Social media as a tool for detecting underdiagnosed parasitic infections: the case of spirocercosis.

Spirocercosis is a parasitic disease caused by the nematode Spirocerca lupi that affects several canids around the world. In Costa Rica, few spirocercosis cases have been reported; however, it is suspected that the infection is currently underdiagnosed. As seen during the COVID-19 pandemic, social media has sped up the spread of scientific knowledge, raising practioner's awareness of the epidemiology and signs and manifestations of infections. The aim of this work was to use social media as a tool for increasing awareness of spirocercosis among veterinarians and investigate the frequency of cases occurring in Costa Rica. Infographic material about the infection was disseminated by Whatsapp, Facebook, and Instagram from January 2021 to July 2022 using a public account of the Laboratory of Helminthology of the University of Costa Rica. In addition, Google Forms questionnaires were completed by veterinarians to report the number of spirocercosis cases in their clinics, associated manifestations, diagnosis, treatment, and outcome of the disease. Remarkably, since 2020, a six-fold increase in the number of cases per year was noted. Clinical manifestations matched those reported elsewhere, and one of every three dogs infected with S. lupi died spontaneously or required euthanasia because of the infection. This work suggests that S. lupi infection in Costa Rica may have been underdiagnosed in the last decade, and the knowledge and awareness of clinical personnel play a major role in the detection of this and other infectious diseases.

Diversity of Thelazia spp. in domestic cattle from Romania: epidemiology and molecular diagnosis by a novel multiplex PCR.

BACKGROUND: Thelaziosis is a neglected vector-borne disease caused by parasitic nematode worms of the genus Thelazia which affects various hosts. Limited attention has been given to ungulate-associated Thelazia species. Current diagnosis of thelaziosis and the identification/differentiation of species heavily relies on morphological features. Therefore, we conducted an epidemiological study in Romanian cattle, with the aim to obtain morphological and molecular data that can be used for species identification. METHODS: The eyes of 705 slaughtered cattle were sampled and subjected to morphological identification, morphometric analysis, and molecular characterization. PCR amplification and sequence analysis were performed based on the cytochromec oxidase subunit 1 (COI) gene. Statistical tests assessed the correlations between infection parameters and ecological or biogeographical factors. A novel PCR method was developed based on the consensus sequence from each species. Specific forward primers were designed for each of the three species, and a reverse primer (COIintR) was used for all reactions. A consensus thermal profile was established by gradient PCR amplification of each species separately. RESULTS: Of the sampled cattle, 19.3% were infected with Thelazia spp. Prevalence varied significantly with ecogeographical factors. A total of 585 Thelazia nematodes were recovered, with T. rhodesi being the most abundant, followed by T. skrjabini and T. gulosa. Morphometric and molecular analyses supported the morphological identification, yielding unique sequences for each species. From the 59 T. rhodesi specimens sequenced, 29 unique sequences were obtained, with a 99.1-99.85% nucleotide identity to the only other COI sequence present in GenBank®. All nine T. gulosa isolates were unique (99.37-100% nucleotide identity to other sequences), while T. skrjabini specimens displayed 98.47-100% nucleotide identity to the sole available sequence. CONCLUSIONS: Bovine thelaziosis is prevalent in Romania, raising concerns for animal welfare and potential economic impacts. Infected cattle grazing alongside vulnerable wild ruminants, such as the European bison, may affect conservation efforts. Our newly developed multiplex PCR shows promise as a valuable surveillance tool, enabling the detection of occult infections in apparently healthy animals through lachrymal secretion testing.

Ocular thelaziosis: an emerging human parasitosis in Europe.

Ocular thelaziosis is a parasitosis distributed mainly in East Asia, but increasingly described in Europe in different domestic and wild animals, including dogs, different wild canids and lagomorphs, and exceptionally in humans. In Spain, in some areas, a high infection prevalence rate has been described in domestic canids, which may lead to an increase in human cases. However, the description of human cases is still exceptional, which suggests that they are probably underdiagnosed. A case of ocular thelaziosis in a 2-year-old girl from an urban environment is described.

Spirocerca lupi in the stomach of two Andean foxes (Lycalopex culpaeus) from Chile.

The genus Spirocerca includes nematodes that parasitize the stomach and the oesophagus of carnivores, chiefly canids. Herein, we provide new data about the morphological, histopathological, and molecular characterization of Spirocerca sp. in Andean foxes (Lycalopex culpaeus) in Chile. Intact immature worms, identified as Spirocerca sp., were recovered in the lumen of the stomach from two foxes. Histologically, worms morphologically consistent with spirurid nematodes were present within the wall of the stomach and surrounded by nodular areas of inflammation with central necrotic debris. Molecular analysis of the cox1 gene yielded 19 sequences and 5 nucleotide sequence types with 99.95 to 99.98% similarity, being shared between both foxes. Nucleotide similarity ranged from 93.1 (with genotype 2 of S. lupi and S. vulpis) to 95.8% (with genotype 1 of S. lupi), a higher similarity than noted from sequences of S. lupi from an Andean fox from Peru (91.0 to 93.3%). However, the Poisson Tree Processes for species delineation did not support the existence of a new species Spirocerca. Phylogenetic and nucleotide analyses suggest that these specimens belong to a new variant or genotype of S. lupi or to a cryptic species. Whether the presence of the worms in the stomach has to do with genotypic differences in parasites or host or some combination is uncertain. Spirocerca lupi has never been found in Chilean dogs and must be investigated.

Redescription of Thubunaea schukurovi Annaev, 1973 from Ablepharus chernovi, with notes on the members of Thubunaea (Spirurida: Physalopteridae) from the Palaearctic and Indomalayan realms.

The spirurid nematode Thubunaea schukurovi Annaev, 1973 (Physalopteridae) is redescribed by means of light microscopy and scanning electron microscopy from Ablepharus chernovi collected in Çamliyayla, Turkey. This is the first record of the species from this host and the first record of the genus Thubunaea from Turkey. The taxonomic status of the species of the genus Thubunaea from the Palaeacrtic and Indomalayan realms is reviewed based on their original morphological descriptions; as a result, Thubunaea dessetae Barus & Tenora, 1976, a parasite of Saara hardwickii (Gray) (Agamidae) in Afghanistan, is transferred to the genus Pseudabbreviata as Pseudabbreviata dessetae (Barus & Tenora, 1976) n. comb. Several species described in India, that is, Thubunaea singhi Deshmukh, 1969 and Thubunaea brooki Deshmukh, 1969, both from Hemidactylus brookii Gray (Gekkonidae), and Thubunaea aurangabadensis Deshmukh, 1969 and Thubunaea syedi Deshmukh, 1969, both from Hemidactylus giganteus Stoliczka, are considered members of the genus Physalopteroides; therefore, the following new combinations are proposed: Physalopteroides singhi, Physalopteroides brooki, Physalopteroides aurangabadensis and Physalopteroides syedi. Thubunaea hemidactylae Oshmarin & Demshin, 1972 from Hemidactylus frenatus Duméril & Bibron in Vietnam is considered a member of the genus Physalopteroides as P. hemidactylae (Oshmarin & Demshin, 1972) n. comb.

First report of Thelazia callipaeda Railliet & Henry, 1910 in a zoo lion (Panthera leo) (Linnaeus, 1758) from Romania.

Thelazia callipaeda, the zoonotic "oriental eye worm", is an emerging nematode known to infect a wide range of hosts, particularly carnivores (wild and domestic canids and felids, mustelids, and ursids), but also other mammal groups (suids, lagomorphs, monkeys, and humans), across a wide geographical area. New host-parasite associations and human cases have mostly been reported in endemic areas. A less studied group of hosts is represented by zoo animals that may harbor T. callipaeda.A 23-year old male lion (Panthera leo) that was born in a public zoo in Reșița (Caraș-Severin County, Romania) was submitted to the University of Agricultural Sciences and Veterinary Medicine of Cluj-Napoca for investigations after death from unknown reasons in November 2022. Four nematodes were collected from the right eye during the necropsy and submitted to morphological and molecular characterization.Collected nematodes were morphologically identified as three females and one male T. callipaeda. The BLAST analysis revealed 100% nucleotide identity to numerous isolates of T. callipaeda haplotype 1.The present paper presents a new host-parasite association between T. callipaeda and P. leo from a zoo in Romania.

Molecular characterization of oriental eyeworm (Thelazia callipaeda) detected from raccoon (Procyon lotor) and Japanese raccoon dog (Nyctereutes viverrinus) in Kanto region, Japan.

BACKGROUND: The oriental eyeworm Thelazia callipaeda (Spirurida: Thelaziidae) is an emerging parasitic ocular nematode of carnivores and humans. In domestic animals and humans, the infection causes varying degrees of inflammation and lacrimation, and wild carnivores represent an important reservoir. In this study we examined the infection status and molecular characterization of T. callipaeda in two urban carnivores, raccoons Procyon lotor and wild Japanese raccoon dogs Nyctereutes viverrinus, in the Kanto region of Japan. METHODS: From January 2020 to December 2021, 193 carcasses including 178 raccoons and 15 raccoon dogs were examined for the presence of worms in the eye. The worms from infected animals (one worm per host) were morphologically identified as T. callipaeda. Worms (1-5 worms per host) were subjected to genetic analysis using mitochondrial cytochrome c oxidase subunit I gene sequences. RESULTS: The prevalence of T. callipaeda in raccoons and Japanese raccoon dogs was 20.2% (36/178) and 13.3% (2/15), respectively. The cox1 sequences from 56 worms from 38 animals revealed three haplotypes (h9, h10, and h12). Analysis of multiple worms for five raccoons showed co-infection of two different haplotypes (h9 and h10) in a single host. Comparing our data with published sequences, three sequences obtained from raccoons and raccoon dogs shared the same haplotypes as those reported in humans, dogs, and cats in Japan. CONCLUSIONS: Our findings show a high prevalence of T. callipaeda in raccoons, suggesting that this invasive carnivore species serves as an important natural reservoir of T. callipaeda in the Kanto region of Japan, an area with the highest human population of the country.

Prevention of Thelazia callipaeda Reinfection among Humans.

Thelazia callipaeda is a zoonotic vector-borne nematode that infects and causes eye disease among a wide range of domestic and wild mammals, including humans. We describe an unusual case of reinfection by this nematode in Serbia and call for a focus on preventive measures in endemic areas.

Larvae of Spirocerca lupi and another spirurid species in the same dung beetles: notes on species identification.

Various dung beetle species of the family Scarabaeidae have been reported as intermediate hosts of Spirocerca lupi from different geographical locations, but there has been no data from south-east Asian countries. In the present study in Vietnam, by using morphological and molecular approaches, we identified the third-stage larvae of S. lupi for the first time in two dung beetle species, Catharsius molossus and Copris szechouanicus, adding them as new intermediate hosts of this nematode. In addition, both beetle species were infected with larvae of another spirurid nematode of Physocephalus sp. At large magnifications above 200×, these two spirurid larvae can be differentiated from each other by their body size (1880-2662 vs. 1417-1635 µm) and detailed morphological features of the anterior and posterior ends, such as the length of the buccal cavity, the position of the nerve ring and excretory pore, and tail characters. However, it is difficult to distinguish them at a lower magnification due to their minute size. Moreover, morphometric data of S. lupi larvae vary among reports and the body length may overlap with that of Physocephalus sp., thus, misidentification may occur, indicating the necessity of careful examination for accurate identification. The findings of the present study also supposed that larvae of S. lupi and more spirurid nematodes may be concurrently found in various dung beetle species, including C. molossus and C. szechouanicus, in other south-east Asian countries. Thus, more investigations in various countries should be conducted to identify spirurid larvae from dung beetles in order to fully understand their role as intermediate hosts of S. lupi and other spirurid nematodes.

Spirocercosis and Parvovirus in an Imported Dog: Implications for Biosecurity and Foreign Infectious Disease Risk.

Increased international travel and importation of animals brings with it the potential for spread of infectious diseases. This report details a case in which an 8-month-old male mixed breed dog died shortly after arrival to the United States from complications secondary to spirocercosis, despite having been deemed healthy and approved for international travel. Four weeks following arrival, the dog developed mild tachypnea and diarrhea. Physical examination revealed moderate pyrexia with generalized cerebellar ataxia, moderate anemia, a mature neutrophilia, and severe panhypoproteinemia. Packed red blood cells were administered. The dog was diagnosed with pyothorax and decompensated with septic shock shortly thereafter. Necropsy revealed severe transmural esophagitis with intralesional Spirocerca lupi. Evaluation of the brain revealed cerebellar hypoplasia, and polymerase chain reaction of brain tissue was positive for canine parvovirus. Despite receiving a clean bill of health in its country of origin, this patient ultimately had evidence of 2 different infectious processes that pre-dated its arrival into the United States. While neither of these diseases posed a significant public health risk, this case highlights the role veterinarians play before and after international travel as important barriers against the spread of exotic diseases, and emphasizes that maintaining vigilance is paramount to that task.

Thelazia lacrymalis in horses from Romania: epidemiology, morphology and phylogenetic analysis.

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.

Efficacy of a formulation of sarolaner/moxidectin/pyrantel (Simparica Trio®) for the prevention of Thelazia callipaeda canine eyeworm infection.

BACKGROUND: 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 Trio®) administered orally at monthly intervals, for the prevention of establishment of infection with T. callipaeda in naturally infected dogs. In this formulation, moxidectin is expected to have efficacy against eyeworms, whereas sarolaner and pyrantel are not. METHODS: The 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; Simparica®), and a treatment group (G2; n = 63) in which animals were treated monthly with Simparica Trio (sarolaner/moxidectin/pyrantel) from day 0 to day 150. In total, nine animals were withdrawn from the study (two animals became positive at day 30, and seven for reasons unrelated to eyeworm infection), resulting in 116 animals (n = 58 for G1; n = 58 for G2). RESULTS: In 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). CONCLUSIONS: 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.

Phortica oldenbergi (Diptera: Drosophilidae): A new potential vector of the zoonotic Thelazia callipaeda eyeworm.

Thelazia callipaeda is a zoonotic nematode parasitizing the eyes of many hosts species, primarily dogs. To date Phortica variegata and Phortica okadai are the only known vectors of this nematode in Europe and China, respectively. In this study we investigated the role played by a third species, Phortica oldenbergi, as vector of T. callipaeda in Europe. Drosophilid flies of this species were collected in central Italy and maintained in laboratory. One hundred forty P. oldenbergi were experimentally infected with T. callipaeda L1 recovered from field collected gravid females belonging to haplotype 1, which is that detected in several countries in Europe. Seventy-four (i.e., 60 females and 14 males) specimens died at 5 days post infection (d.p.i.) (±1) and scored negative for T. callipaeda larvae at the dissection. Sixty-six (i.e., 46 females and 20 males) P. oldenbergi survived and were dissected at 21 d.p.i. From those, T. callipaeda L3 were detected in the proboscis of two females (3.0%). Overall, at the molecular analysis, 11.4% (n = 16/140; 13 females and 3 males) scored positive for the presence of T. callipaeda DNA. Data herein reported brings further insights on the biology of T. callipaeda by adding P. oldenbergi as a new potential intermediate host under experimental conditions. The role of this drosophilid in the transmission cycle of T. callipaeda needs to be confirmed under natural conditions.

Subcutaneous spirocercosis in a dog from Costa Rica and first molecular confirmation of Spirocerca lupi in Central America.

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 subcutaneous 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.

An annotated checklist of the genus Pterygodermatites Wedl, 1861 (Nematoda: Rictulariidae), with notes on hosts and geographical distribution.

Species of rictulariid nematodes belonging to the genus Pterygodermatites Wedl, 1861 are parasites of mammals with a wide geographical distribution. The taxonomy of this speciose genus is complex and based mainly on morphological traits of the cephalic structures and cuticular processes. Currently, the known species are allocated to five subgenera [P. (Paucipectines), P. (Neopaucipectines), P. (Multipectines), P. (Pterygodermatites) and P. (Mesopectines)], based on the classification proposed in 1969 by Quentin. Studies on life cycles, taxonomy, biogeography and association of these parasites with mammals peaked at the end of the 1960s. Although the momentum was lost, new species and geographic records have been added in the last 50 years. Therefore, an updated checklist of species of Pterygodermatites is presented here, based on a literature survey. Information on classification, geographical distribution and host range obtained for 68 species is presented, including a distribution map. Moreover, the main differential morphological traits related to each species are tabulated. More than 140 species from 10 different mammalian orders (Carnivora, Chiroptera, Didelphimorphia, Erinaceomorpha, Macroscelidae, Microbiotheria, Paucituberculata, Primates, Rodentia and Xenarthra) have been reported as hosts of Pterygodermatites spp. The taxonomic information here compiled can contribute to future studies on the biogeography, in addition to evolutionary and ecological aspects of this diverse but poorly known genus of parasitic nematodes of mammals.

CHARACTERIZATION OF LESIONS INDUCED BY SPIROCERCA VULPIS (SPIRURIDAE: SPIROCERCIDAE) IN RED FOXES (VULPES VULPES).

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.

Morphological and molecular characterization of Paraleptus chiloscyllii Yin & Zhang, 1983 (Nematoda: Physalopteridae) from the brownbanded bambooshark Chiloscyllium punctatum Müller & Henle (Elasmobranchii: Orectolobiformes).

Paraleptus (Spirurida: Physalopteridae) is a small genus of nematodes, parasitic in fishes, most species of which are inadequately described. Genetic data for these congeners have not been reported. The detailed morphology of P. chiloscyllii was studied using light and scanning electron microscopy, based on newly collected specimens from the brownbanded bambooshark C. punctatum Müller & Henle (Elasmobranchii: Orectolobiformes) in the Taiwan Strait. Some previously unreported morphological features of taxonomic significance were observed, i.e., pseudolabium with two sublateral rows of 6-7 small spines, 7-8 small spines on each lower rim between pseudolabia, deirids not distally bifurcated, vulva with remarkable protruding lip, presence of 1 pair medio-ventral precloacal papillae and 1 pair of discoid protrusions of postcloacal lip in male. The specimens described by González-Solís & Ali's (2015) as P. chiloscyllii from the Arabian carpetshark C. arabicum off Iraq are considered a new species, for which the name P. moraveci n. sp. is proposed. The genetic characterization of the partial small (18S) and large (28S) ribosomal DNA, and the partial mitochondrial cytochrome c oxidase subunit 1 (cox1) of P. chiloscyllii are provided for the first time. There was no intraspecific nucleotide divergence detected in the 18S and 28S regions among different individuals of P. chiloscyllii, but a low level of intraspecific genetic variation was found in the cox1 (0.62-0.92%). The present genetic data is very important for molecular identification, and will be valuable for further invertigantions on the population genetics and phylogeny of this group.

Thelazia callipaeda in Slovakia - From sporadic cases to endemic areas.

Spirurid nematode Thelazia callipaeda, transmitted by the fruit fly Phortica variegata, is a causative agent of an ocular parasitic disease called also canine thelaziosis. Dogs, cats, and wild canids are considered the primary definitive hosts for the parasite, but humans may also serve as aberrant definitive hosts. For long decades the geographic range of T. callipaeda was strictly limited to the territory of Asia, but after the year 2000, the parasite began to spread rapidly through Europe. The first autochthonous infections of dogs and foxes in Slovakia were recorded in 2016. In the present study, the results of a whole-area surveillance for canine thelaziosis are reported. Altogether, 142 cases of infection caused by T. callipaeda were diagnosed by veterinarians in dogs between 2016 and the first quarter of 2021, and two cases of feline thelaziosis were recorded. The majority of the dogs showed mild ocular signs manifested by conjunctivitis; 8.5% of them suffered from more serious mucopurulent discharge, and in two dogs corneal ulceration was recorded. The screening revealed increasing trends in the occurrence of canine thelaziosis from both a temporal and spatial point of view and unambiguously confirms the endemic status of T. callipaeda in Slovakia with the prospect of its further expansion.

Identification of the intermediate host of Gongylonema sp., the etiological agent of the necrotic oropharyngeal disease of the Scops owl (Otus scops).

Since 1997, fledgling Scops owls (Chordata: Strigidae) have been brought to the Brinzal Owl Rescue Centre (Madrid, Spain) with severe lesions in their oral cavities. Lesions consist of the presence of proliferative necrotic material in the oral cavity resulting in white plaques, which can lead to death by starvation. This disease has been detected in owls only within the limits of the city of Madrid. The etiologic agent has been identified as Gongylonema sp. (Nematoda: Spirurida), a nematode genus that includes a coprophagous arthropod as intermediate host in its cycle. The aim of this study was to identify the intermediate host of the parasite. Our work was structured in four component phases: i) Diet study of newborn chicks; ii) trapping arthropods that could be intermediate hosts; iii) molecular detection of the parasite in the selected arthropods: and iv) molecular characterization of the detected parasites by amplifying the cox1 gene. Four male owls were radio-tagged in order to locate their nests and a camera trap was placed to identify the prey brought to the owlets. Secondly, the arthropods present in the hunting areas of the owls were sampled, identified and analyzed by real time PCR (rtPCR). Only oriental cockroaches, B. orientalis (Arthropoda: Blattodea), were positive by rtPCR detection of Gongylonema sp. (66.7%). The nematodes obtained from cockroaches had a 99.8% identity of the cox1 gene with the Gongylonema sp. isolated for the first time in a Scops owl. Furthermore, these sequences only showed an <89% identity with all the other Gongylonema sequences available in the GenBank database. We conclude that the oriental cockroach should be considered as an intermediate host of the etiologic agent of NOD.

Zoonotic Thelazia callipaeda eyeworm in brown bears (Ursus arctos): A new host record in Europe.

Thelazia callipaeda (Spirurida, Thelaziidae) eyeworm, a zoonotic parasite transmitted by lachryphagus drosophilids, causes subclinical to clinical ocular disease in a wide range of vertebrates, including humans. In the past 20 years, the infection spread primarily in areas where the vector thrives, and now is endemic in many European countries. Recently, this nematode has been reported also in the United States. Here, we describe the first record of the brown bear (Ursus arctos) as a host of T. callipaeda. Eight (26.7%) out of 30 bears were found to be infected with T. callipaeda adult worms. In addition, the parasite was detected in 13 wolves (Canis lupus) sharing the same environment with the brown bears. At the molecular characterization, all nematodes belonged to the haplotype 1, which is the only one recorded in Europe, as yet. This finding demonstrates that brown bears may act as hosts for this parasite, suggesting its potential role in the maintenance of the cycle not only in the wild but also in captivity. Under the above circumstances, the brown bear may represent a potential zoonotic risk for humans, both in rural and urban areas where other reservoirs may be infected.