages or as haematophagous agents of myiasis among nestlings. Using light microscopy, confocal laser scanning microscopy and scanning electron microscopy, we provide the first morphological descriptions of the first, second and third instar of P. longicornis (Macquart, 1851) (Diptera: Muscidae), the first and third instar of P. indecora (Walker, 1858) (Diptera: Muscidae), and we revise the larval morphology of P. heterochaeta (Villenueve, 1915) (Diptera: Muscidae) and P. steini Pont, 1970 (Diptera: Muscidae). We provide a key to the third instar of examined species (excluding P. steini and P. veitchi Bezzi, 1928 (Diptera: Muscidae)). Examination of the cephaloskeleton revealed paired rod-like sclerites, named 'rami', between the lateral arms of the intermediate sclerite in the second and third instar larva. We reveal parastomal bars fused apically with the intermediate sclerite, the absence of which has so far been considered as apomorphic for second and third instar muscid larvae. Examination of additional material suggests that modified parastomal bars are not exclusive features of Passeromyia but occur widespread in the Muscidae, and rami may occur widespread in the Cyclorrhapha.

Prevalence of Oestrus ovis in small ruminants from the eastern Iberian Peninsula. A long-term study.

Oestrus ovis is an obligate parasite that causes myiasis in domestic ruminants, being commonly found in the Mediterranean area. From 2009 to 2019 a total of 3476 heads of culling sheep and goats from the Mediterranean coast of Spain were examined for the presence of O. ovis. The total prevalence was 56.3%, significantly higher in sheep than in goats (61.2% and 43%, respectively). Differences were found in the mean annual prevalence, with the highest value being registered in 2018 (61.7%) and the lowest in 2012 (50.3%). Autumn, for sheep, and winter, for goats, were the seasons with the highest number of infested specimens. Temperature, but not rainfall, was found to be associated with prevalence (p < 0.05). Most L1 were found in the anatomic region I (septum, meatus, and ventral conchae), while L2 and L3 were mainly located in regions II (nasopharynx, ethmoid labyrinth, and dorsal conchae), and III (sinuses). The overall intensity was 12.8 larvae per head, significantly higher in sheep (13.3) than in goats (3.5). Our results confirm the high prevalence of O. ovis in sheep and goats in this geographic area over the last decade, with the trend increasing in recent years in association with higher mean temperatures.

An Atypical Presentation Of Cutaneous Myiasis Of The Scalp In North Punjab: A Case Report.

Myiasis is an ectoparasitic infestation caused by larvae of arthropods in the group Diptera. Cutaneous myiasis is the infection of skin and includes the following types: furuncular, migratory and wound myiasis. Out of all the organisms responsible for this disease, Wohlfahrtia vigil is the most common causative agent in Pakistan, usually seen from June to September with nearly all presentations occurring at a young age. In our case, a patient without a history of trauma, skin erosion or animal exposure has a unique presentation of cutaneous myiasis. Here, an effort was made to treat a 15- year-old otherwise healthy female with a multisegmented treatment modality. A follow up visit was conducted every three days for three months at the end of which a 60% decrease in the size of the open wound was observed.

Oral myiasis in humans.

Myiasis is a parasitic infestation resulting from flies laying eggs in the host tissues. It is common in animals, but can also occur in humans, including in the oral cavity. The diagnosis is usually quite clear in endemic regions such as Brazil, but it can be challenging to clinicians in nonendemic counties when faced with a patient who has acquired the infection elsewhere. We report two clinical cases of human intraoral myiasis and discuss the diagnosis and treatment of this condition. Two men, both of whom were in situations of vulnerability presented with myiasis: the first patient had larvae infesting the alveolar ridge region after tooth extraction, while the second was diagnosed with inflammatory fibrous hyperplasia associated with larvae along the edges of the lesion. Both were treated with ivermectin and antibiotics, and given guidance on preventative care. Such infestations are often a clear sign of neglect, and clinicians need to pay attention to the general health of patients affected by oral myiasis.

Evolution of coding sequence and gene expression of blowflies and botflies with contrasting feeding habits.

The Oestroidea superfamily is characterized by the diversity of feeding preferences among closely-related species; these flies are saprophagous, obligate parasites, or facultative parasites. We used gene expression and coding sequence data from five species (Cochliomyia hominivorax, Chrysomya megacephala, Lucilia cuprina, Dermatobia hominis, and Oestrus ovis) to identify underlying genetic differences involved in the diverse lifestyles. We tested whether 1287 orthologs have different expression and evolutionary constraints under different scenarios. We found two up-regulated genes; one in species causing cutaneous myiasis that is involved in iron transportation/metabolization (ferritin), and another in species causing traumatic myiasis that responds to reduced oxygen levels (anoxia up-regulated-like). Our evolutionary analysis showed a similar result. In the Co. hominivorax branch, we found one gene with the same function as ferritin that may be evolving under positive selection, spook. This is the first step towards understanding origins and evolution of parasitic strategy diversity in Oestroidea.

Point-of-Care Ultrasound Findings in a Case of Botfly Myiasis Contracted in the United States.

Botfly infiltration is a rare cause of pediatric skin manifestations in the United States, but should be considered in nonhealing wounds even in nontravelers. We describe the case of a healthy 6-y-old female who had never traveled outside of the southeast United States, presenting with a nonhealing skin lesion. The point-of-care ultrasound (POCUS) findings suggested subcutaneous parasitic infiltration. This case demonstrates the role of POCUS in identification of subcutaneous parasitic infiltration, and differentiation from other, more common skin lesions.

Prevalence of tracheopulmonary myiasis amidst humans.

Myiasis is a parasitic infestation of live vertebrates with the dipteran larvae that grow within the host while feeding on its living or dead tissue. Human myiasis is prevalent among individuals in close association of domestic animals and those inhabiting the unhygienic conditions. Open neglected suppurative wounds are the favourable sites that attract and stimulate the gravid female flies for oviposition. However, the poor personal hygiene, ignorance and the mental illness further add to it. Tracheopulmonary myiasis involves the infestation of foul smelling nasal and oral orifices or neglected wounds like tracheostomal incisions with the parasitic dipteran larvae which may extend up to the trachea, bronchi and lungs leading to serious health complications. Despite the incidence of human tracheopulmonary infestation is rare, the severe medical impediments and heterogeneous worldwide distribution signify the medical importance of this condition. The present manuscript reviews and summarizes the worldwide reported cases of human tracheopulmonary myiasis and the predisposing risk factors for onset of the same, as future reference for parasitologists and medical professionals.

Effect of a novel juvenoid fenoxycarb on the pupal-adult transformation in the blowfly, Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae).

Chrysomya megacephala (Fabricius, 1794) (Diptera, Calliphoridae) acts as a vector of many disease-causing pathogens. It causes myiasis in human beings and other living vertebrates. In the present study, the effect of a juvenile hormone analog (JHA), fenoxycarb, was evaluated on pupal-adult transformation by exposing pupae (0 and 1-day-old) of blowfly. Pupae were topically treated with different concentrations of the compound, viz., 20, 40, and 80 µg/µl applied on the posterior dorsum with the help of a micropipette. The effects comprised various developmental aberrations, such as delayed pupal-adult ecdysis duration, pupal mortality, formation of pupal-adult intermediates, eclosion failure, reduced adult emergence, and formation of abnormal adults. The freshly molted pupae (0 day) were more susceptible as compared to 1-day-old pupae. Pupal mortality was highest at 80 µg. Normal adult emergence was completely inhibited at day 0 pupae treated with 80 µg of the fenoxycarb. These results demonstrate that fenoxycarb was capable of successfully inhibiting the pupal-adult transformation, and thus, it can be used to control this myiasis-causing agent.

Ignatzschineria larvae Bacteremia Following Lucilia sp. Myiasis in an Irregular Migrant: A Case Report.

Bacteremia induced by wound myiasis is uncommon and therefore rarely suspected by clinicians when treating patients with neglected wounds. We present a case of Ignatzschineria larvae bacteremia as a complication of Lucilia sp. maggot wound myiasis in a young male migrant. This is the first reported human case of Ignatzschineria bacteremia in Slovenia and one of the 2 described in the literature where the fly larvae infesting the wounds of the patient with Ignatzschineria bacteremia were not only suspected to be Lucilia sp. but also entomologically identified.

In vitro test for the evaluation of the efficacy of topical products for the control of Cochliomyia hominivorax larvae.

Cochliomyia hominivorax larvae cause myiasis in animals and humans. To register a commercial product to control this dipteran is necessary to experiment on animals. The in vitro test was standardized to evaluate the larvicidal efficacy of commercial topical products. Five formulations were analysed in vitro and in vivo. For the in vitro test, a colony was formed and three replicates (n = 200) of each larval stage (L1, L2 and L3) were treated. The viability of the larvae was evaluated after 5 and 30 min, and at 1, 2, 6, 12, 24, 48, 60 and 72 h post-treatment (HPT). For the in vivo test, 30 bovines divided into six groups were castrated to achieve natural infestation with C. hominivorax. Animals in the treated groups received the product. Myiasis and efficacy were evaluated 12, 24, 36, 48, 60 and 72 HPT. Four formulations tested in the in vitro test achieved 100% efficacy at 24 HPT. In the in vivo experiment only one achieved 100% efficacy at 24 HPT. However, all products achieved the maximum efficacy by the end of study. The in vitro test developed here could be adopted to evaluate the efficacy of topical products for the control of C. hominivorax larvae.

Furuncular myiasis of the scalp associated with pigmented basal cell carcinoma: First case diagnosed in Albania.

We are reporting a case of myiasis associated with pigmented basal cell carcinoma. This is the first reported case from Albania and Balkan peninsula. Correct diagnosis and prompt treatment may prevent a disastrous and destructive course of the disease and may lead to improved quality of life.

Deer keds on wild ungulates in northern Italy, with a taxonomic key for the identification of Lipoptena spp. of Europe.

Deer keds (Lipoptena spp.) are blood-sucking ectoparasites of domestic and wild animals, and also accidentally of humans. In Europe, five Lipoptena spp. have been recorded, although the lack of specific taxonomic keys has often led to mistaken identification or to missing data. The present study aimed to develop an identification key of the European species and also to identify Lipoptena spp. found on wild ungulates in northern Italy. In total, 390 hippoboscids were collected from Rupicapra rupicapra, Capreolus capreolus, Cervus elaphus and Ovis aries musimon in an Alpine area of Italy. After morphological identification, 140 specimens were subjected to phylogenetic analysis based on mitochondrial (CO1) and nuclear (CAD) gene sequences. Despite the expected presence of slight morphological variations, all specimens examined were identified both microscopically and molecularly as Lipoptena cervi (100% identity for both CO1 and CAD genes). The massive increase in wild ungulate populations can favour the possibility of detecting other species of Lipoptena. The identification keys proposed in the present study may help with monitoring the presence of Lipoptena species, particularly in European countries where this ectoparasite is neglected and for which various data (from diffusion to control methods) are still missing.

Development and characterization of 15 novel polymorphic microsatellite loci for two important bot flies (Diptera, Oestridae) by next-generation sequencing.

Cephenemyia stimulator and Oestrus ovis are two important parasitic bot flies (Oestridae) species causing myiasis, with a potential negative impact on the welfare of the host. Using next-generation sequencing approach and bioinformatics tools, a large panel of possible microsatellites loci was obtained in both species. Primer pairs were designed for 15 selected microsatellite loci in C. stimulator and other 15 loci in O. ovis for PCR amplification. Loci amplification and analysis were performed in four populations of each species. The results demonstrated that all selected loci were polymorphic, with the number of alleles ranging from 2 to 6 per locus in C. stimulator and 3 to 13 per locus in O. ovis. This is the first time to describe these microsatellite loci for C. stimulator and O. ovis. These two sets of microsatellite markers could be further used for biogeographic and population genetics studies.