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.

Germination capacity of the Pochonia chlamydosporia fungus after its passage through the gastrointestinal tract of domestic chickens (Gallus gallus domesticus).

This study evaluated the germination capacity of Pochonia chlamydosporia (VC4) fungus after its passage through the gastrointestinal tract of domestic chickens and its interaction with Ascaridia galli and Heterakis gallinarum eggs. Twenty-two domestic chickens were divided in two groups: control group (G1) received shredded corn substrate without VC4; and treatment group (G2) received a single dose of 29 g corn substrate containing 3.3 × 106 conidia/chlamydospores (VC4). Subsequently, chicken fecal samples were collected at intervals of 0, 6, 8, 10, 12, 18 and 24 h. Petri dishes from fecal samples of the treated group (G2) were subdivided (G2a and G2b), and then replicated in 2% agar-water medium for the microbiological test. After VC4 growth, approximately 200 eggs of A. galli (G2a) and H. gallinarum (G2b) were added to each subgroup to evaluation of ovicidal activity. There was fungal viability after passage through chicken gastrointestinal tract and egg predation of 59.9% and 43.2% for A. galli and H. gallinarum, respectively. The present work demonstrates the ability of the fungus P. chlamydosporia to survive after passing through the gastrointestinal tract of domestic chickens, an extreme environment (low pH, enzymes, microbiota and mechanical action), and still germinate after being excreted with feces.

TRANSMISSION OF PTERYGODERMATITES NYCTICEBI IN A COLONY OF GOELDI'S MONKEYS ( CALLIMICO GOELDII) AND EVALUATION OF TREATMENT AND CONTROL.

Over a 2-yr period, four Goeldi's monkeys ( Callimico goeldii) died in a private zoo due to infections with the spirurid nematode Pterygodermatites nycticebi. Therapeutic measures with different anthelmintics were not successful. Due to the severe consequences caused by these infections, different actions were initiated, including sanitation measures and controlling of potential intermediate hosts (coprophagous arthropods). To identify possible intermediate hosts, arthropod species detected in the enclosure-parasite-free German cockroaches ( Blattella germanica), European earwigs ( Forficula auricularia), and rough woodlice ( Porcellio scaber)-were experimentally fed with feces of monkeys with patent P. nycticebi infections, resulting in established infections with third-stage larvae (L3) in roaches and earwigs. Furthermore, spiruroid L3 were detectable in 43% of the roaches and 30% of earwigs caught at the zoo. Polymerase chain reaction and sequence analysis of eggs, larval, and adult stages resulted in identical results, confirming the establishment of the parasite's life cycle in the zoo. This is the first documentation of the vector capacity of the European earwigs for P. nycticebi. As a measure of sanitation, a large part of the enclosure was emptied and cleaned. The Goeldi's monkeys were quarantined and treated with levamisole (7.5 mg/kg sc twice in intervals of 2 wk). Repeated coprologic examinations by zinc chloride flotation were undertaken. After the levamisole therapy, eggs were not found in the feces for 3 mo. However, shortly after resettling the monkeys into the sanitized enclosure, reshedding of small amounts of spirurid eggs was observed, whereupon deworming with levamisole was prescribed several times per year. The sanitation measures and the elimination of the intermediate hosts in a natural enclosure are presented as an example of the long-term controlling of the parasites.

Spirocerca lupi draft genome, vaccine and anthelmintic targets.

Spirocerca lupi is a parasitic nematode affecting predominantly domestic dogs. It causes spirocercosis, a disease that is often fatal. The assembled draft genome of S. lupi consists of 13,627 predicted protein-coding genes and is approximately 150 Mb in length. Several known anthelmintic gene targets such as for ß-Tubulin, glutamate, and GABA receptors as well as known vaccine gene targets such as cysteine protease inhibitor and cytokines were identified in S. lupi by comparing orthologs of C. elegans anthelmintic gene targets as well as orthologs to known vaccine candidates. New anthelmintic targets were predicted through an inclusion-exclusion strategy and new vaccine targets were predicted through an immunoinformatics approach. New anthelminthic targets include DNA-directed RNA polymerases, chitin synthase, polymerases, and other enzymes. New vaccine targets include cuticle collagens. These gene targets provide a starting platform for new drug identification and vaccine design.

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.

Update on the treatment and prevention of ocular thelaziosis (Thelazia callipaeda) in naturally infected dogs from Spain.

This study examines the therapeutic and year-round prophylactic efficacy of different formulations used in dogs in three Spanish areas where canine thelaziosis is endemic. The study was conducted as a Good Clinical Practice, multicentre, randomised field study in privately owned outdoor dogs naturally infected with Thelazia callipaeda. The active pharmaceutical ingredients tested were: an oral formulation of milbemycin oxime 12.5 mg combined with praziquantel 125 mg (A), a subcutaneous sustained-release formulation of moxidectin 10 g (B), a moxidectin 2.5% weight/volume (w/v) spot-on formulation combined with imidacloprid 10% w/v (C), and an eye drop formulation (6 µg) of ivermectin 10 mg/ml diluted 10% in propylene glycol (D). Infected dogs were randomly allocated to treatment Groups A, B, C and D. Dogs testing negative for T. callipaeda inspection in two visits (Day 7/Day 14 and D30) were enrolled in the prophylaxis trial and reallocated to the corresponding study group (A, B, C or D). Treatment efficacy ranged from 70.4% recorded in Group A 1 week after treatment, to 100% recorded in Group C on Day 30 and in Group B on Day 60. Treatment was more efficacious in Group D (85.7% 1 week after treatment) than A, but was never 100% efficacious as in Groups B and C. Year-round prophylactic efficacy was 83.3% in Group A, 100% in Group B, 93.5% in Group C and 87.5% in Group D. In conclusion, products containing moxidectin were highly efficacious both in treating and preventing canine thelaziosis. Milbemycin also emerged as a good option. However, the off-label use of topical or subcutaneous ivermectin should be avoided due to possible adverse reactions such as pruritus, irritation or redness. In endemic areas, monthly prophylaxis to limit the spread of T. callipaeda to new areas across Europe and reduce zoonotic risks is essential.

Thelazia rhodesi in a dairy farm in Romania and successful treatment using eprinomectin.

Bovine thelaziosis can be caused by Thelazia rhodesi Desmarest 1828, Thelazia gulosa Railliet & Henry 1910 and Thelazia skrjabini Erschow 1928 which are known to be transmitted by species of Muscidae. Therapeutic measures in cattle include mechanical removal of the adult parasites, followed by irrigation of the conjunctival sac with different solutions such as levamisole (1%) or administration of ivermectin (0.2 mg/kg SC), doramectin (0.2 mg/kg SC or pour-on) both with high efficacy, however, the use of such macrocyclic lactones can be problematic in lactating cattle. Nematodes harboured in the eyes of some cattle from Romania were morphologically identified as Thelazia rhodesi and the PCR analysis confirmed the diagnosis with a 98.0% nucleotide similarity for ITS1 sequence, with other sequences available in GenBank. The present paper reports the presence of T. rhodesi in cattle from a dairy farm in Romania and the recovery after treatment with eprinomectin. To the best of our knowledge this is the first report on the efficacy of eprinomectin against Thelazia spp. and the first molecular confirmation of T. rhodesi in Romania.

Evaluation of Orally Administered Anthelmintic Treatment Options for Dentostomella translucida in Naturally Infected Mongolian Gerbils (Meriones unguiculatus)

Objective: This study aimed to evaluate the efficacy of eprinomectin, moxidectin and fenbendazole for treating Dentostomella translucida infections in naturally infected Mongolian gerbils (Meriones unguiculatus). Methods: A total of 28 gerbils were placed in individually numbered cages to determine the individual animal parasite load. Eggs per gram (EPG) counts were used to estimate the efficacy of the drugs. The day before the anthelmintic administration was denoted as day 0, and the EPG counts were determined by the McMaster technique from the stool removed from the cage bottom on days 7, 14, 21 and 28. The animals were assigned to one of four treatment groups according to their day 0 EPG counts. The orally administered drugs in the treatment groups were eprinomectin (15 mg/kg), moxidectin (0.4 mg/kg) and fenbendazole (12 mg/kg) for groups 1-3, respectively. The fourth group served as the control (without any drug administration). Results: Treatment efficacy was evaluated based on weekly EPG counts. The values decreased to zero in the fenbendazole group at 4 weeks of follow-up after treatment, and no parasite was found in any of the repeated examinations. The eprinomectin and moxidectin groups exhibited a fluctuating EPG state on both individual and group basis. Conclusion: D. translucida, which is known as the specific parasite of gerbils, can easily affect other members of the animal colony; thus, the control of its presence in gerbil breeding units is necessary. Therefore, the reported effective drug treatments are important for the fight against the investigated parasitic infection.

Feline thelaziosis (Thelazia callipaeda) in Spain: state-of-the-art and first prophylactic trial in cats.

OBJECTIVES: Thelaziosis caused by the nematode Thelazia callipaeda (Spirurida, Thelaziidae) affects the eyes of domestic and wild carnivores, lagomorphs and even humans. The role of cats in spreading thelaziosis remains unclear. The present study assesses the current status of feline T callipaeda infection in Spain and presents the results of the first prophylactic trial conducted in this species. METHODS: We 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). RESULTS: In study 1, 69 Thelazia-infected cats were examined. Autochthonous cases were detected in 18/26 municipalities surveyed in Spain, which corresponds to 88.4% (n = 61/69) of the total number of cases. In study 2, 74 cats (20 from La Vera region and 54 from Orense province) were examined from 2011 to 2013. Ten of these cats (13.5%) were infected with T callipaeda. The infection prevalence was 40% (n = 8/20) in La Vera and 3.7% (n = 2/54) in Orense. The therapeutic efficacy of ivermectin was assessed over a summer (2011) in 12 cats, while that of moxidectin was examined over a 17-month period in seven cats. Four of these seven cats were subsequently included in an all-year-round prophylaxis trial. Treatment efficacy ranged from 91.7% (ivermectin) on day 28 after treatment to 100% (moxidectin) on day 14. Moxidectin prevented reinfections in cats. CONCLUSIONS AND RELEVANCE: Our data confirm the endemic status of T callipaeda infection in Spain, revealing its high prevalence in cats living in known endemic areas. Moxidectin was effective in treating and preventing infection. We strongly recommend the inclusion of thelaziosis in the differential diagnosis of pets and humans presenting with ocular manifestations, along with adequate preventive measures.

Efficacy of a combination of imidacloprid 10%/moxidectin 2.5% spot-on (Advocate® for dogs) in the prevention of canine spirocercosis (Spirocerca lupi).

The nematode Spirocerca lupi is a major canine parasite in warm regions of the world, classically causing parasitic nodules in the esophagus, aortic aneurysms, and spondylitis. This study evaluated the preventive efficacy of monthly treatment with imidacloprid 10%/moxidectin 2.5% spot-on (Advocate® for dogs) administered over a period of 9 months in young dogs naturally exposed to S. lupi on Réunion island. One hundred and twelve puppies, aged from 2.0 to 4.0 months and with a negative spirocerca fecal examination at inclusion, completed the study. They were randomly allocated to two groups. Group A puppies (n=58) received nine spot-on treatments with Advocate® at the minimum dose of 2.5 mg moxidectin/kg bw at monthly intervals. Control group B puppies (n=54) received no treatment for S. lupi. During the study, regular clinical and fecal examinations were performed, as was final upper gastrointestinal endoscopy. Endoscopy showed that 19 dogs from group B had spirocerca nodules, corresponding to a prevalence of 35.2% in dogs aged 12 to 14 months. In contrast, only one dog from group A had a nodule, corresponding to a preventive efficacy of 94.7% (p<0.0001). None of the 378 fecal examinations were positive for spirocerca. This study confirms a high prevalence of canine spirocercosis on Réunion and shows that infestation occurs in very young puppies. Furthermore, it demonstrates that monthly spot-on administration of a combination of imidacloprid 10%/moxidectin 2.5% (Advocate® for dogs) in puppies starting at the age of 2 to 4 months achieves effective and safe prevention of canine spirocercosis.

Systematic review on infection and disease caused by Thelazia callipaeda in Europe: 2001-2020.

Cases of thelaziosis by Thelazia callipaeda have been increasing considerably in Europe throughout the 21st century, with recent emphasis on Eastern Europe. A systematic review was conducted using defined search terms across three major databases and, additionally, with the examination of the references of the 56 articles selected. Available information about epidemiological and clinical features of all cases of thelaziosis by T. callipaeda in companion animals, wildlife and humans was extracted, evaluated and subjected to qualitative and quantitative analysis. In all cross-sectional studies about dogs, cats and red foxes, males were more frequently infected than females (dogs: p = 0.0365; cats: p = 0.0164; red foxes: p = 0.0082). Adult dogs seem to be more prone to infection (p < 0.0001), as well as large-sized dogs (p < 0.0001), and companion animals that live exclusively outdoors (p < 0.0001). Dogs and red foxes involved in these cross-sectional studies harboured significantly more female than male nematodes (p < 0.0001). Thelaziosis by T. callipaeda is far from controlled in Europe. Only through updated epidemiological data, knowledge improvement and awareness can correct diagnosis and appropriate treatment and prevention be ensured to tackle this zoonosis.

TITLE: Revue systématique de l'infection et de la maladie provoquées par Thelazia callipaeda en Europe (2001­2020). ABSTRACT: Les cas de thélaziose par Thelazia callipaeda ont considérablement augmenté en Europe tout au long du 21e siècle, l'accent étant récemment mis sur l'Europe de l'Est. Une revue systématique a été menée en utilisant des termes de recherche définis dans trois grandes bases de données, et aussi en examinant les références des 56 articles sélectionnés. Les informations disponibles sur les caractéristiques épidémiologiques et cliniques de tous les cas de thélaziose par T. callipaeda chez les animaux de compagnie, la faune et les humains ont été extraites, évaluées et soumises à une analyse qualitative et quantitative. Dans toutes les études transversales sur les chiens, les chats et les renards roux, les mâles étaient plus fréquemment infectés que les femelles (chiens : p = 0,0365; chats : p = 0,0164; renards roux : p = 0,0082). Les chiens adultes semblent être plus sujets aux infections (p < 0,0001), ainsi que les chiens de grande taille (p < 0,0001) et les animaux de compagnie vivant exclusivement à l'extérieur (p < 0,0001). Les chiens et les renards roux impliqués dans ces études transversales abritaient significativement plus de nématodes femelles que de mâles (p < 0,0001). La thélaziose par T. callipaeda est loin d'être contrôlée en Europe. Ce n'est que grâce à des données épidémiologiques mises à jour, à l'amélioration des connaissances et à la sensibilisation que l'on pourra corriger le diagnostic et assurer un traitement et une prévention appropriés pour lutter contre cette zoonose.

Thelaziosis due to Thelazia callipaeda in Europe in the 21st century-A review.

Thelazia callipaeda was first described at the beginning of the 20th century in Asia, but this eyeworm is now frequently reported in Europe in the 21st century. To date, thelaziosis has been described in the following European countries (in order of appearance): Italy, France, Germany, Switzerland, Spain, Portugal, Belgium, Bosnia and Herzegovina, Croatia, Serbia, Romania, Greece, Bulgaria, Hungary, Slovakia, the United Kingdom, Turkey and Austria. The infected vertebrate host species include domestic carnivores (dogs and cats), wild carnivores (red foxes, wolves, beech martens, wildcats and golden jackals), lagomorphs (brown hares and wild European rabbits) and humans. In Europe, 11 cases of human thelaziosis have been reported, the majority of which are autochthonous. However, some of them have been imported, a fact which highlights the importance of surveillance policies to restrict cross-border spread of the parasite. The objectives of this article are to review key aspects of the epidemiology of T. callipaeda, summarise animal and human cases in Europe and emphasise the importance of education and awareness among veterinarians, physicians (particularly ophthalmologists) and animal, in order to owners to tackle this zoonosis.

Prevention of canine ocular thelaziosis (Thelazia callipaeda) with a combination of milbemycin oxime and afoxolaner (Nexgard Spectra®) in endemic areas in France and Spain.

In the past decade, canine thelaziosis due to Thelazia callipaeda has been diagnosed in an increasing number of European countries, with endemic areas being identified. A multi-center field trial was conducted in endemic areas in France and Spain to evaluate the efficacy of monthly administrations of the oral milbemycin oxime/afoxolaner combination (NexGard Spectra®) for the prevention of T. callipaeda infection in at-risk dogs. A total of 79 dogs negative for T. callipaeda and with a clinical history of eyeworm infection in the past two years completed the study. Dogs were randomly allocated either to a negative control group (42 dogs) or to the NexGard Spectra® treated group (37 dogs). All dogs were followed up for a 6-month period and assessed monthly for the presence of nematodes on the eyes and for the signs of ocular thelaziosis (e.g., conjunctivitis, keratitis, and ocular discharge). When the presence of nematodes was confirmed, the conjunctival fornix was flushed with a saline solution for parasite recovery and counting, and the dogs were treated appropriately. Recovered parasites were stored in 70% alcohol for subsequent morphological identification. During the course of the study, 57.1% (24/42) of the control dogs were diagnosed positive for Thelazia infection, which illustrates a high incidence rate of parasite infection. Conversely, no eyeworm was recovered from any of the 37 dogs that received NexGard Spectra®. All parasites sampled were confirmed to be T. callipaeda. This clinical field study demonstrated that monthly administrations of NexGard Spectra® provided 100% preventive efficacy against canine thelaziosis.

Therapeutic and prophylactic efficacy of milbemycin oxime (Interceptor) against Thelazia callipaeda in naturally exposed dogs.

Two trials were conducted to evaluate the therapeutic and prophylactic activity of milbemycin oxime (Interceptor, Novartis Animal Health) against the eye-worm Thelazia callipaeda (Spirurida, Thelaziidae) infection. In Trial 1, the therapeutic efficacy of milbemycin oxime was evaluated in 55 naturally infected dogs treated with min. 0.5mg/kg milbemycin oxime. The dogs were clinically examined for the presence of eyeworms before and again 7 days after treatment. Dogs still positive were given a second treatment and re-checked again a week later. Forty-eight of the 55 dogs tested negative 1 week after treatment (87.3% reduction of infection rate). Following the second treatment 6 of these 7 dogs tested negative 1 week later resulting, after two treatments, in a reduction of infection rate of 98.2%. In Trial 2, the prophylactic efficacy of milbemycin oxime was evaluated in 60 uninfected dogs. Thirty dogs were treated with milbemycin oxime monthly from June to November with the recommended dose rate for the prevention of heartworm disease (> or =0.5mg/kg), 30 dogs served as untreated controls. At the end of the trail 1 dog in the treated group and 10 dogs in the control group became infected during the trial. The incidence of infection differed significantly between treated and control dogs (p=0.0056). The efficacy of the prophylactic use of a monthly treatment with milbemycin oxime showed 90% efficacy in reducing T. callipaeda infection rate.

[Parasitic dead-end: update]. / Impasses parasitaires: quoi de neuf?

Parasitic dead-ends occur when a parasite is unable to establish a permanent interaction in an unnatural host. Although the likelihood of successful reproduction by the pathogenic agent is nul, parasitic dead-end heralds capture of new parasites and therefore expansion of the host range. Angiostrongyliasis due to A. cantonensis or A. costaricensis, anisakiasis, Ancylostoma caninum infection, gnathostomiasis and sparganosis are undoubtedly emerging zoonoses of particular medical interest. Prevention of these diseases relies on abstinence from eating raw meat from invertebrates or cold-blooded (poikilotherm) vertebrates (e.g. used in exotic dishes). These guidelines must be included in recommendations to travelers.

Evaluation of the efficacy of monthly oral administration of afoxolaner plus milbemycin oxime (NexGard Spectra(®), Merial) in the prevention of adult Spirocerca lupi establishment in experimentally infected dogs.

The nematode Spirocerca lupi (Rudolphi, 1809) is widely distributed but mostly occurs sporadically with stable populations only in certain geographic areas. This helminth mainly infects dogs and wild canids. Primary pathology relates to migration of third stage larvae (L3) damaging the thoracic aorta and establishment of adults in nodules in the oesophagus. The objective of the present study was to evaluate the efficacy of milbemycin oxime in combination with afoxolaner (NexGard Spectra(®), Merial), administered monthly, in preventing establishment of adult worms after experimental infection. Two groups consisting of eight animals each were experimentally infected with 15 L3 on Days -28, -14 and -2, respectively (45 L3 per animal in total). Group 1 dogs served as untreated (negative) control, whereas animals in group 2 were treated with NexGard Spectra(®) at a minimum dose of 0.5mg/kg milbemycin oxime on Day 0 and from then onwards every 28 days up to Day 140 (six treatment occasions). Endoscopy was performed on Day 112 and for some animals also Day 140. Necropsy for worm recovery and nodule/lesion scoring was performed on Day 168. All eight animals in the control group (group 1) presented with 1-3 nodules and worm counts ranging from 9 to 41. Six animals in the NexGard Spectra(®) group presented with 1-4 nodules and worm counts ranging from 1 to 5. Significantly (p<0.05) fewer worms were collected from treated animals in the treated group (geometric mean 1.7) versus the negative control group (geometric mean 22.0) with 92.3% efficacy calculated. There was no significant (p>0.05) difference between groups with reference to number of nodules in the oesophagus. However, nodules in the control group were significantly (p<0.05) larger than those in the treated group. Number and size of lesions in the dorsal aorta did not differ statistically between groups 1 and 2. Because NexGard Spectra(®) was administered 28 days after onset of inoculation, migrating and developing L3 caused damage to the aorta wall of animals in the treated group. Milbemycin oxime (administered as NexGard Spectra(®)) demonstrated effectiveness in reducing infection with adult Spirocerca lupi worms in the oesophagus.