[Ocular thelaziosis in a dog in Germany - an autochthonous case?] / Okuläre Thelaziose bei einem Hund in Deutschland ­ ein autochthoner Fall?

Thelazia callipaeda is a vector-borne nematode transmitted by Phortica spp. (fruit flies) and one of the causes of mild to severe conjunctivitis and keratitis in dogs, cats, rabbits and humans. It has been formerly known as the oriental eye worm based on its geographic occurrence. By now, it has been shown to be endemic in several southern and eastern European countries as well as extending its geographic distribution further throughout Europe. In the present case report, T. callipaeda infection was diagnosed in a female dog from Germany. The dog was referred by a local veterinarian due to a treatment-resistant conjunctivitis. A comprehensive ophthalmologic examination revealed 3 adult eye worms in the conjunctival sac as well as on the bulbar side of the nictitating membrane of the left eye. These were identified by morphological features and molecular techniques as T. callipaeda and represented the primary cause of the dog's unilateral blepharospasm, hyperemic conjunctiva and epiphora. Treatment consisted of manual collection of the adult worms and the administration of moxidectin/imidacloprid as spot on (Advocate®, Bayer). All clinical signs resolved within one week after treatment. In Germany, ocular thelaziosis still represents a rare disease. Usually, it is diagnosed in cats and dogs either imported from abroad or accompanying the owners on holiday travels to endemic countries. The dog presented in this case report was born in Germany. Except for a one-week stay in the Netherlands 11 months before the symptoms began, it had never traveled abroad. Considering the prepatency of T. callipaeda, an autochthonous transmission in this case is highly probable.

Suspected autochthonous Thelazia callipaeda infection in a dog in northern Germany.

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.

Haematobium irritans and Haematobium titillans as potential vectors of Parabronema skrjabini in camels (Camelus bactrianus) in Inner Mongolia, China.

Parabronema skrjabini is one of the most harmful nematodes to camels and is responsible for economic losses in animal husbandry industry. There is an urgent need for in-depth studies of potential vectors of the nematode due to its scant regarding information. As previous studies indicated that flies may be the vectors of P. skrjabini, we captured flies in the main camel-producing areas of Inner Mongolia. After autopsy of the specimens of two species of horn flies, we observed the morphology of the suspected nematode larvae found in them. Internal transcribed spacer ribosomal-DNA gene sequences were considered the best candidate to confirm the species of the larvae found. Our results showed that the homology compared with P. skrjabini was 99.5% in GenBank. Subsequently, we preliminarily identified two species of horn flies through morphological observation and then sequenced the mitochondrial-DNA-gene cytochrome oxidase subunit I obtained from two species of horn flies, with 100 and 99.2% similarity to sequences deposited in GenBank, respectively. Thus, we identified Haematobia titillans and Haematobia irritans and provided evidence for their potential role as vectors of parabronemosis. Our study provides reference for future research on the life history of the nematode and the vectors of parabronemosis.

Vertical Transmission of a Nematode from Female Lizards to the Brains of Their Offspring.

Parasites have evolved a diversity of lifestyles that exploit the biology of their hosts. Some nematodes that parasitize mammals pass via the placenta or milk from one host to another. Similar cases of vertical transmission have never been reported in avian and nonavian reptiles, suggesting that egg laying may constrain the means of parasite transmission. However, here we report the first incidence of transovarial transmission of a previously undescribed nematode in an egg-laying amniote, the common wall lizard (Podarcis muralis). Nematodes enter the developing brain from the female ovary early in embryonic development. Infected lizard embryos develop normally and hatch with nematodes residing in their braincase. We present a morphological and molecular phylogenetic characterization of the nematode and suggest that particular features of lizard biology that are absent from birds and turtles facilitated the evolutionary origin of this novel life history.

Molecular Identification of Oxyspirura Petrowi Intermediate Hosts by Nested PCR Using Internal Transcribed Spacer 1 (ITS1).

Recently, the heteroxenous eyeworm, Oxyspirura petrowi, has gained attention due to its prevalence in the declining game bird, Northern bobwhite (Colinus virginianus), but the intermediate hosts of many nematodes remain unknown. However, identifying the intermediate host of O. petrowi with traditional techniques would be difficult and time-consuming, especially considering there are more than 80 potential orthopteran hosts just in Texas. To screen a large number of samples quickly and effectively, primers for nested PCR (nPCR) were developed using the internal transcribed spacer 1 (ITS1) region. Then the nPCR was used to identify which of the 35 species collected from the Order Orthoptera were potential intermediate hosts of O. petrowi. With this technique, 18 potential intermediate hosts were identified. Later, we collected live specimens of species that tested positive to confirm the presence of larvae, but larvae were not found in the live specimens, nor in the extra tissue of the species that had tested positive for O. petrowi DNA. Despite this, this study demonstrated that nPCR is more sensitive than traditional techniques and can be a valuable tool in determining the intermediate hosts of parasites.

The first autochthonous case of feline ocular thelaziosis in Austria.

Over the last 30 years, Thelazia callipaeda (Spirurida: Thelaziidae) has increasingly been reported as an agent of ocular infections in animals and humans throughout Europe. Following the cases of canine ocular thelaziosis recently recorded in Austria for the first time, in the present paper, we describe the first case of T. callipaeda infection in an Austrian cat with no history of traveling abroad. This finding further supports the occurrence of the parasite's autochthonous transmission cycle in the country. The cat showed serous ocular discharge, conjunctival hyperemia, and mild conjunctival edema in the right eye. Mechanical removal of the parasite from the cat's eye, in combination with milbemycin oxime/praziquantel oral treatment and topical use of tobramycin/dexamethasone eye drops led to complete resolution of the clinical signs within 2 weeks. Results presented in the current study are of great importance for the local veterinarians who seemed largely unaware of this zoonotic parasite. Therefore, increased awareness of medical and veterinary communities is imperative for preventing further infections in both animals and humans.

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.

Predicting the distribution of Phortica variegata and potential for Thelazia callipaeda transmission in Europe and the United Kingdom.

BACKGROUND: Male fruitflies Phortica variegata (Drosophilidae, Steganinae) are the intermediate host of the zoonotic nematode Thelazia callipaeda (Spirurida, Thelaziidae). More than 10 years ago, when T. callipaeda was confined to remote regions of southern Italy, ecological niche models were used to predict the potential distribution of P. variegata across Europe and the likely risk of the nematode spreading through infected dogs travelling to/from endemic regions. As predicted, over the last 10 years T. callipaeda has spread rapidly across Europe. Recently, we identified the potential for its introduction to the UK through infected dogs travelling to/from endemic regions of mainland Europe. METHODS: Here updated information is used to re-evaluate the model-predicted European, and specifically, UK distribution to determine the likelihood of T. callipaeda becoming established. Additionally, the UK distribution of P. variegata was further investigated through snapshot fly trapping at model-predicted locations. RESULTS: Ecological niche modelling using Genetic Algorithm for Rule-set Prediction (GARP) analysis suggests a European range similar to that described previously, with some indication of potential spread further eastward. Finer scale UK mapping suggested that P. variegata presence was limited mostly to southern England, but highlighted regions where P. variegata has not been documented previously. The arbitrary fly trapping identified activity of P. variegata at two locations where the species has been found previously late in the season. No specimens were collected at model-predicted locations, although habitat suitable for the species was identified. CONCLUSIONS: GARP-model prediction of P. variegata distribution suggests presence of suitable conditions in previously undocumented regions of the UK and Europe and highlight the possibility for further spread of T. callipaeda across Europe, including the UK. Further work to validate the P. variegata UK model with field data will help improve its accuracy in predicting suitable areas, whilst surveillance of sylvatic definitive host species in such locations is advised to monitor for evidence of autochthonous T. callipaeda transmission.

Detection of Thelazia callipaeda in Phortica variegata and spread of canine thelaziosis to new areas in Spain.

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.

Competence of Phortica variegata from the United States as an Intermediate Host of the Thelazia callipaeda Eyeworm.

Over the past 20 years, Thelazia callipaeda (the oriental eyeworm) has become endemic in Europe, infecting domestic and wild carnivores and humans. The vector of this nematode, the fruit fly Phortica variegata, has recently been discovered in the United States, and its vector competence is demonstrated for T. callipaeda in this article, therefore representing a potential new threat for infection of carnivores and humans in the United States.

Spreading of Thelazia callipaeda in Greece.

Thelazia callipaeda (Spirurida, Thelaziidae), the so-called oriental eyeworm, has been increasingly reported as an agent of infection in animals and humans from many European countries. Clinical signs range from subclinical to moderate or severe ocular disorders (e.g., epiphora, photophobia, conjunctivitis, keratitis, ulcers). The disease has been also diagnosed in animals from countries of the Balkan area (e.g., Croatia, Bosnia and Herzegovina and Serbia), but only a single case of canine thelaziosis, presumably autochthonous, was reported in Northern Greece. In this study, we provide robust information of the occurrence of thelaziosis in Greece by reporting autochthonous cases of thelaziosis in dogs (n = 46), cats (n = 3) and in one rabbit (Oryctolagus cuniculus) living in Northern and Central regions of Greece. The occurrence of a single haplotype of the cytochrome oxidase subunit 1 gene confirms that the same zoonotic haplotype of the parasite circulating in Europe is also spreading in Greece. The increased awareness of this parasitosis is crucial to limit the risk of further infections in both humans and animals in European countries.

The larval development of Habronema muscae (Nematoda: Habronematidae) affects its intermediate host, Musca domestica (Diptera: Muscidae).

Although the life cycle of the equid stomach parasite Habronema muscae was disclosed more than 100 years ago, little is known about the effect of the developing nematode larvae in its intermediate host, Musca domestica. In a series of experiments, freshly hatched M. domestica larvae were exposed to H. muscae eggs contained in a faecal sample of a naturally infected horse. In daily intervals, 50 fly larvae were removed and transferred on a parasite-free larval rearing medium where they completed their development. Hatched flies were examined for the presence of Habronema third-stage larvae. In two subsequent control groups, flies spend their entire larval life in contaminated horse faeces and in a parasite-free larval rearing medium, respectively. Out of the 700 fly larvae used in the infection experiments, 304 developed into adult flies of which 281 were infected. The average nematode larval burden rose from 3.6 in the group with the shortest exposure to more than 25 in the groups with the longest exposure. The proportion of larvae that developed into the adult insect fell from 82 % in the uninfected control group to 27 % in the positive control group. The pupae of the positive control group were smaller and lighter than those of the uninfected control group. Lower pupal size and weight in the positive control group as well as a lower insect developing rate might be attributed to the destruction of adipose cells in the maggots by Habronema larvae.

First Record of Paratenic Hosts of the Swimbladder Nematode Anguillicola crassus in North America.

Anguillicola crassus is a non-native parasite of the American eel, Anguilla rostrata. Since being introduced into North America, the nematode has spread rapidly across the range of A. rostrata, but paratenic hosts, which may facilitate parasite dispersion, have yet to be identified in the region. We investigated infection of larval A. crassus in 261 fish specimens belonging to 23 species and 12 orders collected from estuarine habitats in South Carolina (salinities 0-9 ppt) and Nova Scotia (10-18 ppt). A total of 35 fish belonging to 5 species and 3 orders were infected with the third-stage larvae (L3) of A. crassus, providing the first record of paratenic hosts for the parasite in North America. In South Carolina, high prevalence and abundance of the worm were found in spot (Leiostomus xanthurus), silver perch (Bairdiella chrysoura), and highfin goby (Gobionellus oceanicus), and a high prevalence but lower abundance was found in mummichog (Fundulus heteroclitus). In Nova Scotia, 2 nematodes were found in a single specimen of tomcod (Microgadus tomcod). All of the infected species are associated with a benthic lifestyle, and some of them are known to move between estuaries along the coastline. Lower infection rates in Nova Scotia may be associated with lower water temperatures and/or higher salinity of the sampling site. Most of the L3 were found encapsulated in mesenteric tissue around the intestine and stomach. No L4 or pre-adult worms were found. Mean body length of the L3 was smaller than L3 stages found in American eels from Cape Breton. This suggests that development of A. crassus is arrested at the L3 in the 5 fish species reported here, supporting their status as paratenic hosts.

Transmission of the eyeworm Thelazia callipaeda: between fantasy and reality.

BACKGROUND: Thelazia callipaeda is transmitted by Phortica variegata, a drosophilid that feeds on lachrymal secretions of mammals. Scientific information on human thelaziosis is still relatively limited, mainly for physicians and ophthalmologists. Indeed, the literature is full of misleading information on the transmission of T. callipaeda to humans. FINDINGS: A recent paper reported a case of human intraocular infestation in a patient from Karnataka. The information presented in that article as well as in other articles in the international literature is outdated and incorrect in several instances, mostly regarding to the localization of T. callipaeda in the host, its biology and routes of transmission. CONCLUSIONS: Physicians and ophthalmologists should be aware that T. callipaeda is larviparous and transmitted exclusively by secretophagous flies. These flies buzz around the eyes of animals and humans at the daytime, landing on the eyes and releasing the infective larvae on the host conjunctiva. That is the only possible way of transmission of T. callipaeda.

Drosophilidae feeding on animals and the inherent mystery of their parasitism.

Insect evolution, from a free to a parasitic lifestyle, took eons under the pressure of a plethora of ecological and environmental drivers in different habitats, resulting in varying degrees of interactions with their hosts. Most Drosophilidae are known to be adapted to feeding on substrates rich in bacteria, yeasts and other microfungi. Some of them, mainly those in the Steganinae subfamily, display a singular behaviour, feeding on animal tissues or secretions. This behaviour may represent an evolving tendency towards parasitism. Indeed, while the predatory attitude is typical for the larval stages of a great proportion of flies within this subfamily, adult males of the genera Amiota, Apsiphortica and Phortica display a clearly zoophilic attitude, feeding on the lachrymal secretions of living mammals (also referred as to lachryphagy). Ultimately, some of these lachryphagous species act as vectors and intermediate hosts for the spirurid nematode Thelazia callipaeda, which parasitizes the eyes of domestic and wild carnivores and also humans. Here we review the scientific information available and provide an opinion on the roots of their evolution towards the parasitic behaviour. The distribution of T. callipaeda and its host affiliation is also discussed and future trends in the study of the ecology of Steganinae are outlined.

Incidence of Cercopithifilaria bainae in dogs and probability of co-infection with other tick-borne pathogens.

BACKGROUND: Cercopithifilaria bainae is a filarioid parasite that infects dogs, being transmitted by Rhipicephalus sanguineus group ticks in many countries of the Mediterranean basin. This study assessed the incidence density rate (IDR) of infection by C. bainae in dogs and the probability of co-infection with other tick-borne pathogens (i.e., Anaplasma platys, Babesia vogeli and Hepatozoon canis), in an area of high endemicity in southern Italy. METHODOLOGY/PRINCIPAL FINDINGS: From March 2011 to October 2012, a field study involving 58 young dogs naturally exposed to tick infestation was conducted. Skin and blood samples obtained from each dog six times during an 18-month period were tested for C. bainae by parasite detection within skin snip sediments, with subsequent confirmation through PCR and DNA sequencing. Dogs examined monthly for ticks and A. platys, B. vogeli and H. canis were microscopically and/or molecularly diagnosed and after the first and the second summer seasons, the IDR for positive animal-month at risk was 3.8% and 1.7% in November 2011 and October 2012, respectively. All 58 C. bainae-infected dogs were simultaneously infected with at least one other tick-borne pathogen. After the first summer season (assessment in November 2011), a C. bainae-infected dog had a 33% probability of being infected with H. canis or A. platys, whereas after the second tick season (assessment in October 2012) the probability of co-infection was 78%, 22% and 11% for H. canis, A. platys and B. vogeli, respectively. CONCLUSIONS: Our data indicate that tick-infested dogs are at risk of acquiring infection by C. bainae. In addition, the detection of C. bainae microfilariae indicates a prior tick exposure and, should stimulate testing for other tick-borne disease causing pathogens.

Tick vectors of Cercopithifilaria bainae in dogs: Rhipicephalus sanguineus sensu lato versus Ixodes ricinus.

Recently, dermal microfilariae of a Cercopithifilaria species (Spirurida, Onchocercidae), namely Cercopithifilaria bainae , were detected in dogs from several geographical areas of the Mediterranean basin. Evidence from both laboratory and field studies support the role of the brown dog tick, Rhipicephalus sanguineus sensu lato, as an intermediate host of this nematode. In the present study, we investigated the competence of Ixodes ricinus nymphs as vectors of C. bainae. On November 2012, fully engorged nymphs of I. ricinus (n = 174) and R. sanguineus s.l. (n = 10) were collected from a dog infected by C. bainae. The presence of C. bainae in I. ricinus was assessed by both microscopic dissection of specimens and detection of nematode DNA (PCR), at days 3, 10, 20 and 30 (T1-T4) post-collection; due to the small number of specimens available, R. sanguineus s.l. were examined using the same methods at T4 only. No developing larva of C. bainae was detected in I. ricinus specimens at different time points (T1-T4), even if four of these specimens were PCR-positive at T1. Seven out of ten R. sanguineus s.l. were positive for C. bainae third-stage larvae (L3) at both microscopical and molecular analysis at T4. This study indicates that C. bainae does not develop in I. ricinus nymphs, which preclude the role of this tick as an intermediate host of this parasite. Data presented herein provide new insights into the biology of this filarioid species and will lead to a better understanding of the role of different tick species as vectors of nematodes.

Arthropod intermediate hosts of Abbreviata antarctica (Nematoda: Physalopteridae) in Australia.

This study examines potential arthropod intermediate hosts for the nematode Abbreviata antarctica. Five species of arthropod (tropical native cricket Teleogryllus oceanicus; native roach Drymoplaneta communis; native termite Nasutitermes exitiosus; and 2 introduced species, the East African roach Naupheta cinerea and the Asian cricket Acheta domesticus) were fed feces from a skink, Tiliqua rugosa, containing embryonated eggs of A. antarctica . The insects were dissected at 4 intervals (1 day, 4 days, 8-16 days, and 25-26 days). Viable third-stage larvae were recovered from cysts on the external wall of the midgut and hindgut of 62.5% nymphal and adult T. oceanicus crickets at 25-26 days post-infection and from a single nymphal A. domesticus cricket. No roaches contained eggs or first-stage larvae after 48 hr, and neither eggs nor larvae were found in termites.