Molecular identification of Chrysomya albiceps (Diptera: Calliphoridae) in a case of primary cutaneous myiasis in sheep at southern Brazil.

Sheep farming has been growing in Brazil, driven by an expanding consumer market due to greater acceptance of its meat and derivatives. There are several factors that limit sheep production, and one of them is infestation by ectoparasites, which cause stress in animals, weight loss, poor development, low productivity, low quality wool and reduced fertility. Chrysomya albiceps is a species of blowfly belonging to the Calliphoridae family that occurs in neotropical regions, where it causes secondary myiasis. We identified here a rare case of cutaneous myiasis with the presence of tissue lesions caused by C. albiceps in sheep in southern Brazil. We highlight the need to carry out more in-depth studies regarding the biology of these insects, with the aim of proving this atypical behavior for Brazil.

Prevalence and molecular identification of hypodermosis from slaughtered cattle in Sulaymaniyah province, Iraq.

INTRODUCTION: Hypodermosis is a subcutaneous infestation in cattle that is caused by larvae of Hypoderma spp. and it is an economically important disease in the cattle industry. This study aimed to find the prevalence rate of hypodermosis and identify variations in the COX1 gene among isolates present in Sulaymaniyah, in the Kurdistan region of Iraq. METHODOLOGY: The study was conducted in a Sulaymaniyah slaughterhouse from March to July 2021. The carcasses of 867 cattle were carefully checked before and after skinning them to record the presence of boils containing the larvae of Hypoderma spp. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using TaqI enzyme, and sequencing of the COX1 gene were used for diagnosis and molecular characterization of Hypoderma spp. RESULTS: The rate of infestation with Hypoderma bovis was 1.61% and the highest rate (3.57%) was detected in April. The disease was significantly (p < 0.05) higher in local breeds at 2.79%. PCR-RFLP confirmed that all recorded species were H. bovis. The result was further confirmed by Sanger sequencing of the COX1 gene of the isolated species. Comparison of the sequences of the conserved COX1 gene of the parasite led to identification of six different haplotypes in the research area. Two of the haplotypes were previously recorded internationally, while four new haplotypes associated with four novel mutations were recorded for the first time in the study region. CONCLUSIONS: Based on these results we can conclude that H. bovis is a widespread species in the research region.

Effectiveness of lotilaner on furuncular myiasis in dogs naturally infested with Dermatobia hominis (Diptera: Cuterebridae).

An evaluation was made of the larvicidal efficacy of lotilaner (Credeli®) in the treatment of dogs naturally infested with Dermatobia hominis larvae. A total of 12 dogs presenting at least three live D. hominis larvae were medicated. The animals were medicated orally with a single dose of no less than 20 mg/kg lotilaner. After drug administration, the animals remained at their homes, and observations were made to verify the larvicidal effect 6 hours after treatment. Live larvae were considered any parasite that exhibited motility after removal. For each animal was using the formula: 100 x [(total of live larvae before treatment - total live larvae after treatment) /total of live larvae before treatment] as criteria for evaluating lotilaner efficacy. A total of 98 larvae were counted in 12 dogs, with an average of 8.1 larvae per animal. The effectiveness of lotilaner was 80.6%. Nineteen larvae were found alive, albeit presenting hypomobility and lethargic behavior. However, note that the evaluation was performed just six hours after administration of the drug. Lotilaner administered orally in a single dose of 20 mg/kg showed 80.6% efficacy six hours after treating dogs naturally infested with D. hominis.

Surgical management for the case of scrotal myiasis in a 7-day-old neonate: a case report.

INTRODUCTION: Neonatal myiasis is a rare condition, with few reports available on the subject. Surgical management is recommended in some cases. In this study, we present the case of a 7-day-old male neonate with larvae in his scrotum who underwent surgery. CASE PRESENTATION: A full-term 7-day-old male infant (Aryan race) was referred to a children's hospital. On the sixth day after birth, three 3-4 mm long larvae crawled out from his scrotum, with the number increasing over time. He was given intravenous antibiotics and topical mupirocin to combat secondary infections. The surgical treatment involved two steps: first, the larvae were extracted, and then the infection site was washed with betadine and hydrogen peroxide to help remove any possible remaining larvae. CONCLUSION: Scrotal myiasis is a rare disease that occurs in infants and requires immediate treatment. Surgical treatment is effective in removing dead or decaying larvae from a deep-seated location and washing the infection site to prevent secondary infection.

Gasterophilus intestinalis infestation in lion (Panthera leo) and plains zebra (Equus quagga) in the Serengeti ecosystem: Morphological and molecular profiling.

This study was conducted to clarify the host specificity and the geographical distribution of Gasterophilus species (Diptera, Oestridae) in the Serengeti ecosystem. A total of 317 larvae were recovered from two common zebras (Equus quagga, formerly Equus burchellii) in Maswa Game Reserve, and 58 larvae were recovered from an African lion (Panthera leo) in the Serengeti National Park. The study emphasizes the rare occurrence of Gasterophilus sp. in lions, shedding light on the broader life cycle and physiological implications for hosts. Genetic analysis of cox2 genes from Gasterophilus species, sourced from a single geographic location, reveals significant genetic distinctions and host specificity. This study reports the first case of G. intestinalis infestation in an African lion in the Serengeti ecosystem, extending its known range from zebras and other equids, and highlighting ecological and veterinary implications. This unusual prey-predator transmission highlights the value of molecular taxonomic tools in clarifying host-parasite dynamics and guiding targeted conservation strategies.

Title: Infestation par Gasterophilus intestinalis chez le lion (Panthera leo) et le zèbre des plaines (Equus quagga) dans l'écosystème du Serengeti : profilage morphologique et moléculaire. Abstract: Cette étude a été menée pour clarifier la spécificité de l'hôte et la répartition géographique des espèces de Gasterophilus (Diptera, Oestridae) dans l'écosystème du Serengeti. Au total, 317 larves ont été récoltées chez deux zèbres communs (Equus quagga, anciennement Equus burchellii) dans la réserve de gibier de Maswa, et 58 larves ont été récoltées chez un lion d'Afrique (Panthera leo) dans le parc national du Serengeti. L'étude souligne la rareté de l'occurrence de Gasterophilus sp. chez les lions, mettant en lumière le cycle biologique plus large et les implications physiologiques pour les hôtes. L'analyse génétique des gènes cox2 des espèces de Gasterophilus, provenant d'un seul lieu géographique, révèle des distinctions génétiques et une spécificité d'hôte significatives. Cette étude rapporte le premier cas d'infestation par G. intestinalis chez un lion africain dans l'écosystème du Serengeti, étendant son aire de répartition déjà connue chez les zèbres et autres équidés, et mettant en évidence des implications écologiques et vétérinaires. Cette transmission inhabituelle de proie à prédateur souligne l'intérêt des outils de taxonomie moléculaire pour clarifier la dynamique hôte-parasite et guider les stratégies de conservation ciblées.

Quantitative proteomics reveals significant variation in host responses of cattle with differing buffalo fly susceptibility.

Background: Control of buffalo flies (Haematobia irritans exigua, BFs) relies mainly on chemical methods; however, resistance to insecticides is widespread in BF populations. Breeding for resistance to BFs represents a possible alternative, but direct phenotyping of animals is laborious and often inaccurate. The availability of reliable diagnostic biomarker(s) to identify low BF carrier cattle would facilitate rapid and accurate selection for genetic improvement. However, limited information is available regarding differences amongst cattle in host responses to BF infestation. Methods: This study investigated the variation in Brangus cattle serum proteomic profiles before (naïve) and after peak BF exposure, in low (LF) and high BF burden (HF) cattle. Cattle were phenotyped for susceptibility based on BF counts on multiple dates using visual and photographic techniques. The relative abundance of serum proteins in cattle before and after exposure to BFs was analysed using sequential window acquisition of all theoretical fragment ion mass spectrometry (SWATH-MS). Results: Exposure to BFs elicited similar responses in HF and LF cattle, with 79 and 70 proteins, respectively, showing significantly different abundances post exposure as compared to their relevant naïve groups. The comparison of serum samples from naïve HF and LF cattle identified 44 significantly differentially abundant (DA) proteins, while 37 significantly DA proteins were identified from the comparison between HF and LF cattle post-exposure to BFs. Proteins with higher abundance in naïve LF cattle were enriched in blood coagulation mechanisms that were sustained after exposure to BFs. Strong immune response mechanisms were also identified in naïve LF cattle, whereas these responses developed in HF cattle only after exposure to BF. High BF cattle also showed active anticoagulation mechanisms in response to BF exposure, including downregulation of coagulation factor IX and upregulation of antithrombin-III, which might facilitate BF feeding. Conclusion: Underlying differences in the abundance of proteins related to blood coagulation and immune response pathways could potentially provide indirect indicators of susceptibility to BF infestation and biomarkers for selecting more BF-resistant cattle.

Antennal transcriptome analysis reveals sensory receptors potentially associated with host detection in the livestock pest Lucilia cuprina.

BACKGROUND: Lucilia cuprina (Wiedemann, 1830) (Diptera: Calliphoridae) is the main causative agent of flystrike of sheep in Australia and New Zealand. Female flies lay eggs in an open wound or natural orifice, and the developing larvae eat the host's tissues, a condition called myiasis. To improve our understanding of host-seeking behavior, we quantified gene expression in male and female antennae based on their behavior. METHODS: A spatial olfactometer was used to evaluate the olfactory response of L. cuprina mated males and gravid females to fresh or rotting beef. Antennal RNA-Seq analysis was used to identify sensory receptors differentially expressed between groups. RESULTS: Lucilia cuprina females were more attracted to rotten compared to fresh beef (> fivefold increase). However, males and some females did not respond to either type of beef. RNA-Seq analysis was performed on antennae dissected from attracted females, non-attracted females and males. Transcripts encoding sensory receptors from 11 gene families were identified above a threshold (≥ 5 transcript per million) including 49 ATP-binding cassette transporters (ABCs), two ammonium transporters (AMTs), 37 odorant receptors (ORs), 16 ionotropic receptors (IRs), 5 gustatory receptors (GRs), 22 odorant-binding proteins (OBPs), 9 CD36-sensory neuron membrane proteins (CD36/SNMPs), 4 chemosensory proteins (CSPs), 4 myeloid lipid-recognition (ML) and Niemann-Pick C2 disease proteins (ML/NPC2), 2 pickpocket receptors (PPKs) and 3 transient receptor potential channels (TRPs). Differential expression analyses identified sex-biased sensory receptors. CONCLUSIONS: We identified sensory receptors that were differentially expressed between the antennae of both sexes and hence may be associated with host detection by female flies. The most promising for future investigations were as follows: an odorant receptor (LcupOR46) which is female-biased in L. cuprina and Cochliomyia hominivorax Coquerel, 1858; an ABC transporter (ABC G23.1) that was the sole sensory receptor upregulated in the antennae of females attracted to rotting beef compared to non-attracted females; a female-biased ammonia transporter (AMT_Rh50), which was previously associated with ammonium detection in Drosophila melanogaster Meigen, 1830. This is the first report suggesting a possible role for ABC transporters in L. cuprina olfaction and potentially in other insects.

Efficacy of Afoxolaner (NexGard®) in the treatment of furuncular myiasis caused by Dermatobia hominis fly (Diptera: Cuterebridae) in naturally infested dogs.

Furuncular myiasis due to Dermatobia hominis is the second most common skin diseases in dogs that live in tropical climates in Central and South America, causing discomfort and injuring in the connective tissue of the affected dog. Therefore, the objective of the study was to evaluate the effectiveness of Afoxolaner (Nexgard®) in the treatment of canine furuncular myiasis. Twenty-five dogs naturally infested with D. hominis were selected and received a single oral dose of 2.5 mg/kg body weight of Afoxolaner (NexGard®). Larval infestations were classified as light (< 2 larvae), moderate (2 to 5 larvae) and severe (> 5 larvae), according to the number of larvae found in the wound. Twenty-four hours post-treatment, infested lesions were inspected, and all larvae were mechanically removed from the lesion site. All removed larvae were identified as D. hominis larvae and were found dead within 24 h after treatment, demonstrating 100% larvicidal efficacy of Afoxolaner against D. hominis larvae.

Serodiagnosis of nasal myasis in camels (Camelus dromedaries) in Egypt using third larval instar affinity-purified glycoprotein.

The larvae of Cephalopina titillator cause nasopharyngeal myiasis in camels, which parasitize the living tissues of the nasal and paranasal sinuses, pharynx, and larynx. C. titillator infestation adversely affects camel health, meat, and milk production, and can even cause death. In our study, to improve the immunodiagnosis of camel nasal myiasis, a sensitive and specific enzyme-linked immunosorbent assay (ELISA) was developed and evaluated using the Concanavalin-A (Con-A) affinity purification for the C. titillator-N-acetylglucosamine (Ct-GlucNAc) glycoprotein fraction from third larval instars as an antigen for detecting C. titillator antibodies. Crude antigens were prepared from larval instars of C. titillator and evaluated by indirect ELISA. The third C. titillator larval antigen (L3Ct) had the highest protein content (P < 0.001) and the best diagnostic value; chi-square = 235 (P < 0.001). Four glycoprotein fractions were purified separately from the L3Ct antigen by Con-A purification and evaluated. The Ct-GlucNAc glycoprotein fraction was the fraction of choice with the highest diagnostic accuracy (P < 0.05). Using Ct-GlucNAc as a coating antigen, indirect ELISA showed a 99.3% sensitivity for positive results in camel myiasis samples and 100% specificity for negative results in healthy camel samples. The diagnostic accuracy was 99.7%, and no cross reactivity was detected for other parasitic diseases. The indirect ELISA results were confirmed by the western immunoblotting which was characterized by comparing sera from naturally infested dromedary camels with C. titillator, sera from healthy camels and sera from camels with other parasitic infections (Echinococcus granulosus, Fasciola gigantica, Hard ticks; Hyalomma dromedarii, Trichostronglid sp., Eimeria spp., and Cryptosporidium sp.). Immunoreactive antigenic bands of 63, 50, 30 and 18 kDa were predominantly detected in sera from camels with nasopharyngeal myiasis and didn't react with healthy and camel's sera from other parasitic infections. However, seven immunoreactive bands appeared at 120, 70, 63, 48, 35, 29, and 19 kDa in the crude L3Ct antigen. In addition, a positive rate of C. titillator immunodiagnosis was detected by indirect ELISA (48.6%, chi-square = 483, P < 0.001), which was significantly greater than that of postmortem diagnosis (31%). In conclusion, the current study introduces a new diagnostic immunoaffinity glycoprotein fraction of C. titillator 3rd larval instar-based ELISA as a highly accurate, simple and fast method to detect specific antibodies of nasal myiasis in camels.

First Molecular Diagnosis of Oestrus ovis (Linnaeus, 1758) Larvae Causing Conjunctival Ophthalmomyiasis in Mexico City, Mexico.

PURPOSE: Human ophthalmomyiasis is a rare ocular parasitosis that results from the accidental infestation of dipteran larvae of several species, including Oestrus ovis (Linnaeus, 1758). This study aims to present the fourth documented human case of ophthalmomyiasis in Mexico, identifying the etiological agent through morphological and molecular analyses. Additionally, we investigated the phylogenetic position and genetic distances among different specimens globally characterized based on mitochondrial Cox1 sequences. METHODS: A total of five larval specimens were extracted from the patient's eye, with two specimens allocated for identification based on morphological features using a stereomicroscope, and the remaining three preserved in absolute ethanol, one of them used for subsequent analysis using molecular methods. The mitochondrial Cox1 region was amplified and sequenced using automated Sanger sequencing. The resulting sequence was deposited in GenBank under accession number OR440699 and subjected to BlastN analysis against 35 other Cox1 sequences of O. ovis from GenBank. The identity and phylogenetic position of the strains were further explored using parsimony and maximum likelihood phylogenetic methods. RESULTS: Morphological examination of the larval specimens extracted from the patient's eye unequivocally identified them as O. ovis species. BlastN analysis and comprehensive phylogenetic investigations involving a total of 36 Cox1 sequences confirmed the taxonomic identity of the larvae. Notably, our sequence was positioned within the cluster formed by the Brazilian and two Iranian samples. This finding underscores a shared genetic ancestry among these distinct geographical isolates and provides valuable insights into the evolutionary relationships within O. ovis populations. CONCLUSION: The presence of O. ovis infestation in Mexico City suggests potential shifts in environmental conditions favoring fly proliferation, highlighting the need for vigilance in urban healthcare settings.

Molecular epidemiology of the Old World screwworm fly (OWSF) in Iraq, and the genetic structure of various OWSF populations worldwide.

Despite being endemic in Iraq, no reports have been published in the past 10 years to update the molecular epidemiology of the Old World screwworm fly (OWSF), Chrysomya bezziana, in this country. In the present study, 130 sheep from 10 Iraqi governorates were found infected with C. bezziana larvae, whose identities were PCR-confirmed based on the cytochrome b (Cytb) gene, and 23 isolates from various tested governorates were successfully sequenced. Although most isolates (n = 20) belonged to the common haplotype circulating in Iraq, two new haplotypes were detected. Significant changes in OWSF epidemiology in Iraq were also suggested, since infestations were detected, for the first time, in Nineveh governorate. Isolates of the present study were combined to those previously published from Iraq and worldwide, collected after searching the GenBank, and various genetic and population structure analyses were conducted. These isolates displayed a great statistically significant value when tested for the purifying (negative) selection, suggesting the limited occurrence of genetic variations, which was evidenced by the high sequence conservation (C = 0.937) value detected. A few isolates from Africa were revealed during our search, and clustered in a separate lineage other than that of the Asian isolates. The latter displayed different genetic variation patterns when compared. For example, isolates from geographically separate regions, e.g., the Gulf Arab countries and South-Eastern Asia had marked genetic differences. On the other hand, isolates from regions with close geographic proximity (the Gulf Arab countries and Iran) had limited genetic subdivision. This is not the case when comparing isolates from 10 islands in the Indonesian Archipelago. Populations from Sumatra and Sumba were isolated and displayed high genetic variations toward the other populations. On the contrary, populations from Sulawesi, Lombok and Sumbawa displayed limited genetic variations. This is particularly important, since it can help detecting the dynamics of establishing the sterile insect technique over various regions as an effective control strategy against the OWSFs.

Case Report: Ophthalmomyiasis Externa Caused by Cordylobia anthropophaga Masquerading as a Lid Abscess.

Myiasis is a neglected tropical disease caused by the larvae of dipterous flies. Cutaneous infestation is the predominant type documented in sub-Saharan Africa, and ocular involvement is uncommon. We report the rare occurrence of a case of eyelid ophthalmomyiasis caused by Cordylobia anthropophaga in a Nigerian female to raise awareness among practitioners in both tropical and nonendemic areas.

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.

First record of nasopharyngeal myiasis caused by Cephalopina titillator (Clark, 1816) in camel (Camelus dromedarius Linnaeus, 1758) in Uzbekistan.

Nasopharyngeal myiasis caused by the camel nasal bot, Cephalopina titillator, is very common in old world camelids and is usually found at necropsy or during meat inspection. Herein we report massive infection with C. titillator in a 9 years old female one-humped camel slaughtered on February 18, 2024 in the village of Kizil Uy, Nukus District, Republic of Karakalpakstan, northwestern Uzbekistan. A total of 69 larvae: 20 first stage larva (28.9%), 31  second stage larva (44.9%), and 18 third stage larva (26.0%) were detected in the nasal passages and pharynx of the camel. Morphological and morphometrical characters of all larval stages are illustrated and detailed in this article. To our knowledge this is the first record of camel nasal bot infestation in Uzbekistan. Future epidemiological studies are needed to shed light on the prevalence, seasonal fluctuation, clinical impact and economic burden of nasopharyngeal myiasis in dromedary camels of the country.

Uterovaginal prolapse complicated by urogenital myiasis.

Urogenital myiasis is a rare entity that is usually manifested in women belonging to lower socioeconomic status residing in tropical or subtropical nations with poor hygiene or intellectual disability. It is caused by fly larvae capable of penetrating body orifices and healthy or necrotic tissue. The larvae penetrate the skin, forming painful, inflammatory nodules that can form fistula to the internal organs or the skin's surface. Serous and haemorrhagic exudation, along with crawling larvae, are encountered. Some complications, such as secondary infection and tetanus, have been documented. The prognosis is generally good, and treatment consists of removing the parasitic larvae and thoroughly cleansing the affected area.We present a case of a postmenopausal woman with parity 3 and live issue 1 in her early 50s from lower socioeconomic status diagnosed with uterovaginal prolapse complicated by myiasis. The patient was managed successfully with systemic antibiotics, manual removal of myiasis and hysterectomy.

[A case of intestinal myiasis in a bedridden elderly patient].

A 75-year-old man with type 2 diabetes and a history of previous empyema surgery was admitted to our hospital due to difficulty moving caused by chronic obstructive pulmonary disease and dehydration. During the first two days of hospitalization, intestinal myiasis was diagnosed after maggots were found in his diapers. After the maggots disappeared, he developed a fever, prompting antibiotic therapy for a suspected secondary infection, resulting in clinical improvement. Despite thorough home cleaning, no flies or maggots were found, and the source of infection and the fly species remained unknown. Recent reports suggest a higher prevalence of myiasis among the elderly, even with overall improvement in hygiene. While myiasis is typically mild, it is a condition that requires consideration in an aging society. Myiasis is a disease that should be considered in the differential diagnosis of the elderly, especially in people who are bedridden or frail.

Complete Mitochondrial Genome for Lucilia cuprina dorsalis (Diptera: Calliphoridae) from the Northern Territory, Australia.

The Australian sheep blowfly, Lucilia cuprina dorsalis, is a major sheep ectoparasite causing subcutaneous myiasis (flystrike), which can lead to reduced livestock productivity and, in severe instances, death of the affected animals. It is also a primary colonizer of carrion, an efficient pollinator, and used in maggot debridement therapy and forensic investigations. In this study, we report the complete mitochondrial (mt) genome of L. c. dorsalis from the Northern Territory (NT), Australia, where sheep are prohibited animals, unlike the rest of Australia. The mt genome is 15,943 bp in length, comprising 13 protein-coding genes (PCGs), two ribosomal RNAs (rRNAs), 22 transfer RNAs (tRNAs), and a non-coding control region. The gene order of the current mt genome is consistent with the previously published L. cuprina mt genomes. Nucleotide composition revealed an AT bias, accounting for 77.5% of total mt genome nucleotides. Phylogenetic analyses of 56 species/taxa of dipterans indicated that L. c. dorsalis and L. sericata are the closest among all sibling species of the genus Lucilia, which helps to explain species evolution within the family Luciliinae. This study provides the first complete mt genome sequence for L. c. dorsalis derived from the NT, Australia to facilitate species identification and the examination of the evolutionary history of these blowflies.

Cutaneous myiasis caused by Dermatobia hominis (Diptera: Oestridae) in a Polish traveller to South America - a case report.

Myiasis, an infestation caused by dipteran larvae, commonly known as maggots, is one of the most common parasitic skin disorders in the tropical regions. Authors report a case of cutaneous myiasis caused by Dermatobia hominis (Diptera: Oestridae) in a Polish traveller returning from a self-organized trip to South America. Species biology, prophylaxis, and medical implications of this tropical parasitosis are discussed.