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Lachryphagous males of Phortica variegata (Fallén, 1823) are gaining increasing attention in Europe, as they act as vectors of the nematode Thelazia callipaeda Railliet & Henry, 1910, causal agent of thelaziosis, an emergent zoonotic disease. Currently, there are no effective control strategies against the vector, and surveillance and monitoring rely on time-consuming and nonselective sampling methods. Our aim was to improve the knowledge about the population dynamics and the chemical ecology of the species. A total of 5,726 P. variegata flies (96.4% males and 3.6% females, mostly gravid) were collected in field experiments during June–September of 2020 in an oak forest in northern Spain. Our results indicate that 1) by means of sweep netting a significantly higher number of captures were found both around the collector´s body and in the air than at ground level; 2) a positive relationship was detected between the abundance of Phortica flies and temperature, with two significant peaks of abundance at 24 and 33°C; 3) the blend of red wine and cider vinegar was the most attractive bait; 4) yellow traps captured fewer flies compared to black and transparent traps; and 5) a significant reduction toward vinegar and wine was detected in presence of the phenolic monoterpenoid carvacrol. In addition, all the males (n = 690) analyzed by both molecular detection and dissection resulted negative for the presence of T. callipaeda larvae. Overall, these findings provide a better understanding of the vector in terms of monitoring and management strategies., Special thanks are given to the manager of the Izki Golf Club for providing the funding to complete part of the study. M.F. is currently funded by European Union Horizon 2020 Marie Sklodowska-Curie Actions (844285; ‘EpiEcoMod’). We also thank the anonymous reviewers for their many insightful comments and suggestions which helped us to significantly improve the manuscript. This study is part of a work awarded in 2021 with the "III Premio Nacional Felix Pérez y Pérez" of the Illustrious Official College of Veterinarians of Burgos, Spain.

Background Equine thelaziosis is a neglected vector-borne parasitic disease in modern veterinary medicine, lacking recent reports. It is transmitted by Musca autumnalis, and potentially other Muscidae species, by ingesting the lachrymal secretions of its equine host. The distribution of both Thelazia lacrymalis and its intermediate hosts remains largely unknown throughout Europe, with most studies dating back 20 years. The aim of this study was to assess the presence, prevalence and distribution of T. lacrymalis in horses from Romania. Methods The eyes of 273 horses, slaughtered at two abattoirs from the Northwestern and Western regions of Romania, were examined for the presence of T. lacrymalis between March and November 2021. Upon detection, the nematodes were collected and morphologically identified using the keys from literature. Following identification, one specimen from each animal was selected for molecular analysis while the rest underwent detailed morphometric measurements. Mapping and distribution, according to ecoregions, was done using the QGis 3.20 software, while sequences obtained were compared to those available in GenBank through BLAST analysis using the MEGA X software. Results Of the 273 animals sampled, 12 (4.39%) were positive for Thelazia spp. infection. Eighty-seven nematodes were recovered, all morphologically identified as T. lacrymalis. The intensity of infestation varied between one and 33 nematodes/animal while five animals presented a bilateral infestation and seven a unilateral one. The highest prevalence was encountered in Pannonian ecoregion (12.12%) while the lowest was in the Alpine ecoregion (0%). Seventy-five intact specimens underwent detailed morphometric analysis, of the 18–20 parameters, resulting in notable differences in striation lengths compared to the data available in other reports. BLAST analysis identified a 96.46–98.60% similarity to the only other COI gene sequence available for T. lacrymalis. Conclusions The current study represents the first report of T. lacrymalis in horses in Romania. The low prevalence rates are probably linked to the wide use of macrocyclic lactones. Graphic Abstract

For a long time known as the oriental eyeworm, Thelazia callipaeda is a zoonotic nematode that infects the eyes of a wide range of vertebrate hosts including dogs, cats, wildlife carnivores, lagomorphs, and humans. The high occurrence of this infection in Europe and the first cases in the United States have increased scientific interest in the parasite, as it also represents a risk for people living in endemic areas. Therefore, treatment and prevention of thelaziosis in canine population are advocated to reduce the risk of human infection as well. Here, we assessed the efficacy of a formulation containing sarolaner/moxidectin/pyrantel (Simparica TrioThe study was conducted in eyeworm endemic areas of Italy and France, where dogs (n = 125) were assigned into two groups consisting of a negative control group (G1; n = 62), in which animals were treated monthly with a control product (sarolaner; SimparicaIn G1, 16 out of 58 animals (27.6%) were observed with eyeworms during the study, and none of the animals from G2 were ever observed with eyeworms, resulting in 100% efficacy (P 0.0001) in the prevention of establishment of T. callipaeda infection. Adult nematodes and fourth-instar (L4)-stage larvae were recovered from the eyes of positive animals, counted, and morphologically identified as T. callipaeda. In addition, specimens from Italy were molecularly confirmed as belonging to the haplotype 1 (i.e., the only one circulating in Europe so far).Data presented herein demonstrated 100% efficacy of Simparica Trio for the prevention of T. callipaeda eyeworm infection in dogs from highly endemic areas of France and Italy. The use of this formulation is advantageous, as it is a licensed product in Europe with a wide efficacy spectrum against other nematodes, multiple tick species, and fleas. In addition, preventing the development of infection in dogs could also be a prophylaxis measure for zoonotic T. callipaeda infection in humans inhabiting endemic areas.

Background Some species of drosophilid flies belonging to the genus Phortica feed on ocular secretions of mammals, acting as biological vectors of the zoonotic eyeworm Thelazia callipaeda. This study describes an effective breeding protocol of Phortica variegata and Phortica oldenbergi in insectary conditions. Methods Alive gravid flies of P. oldenbergi, P. variegata and Phortica semivirgo were field collected in wooded areas of Lazio region (Italy) and allowed to oviposit singularly to obtain isofamilies. Flies were maintained in ovipots (200 ml) with a plaster-covered bottom to maintain high humidity level inside. Adult feeding was guaranteed by fresh apples and a liquid dietary supplement containing sodium chloride and mucin proteins, while larval development was obtained by Drosophila-like agar feeding medium. The breeding performances of two media were compared: a standard one based on cornmeal flour and an enriched medium based on chestnut flour. All conditions were kept in a climatic chamber with a photoperiod of 14:10 h light:dark, 26 ± 2 °C and 80 ± 10% RH. Results From a total of 130 field-collected Phortica spp., three generations (i.e. F1 = 783, F2 = 109, F3 = 6) were obtained. Phortica oldenbergi was the species with highest breeding performance, being the only species reaching F3. Chestnut-based feeding medium allowed higher adult production and survival probability in both P. oldenbergi and P. variegata. Adult production/female was promising in both species (P. oldenbergi: 13.5 F1/f; P. variegata: 4.5 F1/f). Conclusions This standardized breeding protocol, based on controlled climatic parameters and fly densities, together with the introduction of an enriched chestnut-based feeding medium, allowed to investigate aspects of life history traits of Phortica spp. involved in the transmission of T. callipaeda. Obtaining F3 generation of these species for the first time paved the road for the establishment of stable colonies, an essential requirement for future studies on these vectors in controlled conditions. Graphical abstract

BackgroundThelazia callipaedais a zoonotic parasitic nematode of the family Thelaziidae, withPhortica okadaias its intermediate host and only confirmed vector in China. China has the largest number of human cases of thelaziosis in the world. It is generally believed that infected domestic animals (dogs and cats) are the most important reservoir hosts ofT. callipaeda, and thus pose a direct threat to humans. At present, there is little research or attention focused on the role of wildlife in the transmission cycle of thelaziosis in nature reserves.MethodsWe selected locations in four national nature reserves across China to monitorP. okadaiand wildlife. We used a fly-trap method to monitorP. okadaidensity. Morphological analysis of the parasites collected from the conjunctival sac of the infected wildlife was undertaken as the first step in species identification, and polymerase chain reaction (PCR) was used for species confirmation.ResultsIn 2019, the density ofP. okadaiin Foping National Nature Reserve in China increased sharply, and infectedP. okadaiwere newly found in the reserve. Giant panda, wild boar, leopard cat, and black bear were found to be newly infected withT. callipaeda(one individual of each species). A total of four worms were collected, one from each species of wildlife. The four worms were identified asT. callipaedaby their morphological characteristics; species identification was confirmed by PCR amplification.ConclusionsTo the best of our knowledge, this is the first report ofT. callipaedainfection inP. okadaias well as in a variety of wildlife, including giant panda, in nature reserves in China. These results indicate that there is a transmission cycle ofT. callipaedaamong wildlife in these nature reserves. The increasing number of case reports of thelaziosis in wildlife suggest a likely risk ofT. callipaedainfection for the inhabitants of villages situated around nature reserves.

Introduction Parasitic infections of the eye are a major cause of ocular-surface diseases globally. While most infections are treatable, parasites can cause varying levels of damage mostly due to late diagnosis or misdiagnosis as a result of doctors’ unfamiliarity with their characteristics of latency and crypsis, as well as lack of awareness by the patients. Case Reports In this study, we present three cases of phthiriasis palpebrarum, thelaziasis, and ophthalmomyiasis, respectively. Two of the cases were treated at the clinic and did not recur. One patient refused treatment and was lost to follow-up. Discussion By evaluating the natural histories, morphology, symptoms, clinical findings, and treatment of these parasitic diseases, we systematically analyzed several distinct and unique parasite characteristics, especially latency and crypsis. Furthermore, we have proposed specific examination techniques and methods as well as prevention and treatment strategies from these specific perspectives, aiming to prompt timely diagnoses and early interventions for these diseases by health care workers and improve the public’s awareness of parasitic infections. Conclusion Parasitosis on the ocular surface is a global infectious disease, and prevention strategies include maintaining personal and environmental hygiene and limiting contact with animals. We recommend that health care workers should enhance their ability to detect and diagnose these diseases while promoting the public’s awareness of them in the context of our new perspectives.

Purpose The nematodes of the genus Thelazia are the cause of eye diseases of wild and domestic ruminants throughout the world. The aim of the study was to describe clinical cases of thelasiosis in European bison (Bison bonasus) in Poland, and provide morphometrical features of Thelazia gulosa Railiet and Henry, 1910 and Thelazia skrjabini Erschov, 1928 regarded as potentially useful for species differentiation Methods The conjunctival sacs, tear ducts, the surface of the cornea and nicitating membrane collected from bison were rinsed with saline solution. Any nematodes isolated from the sediment were subjected to morphometric analysis. Results Thirteen of the 16 examined European bison were infected with Thelazia nematodes, belonging to the species T. gulosa and T. skrjabini. The intensity of infection ranged from one to six (mean intensity 5), and four to 29 (mean intensity 14) nematodes T. skrjabini and T. gulosa respectively. Congestion of conjunctival sac, keratitis and corneal opacity, corneal ulceration and perforation as well as purulent eyeball inflammation were observed in infected animals. Conclusions Thelazia gulosa and T. skrjabini can be identified by morphometrical features. As thelasiosis might be a serious threat for protected population of European bison, further studies are needed of the epidemiology and pathology of this emerging parasitosis in Poland.

Ocular infections with Thelazia callipaeda eyeworms in Europe have become more common. We report a case in Hungary caused by T. callipaeda eyeworms in a 45-year-old woman who had no travel history abroad.

Background In August 2020, we found one case of thelazia callipaeda infection during phacoemulsification cataract surgery. This maybe the first report for thelazia callipaeda discovered during phacoemulsification cataract surgery in China. Case presentation An 85 years old farmer was found thelazia callipaeda infection during phacoemulsification cataract surgery. The patient admitted whose foreign body sensation was often found in the right eye in recent 2 months. The worm was then taken out with ophthalmic forceps and put into sterile normal saline. The worm was sent to the Eye Institute of Shandong University of Traditional Chinese Medicine for identification. After identification, the worm was regarded as the male thelazia callipaeda. The head is blunt and round, the tail end curls to the abdomen, and the long copulation spines protrudes from the cloaca. The conjunctival sac was washed carefully with a large amount of Sodium Lactate Ringer ‘s Solution. After operation, antibiotics, pranoprofen eye drops, and tobramycin dexamethasone eye drops were further applied. After continuous examination of conjunctival sac for 2 weeks, the patient’s visual acuity maintained 20/20, confirming that there was no residual thelazia. Conclusions This report highlights the physician should ask the patient’s history carefully before operation and it is necessary to strengthen health publicity and education, maintaining clean environment and keeping personal eye hygiene.

Thelaziasis is a zoonotic disease which affects the eye of domestic and wild carnivores caused by the nematode Thelazia. It is transmitted to humans by secretophagous arthropod-borne zoophilic nonbiting flies of the family Drosophilidae. Human thelaziasis is rare and occurs in poor socio-economic families of the rural locations where people live in close proximity with animals. A one and halfyear-old was presented to the outpatient ophthalmology clinic after her mother noticed a whitish, thread-like worm in her right eye. A total of four worms were mechanically removed from her right eye. All the collected worms were gravid female nematodes of Thelazia species. The present case of human ocular thelaziasis from Palpa, Nepal is presented for its rarity.

Background The present clinical field trial was conducted to assess the efficacy of a broad-spectrum parasiticide spot-on formulation containing eprinomectin (Broadline®) against Thelazia callipaeda eyeworm in naturally infected cats. Methods Fifteen privately owned cats harboring at least one live adult T. callipaeda were included in the study. Cats were randomly allocated to an untreated control group of seven cats or to a Broadline®-treated group of eight cats. Cats were treated on Day 0; ocular examinations were performed at inclusion and on Days 7 and 14; eyeworms were recovered and counted on Day 14. The primary efficacy assessment was based on group comparison of number of T. callipaeda on Day 14. Results Seven days after treatment, six of eight treated cats were negative for eyeworm infection per visual examination, and on Day 14 no eyeworms were found in the treated cats while the seven untreated cats were still infected (geometric mean: 1.97). All cats had inflammatory ocular signs at inclusion; on Day 14, five of eight treated cats had recovered while all untreated control cats were still symptomatic. All collected parasites were confirmed to be T. callipaeda by morphology and molecular characterization. Conclusions A single treatment with Broadline® provided 100% efficacy against feline thelaziosis and improved related ocular inflammation signs. Graphical abstract

Spirocerca lupi is the etiological agent of spirocercosis in canids from tropics and subtropics in the world. This parasite can lead to life threatening complications, including the formation of fibro and osteosarcomas. A 5-year- old crossbreed Maltese dog from Costa Rica was presented to a veterinary clinic with two nodules in the sub- cutaneous tissues of the cervical region. One of the nodules resolved with intramuscular antibiotic and anti- inflammatory treatment. However, the other nodule persisted and became purulent. The content of the nodule was excised releasing two coiled 3-cm reddish worms. Identity of the specimens was confirmed by amplifying an 850 bp fragment of the 18S rDNA. The obtained sequence showed 99.96% similarities with S. lupi from Hungary. The present study highlights the need of veterinary clinicians' awareness towards nematode aberrant migrations, and the require of molecular studies to gain insight into the parasite's evolutionary history. Spirocerca lupi es el agente etiológico de la espirocerosis en los cánidos de los trópicos y subtrópicos del mundo. Este parásito puede provocar complicaciones mortales, como la formación de fibrosis y osteosarcomas. Un perro maltés de 5 años de edad, procedente de Costa Rica, acudió a una clínica veterinaria con dos nódulos en los tejidos subcutáneos de la región cervical. Uno de los nódulos se resolvió con un tratamiento antibiótico y antiinflamatorio intramuscular. Sin embargo, el otro nódulo persistió y se volvió purulento. El contenido del nódulo fue extirpado liberando dos gusanos rojizos enrollados de 3 cm. La identidad de los especímenes se confirmó mediante amplificación de un fragmento de 850 pb del ADNr 18S. La secuencia obtenida mostró una similitud del 99,96% con S. lupi de Hungría. El presente estudio pone de relieve la necesidad de que los clínicos veterinarios tomen conciencia de las migraciones aberrantes de los nematodos, y de que se requiera una mayor concienciación al respecto y la necesidad de realizar estudios moleculares para conocer la historia evolutiva del parásito. Universidad Nacional, Costa Rica Escuela de Medicina Veterinaria

Background The present clinical field trial was conducted to assess the efficacy of a broad-spectrum parasiticide spot-on formulation containing eprinomectin (Broadline®) against Thelazia callipaeda eyeworm in naturally infected cats. Methods Fifteen privately owned cats harboring at least one live adult T. callipaeda were included in the study. Cats were randomly allocated to an untreated control group of seven cats or to a Broadline®-treated group of eight cats. Cats were treated on Day 0; ocular examinations were performed at inclusion and on Days 7 and 14; eyeworms were recovered and counted on Day 14. The primary efficacy assessment was based on group comparison of number of T. callipaeda on Day 14. Results Seven days after treatment, six of eight treated cats were negative for eyeworm infection per visual examination, and on Day 14 no eyeworms were found in the treated cats while the seven untreated cats were still infected (geometric mean: 1.97). All cats had inflammatory ocular signs at inclusion; on Day 14, five of eight treated cats had recovered while all untreated control cats were still symptomatic. All collected parasites were confirmed to be T. callipaeda by morphology and molecular characterization. Conclusions A single treatment with Broadline® provided 100% efficacy against feline thelaziosis and improved related ocular inflammation signs. Graphical abstract

Introduction: We report a local human case of Thelazia callipaeda eye infection in a 49-year-old lady with history of fly contact in Hong Kong. Case description: A 49-year-old lady presented with right eye foreign body sensation for one month. She recalled a fly being stuck onto her right upper eyelashes with mascara when she went hiking in a forest trail in Hong Kong. On assessment there were a lot of giant papillae on palpebral conjunctiva. Three living worms crawling on conjunctiva were discovered and removed in total. The worms were identified as Thelazia callipaeda by morphology and molecular sequencing. After removal, her symptoms resolved completely. Conclusion: Human thelaziasis is probably under-reported in many countries. The presence of giant papillary conjunctivitis in non-contact lens wearers should alert clinicians to the possibility of thelaziasis in patients with compatible exposure history in endemic regions. Ophthalmologists should increase their awareness towards this uncommon disease and should not wrongly attributed the symptoms to allergic conjunctivitis.

The zoonotic parasitic nematode Thelazia callipaeda, also known as the oriental eye worm, is endemic in several European countries, including Portugal. Infections may result in ocular disease in domestic and wild animals as well as humans, with more or less severe manifestations. We report the first human case of ocular thelaziosis by T. callipaeda in Portugal, a country where the parasite had already been found to infect dogs, cats, red foxes, wild rabbits and a beech marten. An 80-year-old patient from east-central Portugal, who had been suffering from tearing for a few years, had whitish filiform fragments removed from the left eye. Polymerase chain reaction of partial cytochrome c oxidase subunit 1 and 18S small subunit rRNA genes followed by bidirectional sequencing and BLAST analysis confirmed T. callipaeda haplotype 1, the only haplotype previously reported in Europe. The endemicity of T. callipaeda in domestic and wild animals in east-central Portugal makes it very likely that infection of the human patient had occurred locally. In east-central and other geographical areas of Portugal, veterinarians and physicians, especially ophthalmologists, should regard T. callipaeda as a cause of ocular pathology in animals and humans.

International audience; Based on light and electron microscopical studies, a new nematode parasite, Echinocephalus inserratus sp. n. (Spirurida: Gnathostomatidae), is described from the spiral valve of the broad cowtail stingray Pastinachus ater (Macleay) (Dasyatidae, Myliobatiformes) from off New Caledonia. The new species is morphologically and biometrically most similar to Echinocephalus overstreeti Deardorff et Ko, 1983, differing from it mainly in the absence of serrations on the posterior parts of pseudolabia and on interlabia, and in having a longer gubernaculum (150-299 µm long). Morphologically unidentifiable, mostly encapsulated larvae of Echinocephalus spp. were recorded from the following six species of teleost fishes collected in New Caledonian waters, serving as paratenic hosts: Perciformes: Acanthopagrus berda (Forsskål) (Sparidae) and Nemipterus furcosus (Valenciennes) (Nemipteridae); Tetraodontiformes: Abalistes stellatus (Anonymous), Pseudobalistes fuscus (Bloch et Schneider) (both Balistidae), Lagocephalus sceleratus (Gmelin) (Tetraodontidae) and Aluterus monoceros (Linnaeus) (Monacanthidae). Co-parasitising larvae of Ascarophis sp. and Hysterothylacium sp. were also collected from P. fuscus. All these findings represent new host and geographical records. A key to valid species of Echinocephalus Molin, 1858 is provided.

We report a case of autochthonous infection of the eye worm Thelazia callipaeda in a dog in the northeastern United States. Integrated morphologic identification and molecular diagnosis confirmed the species. Phylogenetic analysis suggested introduction from Europe. The zoonotic potential of this parasite warrants broader surveillance and increased awareness among physicians and veterinarians.

Thelaziosis caused by the nematodeWe retrospectively analysed the occurrence of feline thelaziosis (study 1), examined its prevalence based on cross-sectional sampling of endemic areas (study 2), and assessed the therapeutic/prophylactic benefits of a spot-on (moxidectin) and therapeutic efficacy of an eye drop (ivermectin) formulation (study 3).In study 1, 69Our data confirm the endemic status of

Spirocerca lupi is the etiologic agent of spirocercosis in canids from the tropical and subtropical regions of the world. A 1-year-old dog was presented with weight loss and diarrhea. Upon examination, thickened limbs, a mass in the lung’s right middle lobe and an esophageal mass were found. The pulmonary mass aspirate revealed S. lupi eggs surrounded by a purulent infiltrate. Limb bone biopsies showed irregular mineralization and cartilaginous metaplasia. Treatment with doramectin resolved the initial clinical signs. However, regurgitation was reported 5 months later. Therefore, the dog received additional doramectin and later vomited a worm morphologically consistent with a S. lupi adult. Herein we report the esophageal migration of S. lupi with egg deposition to the lung parenchyma, accompanied by secondary hypertrophic osteopathy. This report highlights the presence of S. lupi in Costa Rica and the importance of including this parasite as a differential diagnosis when compatible signs are identified. Spirocerca lupi es el agente etiológico de la espirocercosis en los cánidos de las regiones tropicales y subtropicales del mundo. Un perro de 1 año se presentó con pérdida de peso y diarrea. En la exploración se encontraron extremidades engrosadas, una masa en el lóbulo medio del pulmón derecho y una masa esofágica. El aspirado de la masa pulmonar reveló huevos de S. lupi rodeados de un infiltrado purulento. Las biopsias óseas de las extremidades mostraron una mineralización irregular y metaplasia cartilaginosa. El tratamiento con doramectina resolvió los signos clínicos iniciales. Sin embargo, 5 meses más tarde se observó una regurgitación. Por lo tanto, el perro recibió doramectina adicional y posteriormente vomitó un gusano morfológicamente consistente con un adulto de S. lupi. Aquí informamos de la migración esofágica de S. lupi con deposición de huevos al parénquima pulmonar, acompañada de una osteopatía hipertrófica secundaria. Este informe destaca la presencia de S. lupi en Costa Rica y la importancia de incluir este parásito como diagnóstico diferencial cuando se identifican signos compatibles. Clínica Veterinaria Best for Pets, Costa Rica. Laboratorio Clínico Veterinario, Costa Rica. Escuela de Medicina y Cirugía Veterinaria San Francisco de Asis, Costa Rica. Universidad Nacional, Costa Rica. Escuela de Medicina Veterinaria

Because of their free-ranging nature, the probability of wild animals being exposed to vector-borne pathogens is likely higher than that of humans and pets. Recently, the red fox (Vulpes vulpes) has been suspected as being a reservoir or host of several pathogens of veterinary and public health importance. We conducted a molecular survey on 93 red foxes hunted in 2008-18, in the departments of Bouches-du-Rhone and Var, in southeastern France, for pathogens including Leishmania infantum, Piroplasmida, Hepatozoon spp., nematodes, Coxiella burnetii, Borrelia spp., Rickettsia spp., and Anaplasmataceae. Spleen samples were screened for the presence of vector-borne pathogens by PCR followed by sequencing. Pathogens were detected in 94% (87/93) of red foxes, and coinfections were identified in 24% (22/93) of foxes. We identified DNA from Hepatozoon canis, L. infantum, and Babesia vogeli in 92% (86/93), 15% (14/93), and 3% (3/93) of red foxes, respectively. We also found DNA of nematodes in 3% (3/93) of foxes; Spirocerca vulpis was identified in one fox and Dirofilaria immitis in the two others. Interestingly, C. burnetii genotype 3, previously described in humans from the same region, was identified in 3% (3/93) of foxes and Anaplasma platys in 2% (2/93) of foxes. We did not detect DNA of Borrelia spp., Bartonella spp., or Rickettsia spp. In our study, the prevalence of pathogens did not vary by fox origin, sex, or tick carriage. Molecular evidence of B. vogeli, H. canis, S. vulpis, D. immitis, C. burnetii, and A. platys in red foxes has not previously, to our knowledge, been reported from southern France. We propose that red foxes are potential reservoirs for several pathogens, including major zoonotic agents such as L. infantum. They could be incidental hosts for pathogens, such C. burnetii. The high prevalence for H. canis suggests an important role of foxes in domestic dog (Canis lupus familiaris) infection. These animals may pose a threat to human and animal health.

Spirocerca lupi infection in dogs (Canis domesticus) is associated with esophageal lesions that may evolve to a neoplastic stage in the form of esophageal sarcoma. In the red fox (Vulpes vulpes) infected with the closely related Spirocerca vulpis, similar lesions may occur in the stomach, but neoplastic forms have not been reported. We characterize Spirocerca vulpis-induced lesions in the fox, using pathology and immunohistochemical (IHC) techniques. Seventy-one out of 163 Spirocerca vulpis-positive red foxes were selected and subjected to histopathological study. Lesions were classified as patchy or diffuse. Ten patchy and 10 diffuse lesion samples were studied using three IHC markers (CD68, CD3, and CD79α for macrophages, T lymphocytes, and B lymphocytes, respectively) and H&E stain for neutrophils and eosinophils. Intensity of necrosis, hemorrhages, and the presence of collagen was also analyzed. Of the S. vulpis-positive red foxes, 96.9% had S. vulpis nodules localized in the gastric area (wall and/or omentum), and 3.1% had nodules in the small intestine. All the samples had a moderate to severe lymphoplasmacytic infiltrate. Mild eosinophil infiltration was observed in both types of lesions, while neutrophil infiltration was significatively higher in the patchy than in the diffuse lesions. Fibrosis with mature collagen fibers was also predominant in the patchy lesions along with the presence of T lymphocytes and macrophages. Both the patchy and diffuse patterns had very few B lymphocytes. These findings suggest that the diffuse form is an earlier stage of the lesion, which eventually evolves into patchy forms. Neoplastic forms were not seen. Although more studies are necessary, this study describes the lesions, characterizes the inflammatory infiltrates, and establishes a possible evolution of the different pathological forms of S. vulpis infection in the red fox.

The eye nematode Thelazia callipaeda Railliet et Henry, 1910 (Spirurida: Thelaziidae) is a vector-borne zoonotic nematode infecting a range of wild and domestic carnivores as well as humans. It is considered to be a causative agent of emerging and neglected disease and currently invades central part of Europe. Nematodes were collected from the eye of a dog living in Prague, which never travelled outside the Czech Republic. The nematodes were identified based on their morphology and partial sequence of the cox1 gene as T. callipaeda haplotype 1. This finding represents the northernmost record of autochthonous canine thelaziosis in Europe. The insufficient control of imported animals as well as free movement of dogs and wild carnivores within Europe probably facilitates spreading of T. callipaeda throughout the continent. To better understand the spreading of T. callipaeda and to prevent its zoonotic transmissions, information about the risk of this infection in newly invaded countries should be disseminated not only among veterinarians and physicians, but also within the community of pet owners and hunters.

The first case of feline ocular Thelazia callipaeda infection and two new canine imported infections in West Germany are here described. The three animals had a history of recent travel to/from other countries. The young adult cat imported from Spain presented an intermittent unilateral ocular discharge. During in-depth ophthalmic examination, a single alive nematode was removed from the conjunctival compartment of the affected eye. Referring to the canine cases, an adult female dog originated from Kenya presented epiphora and mucous whitish-grey discharge of the right eye. During flushing of the nasolacrimal duct two small, thin and long nematodes were removed. Furthermore, a male Borzoi racing dog with regular visit to racing tracks in different countries presented ocular mucous discharge. At ophthalmologic examination, two transparent-whitish vital nematodes were removed. All nematode specimens of the three cases were morphologically identified as adult T. callipaeda parasites. The animals were treated orally with milbemycin oxime (2.0 mg/kg; cat) or milbemycin oxime/praziquantel (0.5 mg/kg and 5.0 mg/kg; dogs) twice with 1-week interval resulting in complete resolution of symptoms. The repeated introduction of patent T. callipaeda-infected animals, especially from southern and eastern endemic countries, will ease the establishment of ophthalmic thelaziosis in Northern Europe. The male fruit fly, Phortica variegata, an intermediate host of T. callipaeda, is endemic within European countries. Considering the clinical and zoonotic relevance of ophthalmic thelaziosis, enhanced disease awareness of European medical and veterinarian doctors and in-depth eye examination for proper detection of T. callipaeda are crucial for appropriate anthelmintic treatments and to limit spreading of the infection.

A 41-year-old man from Son La Province, Vietnam, sought care for disseminated prurigo-like skin lesions from which nematode larvae were emerging. We morphologically and molecularly identified the larvae as Oxyspirura sp. Our findings confirm this nematode species as a zoonotic pathogen for emerging disease.

A 12-year old Elo dog was presented with recurring symptoms of conjunctivitis in November 2019. A single whitish nematode was found upon inspection of the eye and identified as a Thelazia callipaeda male. The morphological identification of the eye worm was supported by analysis of a partial cytochrome c oxidase I (cox1) gene sequence. The dog lived in Lower Saxony, northwestern Germany, and had not visited regions known to be endemic for T. callipaeda. This suggests that a local transmission cycle of this zoonotic nematode may exist in Germany.

BackgroundThe drosophilidPhortica variegatais known as vector ofThelazia callipaeda, the oriental eyeworm native to Asia that has become an emergent zoonotic agent in several European regions. Unlike almost all other arthropod vectors of pathogens, onlyP. variegatamales feed of lachrymal secretions of animals, ingesting first-stage larvae (L1) of the worm living in the orbital cavities of the host, and allowing with the same behaviour the introduction of infective L3. Despite the increased detection ofT. callipaedain many European countries, information about the length of the lachryphagous activity period ofP. variegataand a deep knowledge of the environmental and climatic variables involved are still limited.MethodsWe herein present the results of a multicentre study involving five sites from four different countries (Italy, Spain, UK and USA) where canine thelaziosis is endemic and/or where it has already been ascertained the presence ofP. variegata. Field data have been obtained on a fortnightly basis from mid-April to the end of November 2018 from a contemporary standardized sampling (same sampling effort and time of collection in all sites) of lachryphagous flies collected around the eyes of a human bait using an entomological net. These data have been associated to data collection of local climatic variables (day length, temperature, wind speed, barometric pressure and relative humidity).ResultsOverall, a total of 4862P. variegataflies (4637 males and 224 females) were collected, with high differences in densities among the different sampling sites. Significant positive correlations were found betweenP. variegatamale density and temperature and wind speed, while negative correlations were observed for barometric pressure and relative humidity. However, the above significant differences are confirmed in each sampling site separately only for the temperature.ConclusionsThis multicentre study highlights that temperature is the major common environmental driver in describing the lachryphagous activity ofP. variegatain Europe and USA and, therefore, the transmission risk of thelaziosis.

Thelazia callipaeda (Spirurida, Thelaziidae) is a vector-borne zoonotic nematode parasite of the conjunctival sac of a large spectrum of hosts, currently having an expanding distribution range throughout Europe. In Romania, the parasite has a relatively large distribution range and host spectrum. However, the presence of T. callipaeda in red foxes ( Vulpes vulpes) has not yet been confirmed. The aim of the present study was to assess the prevalence and geographical distribution of T. callipaeda in red foxes from Romania. A total of 514 animals originating from 10 counties of western Romania were evaluated for the presence of T. callipaeda. Overall, 151 red foxes (29.38%) were positive. No significant differences were noticed between sex and age groups of positive foxes. In total, 3,350 adult nematodes were recovered. The intensity of infection had a mean value of 23.2, while the sex ratio was 2.2:1 in favor of females. In 91 foxes, both eyes were affected, while 60 harbored unilateral infection. Sequence analysis of the cox 1 gene of one nematode from each positive fox ( n=151) revealed the exclusive presence of haplotype h1. We report a relatively high prevalence and distribution range of T. callipaeda infection in red foxes in Romania, highlighting their potential role as a wildlife reservoir.

Background In Europe, the first Thelazia callipaeda infections were found in the eyes of some dogs in Italy three decades ago. Since that time, this vector-borne nematode species has been diagnosed in domestic and wild carnivores and humans in some western European countries. During the last few years, autochthonous thelaziosis of dogs, red foxes, cats and humans has also been reported from eastern Europe. The first cases of ocular infections caused by T. callipaeda have been described in dogs living in the eastern and southern part of Slovakia and Hungary. Methods Whitish parasites found in the conjuctival sac and/or under the third eyelid of one or both eyes of animals were removed and morphologically identified according to species and sex. To confirm the morphological identification with molecular analysis a single step conventional PCR was carried out. Results A total of 116 adult worms (1–37 per dog, median: 7, IQR: 14.5 and 7 from a cat) were collected from the eyes of 11 animals. Nematodes were identified as T. callipaeda according to the morphological keys and molecular analysis. The sequences of a portion of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene were identical to those representing T. callipaeda haplotype 1, previously reported in neighbouring and other European countries. Since the infected cat and dogs had never travelled abroad, all of the cases were autochthonous thelaziosis. Conclusions The present study reports the first case of thelaziosis in a cat and new cases in 10 dogs found in the southern and northern region of Hungary, respectively. Further studies are needed to clarify whether wild carnivores (e.g. red foxes, golden jackals) may act as reservoirs of this eyeworm species in the country.

Thelaziasis is an ocular infection caused by nematodes of Thelazia spp., usually transmitted by flies that feed on lacrimal secretions. Although a common veterinary infection, human infection is considered to be a rare zoonotic event. Cases of human thelaziasis have been reported worldwide, predominantly in Europe and Asia.1 Cases occur predominantly in rural communities with close proximity to animals and poor living standards. Thelaziasis mainly affects the elderly and children, who may be less able to keep flies away from their faces.2 Most of the reported cases have been caused by Thelazia callipaeda, occurring in the old world, particularly in Japan.1 Thelazia callipaeda has also rarely been reported infecting humans in China, Japan, India, Burma, Korea, Taiwan, Thailand, Indonesia, Russia, Italy, and France.3 There have been 10 previously reported cases of human thelaziasis from the United States; nine from California and one in Utah, all were reported as Thelazia californiensis.4,5 Other species of Thelazia are reported as infecting animals in the United States, these being, Thelazia gulosa, Thelazia lacrymalis, Thelazia rhodesi, and Thelazia skrjabini.3

Background The fruit fly Phortica variegata (Drosophilidae: Steganinae) feeds on the ocular secretions of animals and humans, and has been described as an intermediate host of the eye worm Thelazia callipaeda (Spirurida: Thelaziidae) in Italy. Despite the increased detection of T. callipaeda in many European countries, information about its vector role in natural conditions is still limited. In the Iberian Peninsula, thelaziosis caused by T. callipaeda has been reported in dogs, cats, red foxes, wild rabbits and humans. Methods In the last seven years, we have detected increased numbers of cases of canine thelaziosis at three locations in mainland Spain: Site 1, La Vera region (Cáceres Province, central-western Spain; 51 cases); Site 2, El Escorial municipality (Madrid Community, central Spain; 23 cases); and Site 3, Miraflores de la Sierra municipality (Madrid Community, central Spain; 41 cases). Site 1 is considered endemic for T. callipaeda while the other two sites have been recently recognised as risk zones for T. callipaeda infection. Results From June 2016 to September 2017, 2162 flies were collected and morphologically identified as Phortica spp. (Site 1, n = 395; Site 2, n = 1544; and Site 3, n = 223). Upon dissection, third-stage T. callipaeda larvae were found in two out of 155 flies examined from Site 1, and both these larvae tested molecularly positive for the eye worm. Of the 395 flies collected from Site 1, 371 were molecularly processed for arthropod species identification and T. callipaeda detection. All 371 flies were identified as P. variegata and 28 (7.5%; 95% CI: 4.8–10%) tested positive for T. callipaeda DNA haplotype 1. Conclusions Our findings indicate that T. callipaeda circulates among dogs and P. variegata in Spain, where zoonotic cases have been also reported. The co-existence of canine thelaziosis and Phortica spp. in geographical areas previously considered free of the eye worm indicates a risk of infection for both animals and humans living in this region.

Ocular thelaziosis is caused by nematodes of the genus Thelazia (Spirurida, Thelaziidae), which inhabit the surface of the eyes and associated tissues. Thelazia callipaeda affects a range of mammal species, including humans, and in the last two decades has been reported in multiple European countries, being classified as an emergent vector-borne pathogen. In Portugal T. callipaeda is endemic in north-eastern areas, where it has been reported in domestic dogs, cats, red foxes (Vulpes vulpes) and wild European rabbits (Oryctolagus cuniculus). We report, for the first time, T. callipaeda in a beech marten (Martes foina) from Portugal and highlight the presence of haplotype 1 as the only one found in Europe, irrespective of the host species and geographical area of provenience.

Canine spirocercosis is caused by the nematode Spirocerca lupi. Migration results in oesophageal fibro-inflammatory nodules that may undergo neoplastic transformation. No studies have assessed pre- or post-surgical prognostic indicators in dogs that undergo intervention for S. lupi induced oesophageal neoplasia. This observational, multi-center study aimed to assess the outcome of dogs with Spirocerca induced sarcoma undergoing endoscopic-guided ablation (n = 12) or surgery (n = 18), and identify prognostic indicators. Parameters evaluated included: age, weight, gender, presenting complaints, duration of clinical signs, complete blood count, serum biochemistry, neoplasia size, placement of percutaneous endoscopically-placed gastrostomy tube, histopathological mitotic indices, days to discharge and chemotherapy administration. Kaplan-Meier survival curves showed no difference in survival between ablation and surgery {(median: 73.5 days (range: 0–1511) vs. 108 days (range: 0–1550), respectively (p = 0.982)}. Reduced survival was documented in patients presenting with weight loss (P = 0.027), hypochromasia (MCHC 15 × 109/L, P = 0.017) with a hazard ratio of 2.51 (CI95% = 1.071–6.018, P = 0.034), 2.71 (CI95% = 1.10–6.65, P = 0.03) and 4.39 (CI95%: 1.21–15.97, P = 0.025) respectively. In the dogs surviving more than 21 days, Ht 15.0 × 109/L at presentation were associated with reduced survival (p = 0.016, p = 0.021 respectively) and hazard ratio of 3.29 (CI95% = 1.18–9.2, P = 0.023) and 3.81 (CI95% = 1.15–12.55, P = 0.028) respectively. Intra-intervention-group survival analysis identified increased survival time in dogs receiving chemotherapy, but only within the surgical group (P = 0.02).The hospitalisation time of dogs undergoing ablation (median: 0 days, range: 0–4) was significantly shorter than dogs undergoing surgery (9 days, 1–21) (P In this study, no clear benefit was identified for surgery, thus when ablation is technically possible it should be considered advantageous, as hospitalisation time is significantly shorter. Weight loss, hypochromasia and leucocytosis were identified as long-term prognostic indicators at presentation.

Spirocerca lupi is a parasitic nematode that causes spirocercosis predominantly in domestic dogs. Spirocerca lupi nematode samples were collected from four regions around South Africa and analyzed to compare the genetic diversity among the regions. A total of 56 S. lupi nematodes were obtained by necropsy from domestic dogs and wild black-backed jackals (Canis mesomelas). Sixteen different haplotypes of cox1 were identified some of which are shared between regions as well as with black-backed jackal. The genetic similarity between S. lupi in domestic dogs and black-backed jackals indicates transmission between these canid species and may have potential conservation implications.

Background The pathology of spirocercosis, a disease caused by the infestation of carnivores with the nematode Spirocerca lupi, has been extensively described in domestic dogs and coyotes. However, it has not been described in wild carnivores in South Africa. The aim of this study was to evaluate whether black-backed jackals are a host for Spirocerca species and to provide a detailed description of the associated pathology. Jackals were also stratified according to age and the Spirocerca species recovered were characterized using molecular techniques. Methods Standard necropsies were performed on routinely culled jackals from three of the nine provinces of South Africa during the period June 2012 to February 2013. Jackals were screened for the presence of pathognomonic Spirocerca-induced lesions and for evidence of aberrant migration. Relevant samples were submitted for histopathology and collected larvae were genotyped at nine microsatellite loci. Results Spirocerca lupi-associated aortic lesions were found in 16 of 93 (17%) black-backed jackals. Of these, four (25%) were associated with S. lupi larvae. Genotyping of the larvae revealed amplification of all nine loci that amplified dog-derived S. lupi, with the same level of polymorphism in the allele size ranges. Only 1 of 93 jackals had an esophageal nodule with concurrent S. lupi-induced aortic aneurysms. The single esophageal nodule found did not contain adult nematodes, nor did it communicate with the esophageal lumen. None of the jackals that were examined had macroscopically evident spondylitis, which is frequently reported in the dog. Histopathology of the S. lupi-induced aortic lesions in the jackal revealed replacement of elastic and smooth muscle fibers by fibrous connective tissue. In cases where inflammation was present, the inflammatory infiltrate consisted predominantly of eosinophils. The single esophageal nodule histologically resembled the early inflammatory nodule described in dogs and consisted of fibrous connective tissue, multifocal accumulation of lymphocytes, plasma cells and rare hemosiderin-laden macrophages. Conclusions These lesions suggest that the life cycle of S. lupi may not or only rarely be completed in jackals. A possible explanation might be that jackals are relatively resistant to developing significant pathology associated with S. lupi-infection. However, before any conclusions can be drawn, many more jackals, including those that die naturally will have to be investigated for evidence of S. lupi infection.