Altered neuronal activity in the visual processing region of eye-fluke-infected fish.

Fish, like most vertebrates, are dependent on vision to varying degrees for a variety of behaviours such as predator avoidance and foraging. Disruption of this key sensory system therefore should have some impact on the ability of fish to execute these tasks. Eye-flukes, such as Tylodelphys darbyi, often infect fish where they are known to inflict varying degrees of visual impairment. In New Zealand, T. darbyi infects the eyes of Gobiomorphus cotidianus, a freshwater fish, where it resides in the vitreous chamber between the lens and retina. Here, we investigate whether the presence of the parasite in the eye has an impact on neuronal information transfer using the c-Fos gene as a proxy for neuron activation. We hypothesized that the parasite would reduce visual information entering the eye and therefore result in lower c-Fos expression. Interestingly, however, c-Fos expression increased with T. darbyi intensity when fish were exposed to flashes of light. Our results suggest a mechanism for parasite-induced visual disruption when no obvious pathology is caused by infection. The more T. darbyi present the more visual stimuli the fish is presented with, and as such may experience difficulties in distinguishing various features of its external environment.

Confirmed cases of human Onchocerca lupi infection: a systematic review of an emerging threat.

Diverse Onchocerca species are present mostly parasitizing ungulates, with the exception of Onchocerca volvulus (O. volvulus) in humans and O. lupi in canids and cats. The human cases due to the O. lupi have been more highlighted during last years. So, the present review was performed to determine the detailed characteristics of confirmed human O. lupi case reports documented worldwide. Hence, a systematic search was done using English international databases (Scopus, PubMed, Web of Science, Embase, ProQuest, and Google Scholar). Totally, 14 confirmed human cases were documented during the last decade, mostly from the USA and Turkey with 7 and 3 cases, respectively. Most cases (7 individuals) were male with the age range of 22-month-old to 54-year-old. The parasite was frequently isolated from the right eye (5 cases), followed by the left eye (4 cases), cervical spinal canal (3 cases), scalp, and right forearm (one case each). Molecular identification of the isolated agent was the preferred way of diagnosis in most cases (9 records). In conclusion, human O. lupi cases have been more highlighted in recent years, whether due to the improved diagnostics and/or host-switching phenomenon, and both veterinarians and healthcare authorities should be alerted.

First report of Thelazia callipaeda in a free-ranging Iberian wolf (Canis lupus signatus) from Spain.

Thelazia callipaeda (Spirurida, Thelaziidae) is a vector-borne zoonotic eyeworm able to infect a broad spectrum of carnivores. Here, we describe the first case of bilateral infection by T. callipaeda in the eyes of an adult female Iberian wolf (Canis lupus signatus) in central Spain. Nematodes collected were morphologically identified (n = 42), and two specimens were molecularly characterized. At the sequence analysis of the partial mitochondrial cytochrome c oxidase subunit 1 gene, T. callipaeda haplotype 1 (the only haplotype circulating in Europe) was detected. The role of the Iberian wolf as a natural reservoir for T. callipaeda in the life cycle of this emerging zoonosis and the implications in conservation are discussed.

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.

Risky business: influence of eye flukes on use of risky microhabitats and conspicuousness of a fish host.

A prerequisite for a parasitic manipulation to be considered adaptive is that it confers a fitness benefit to the parasite, such as increased transmission to another host. These manipulations can involve alterations to a wide range of host phenotypic traits, including microhabitat choice. Eye flukes of the trematode family Diplostomidae use fish as intermediate hosts and must be transmitted by predation to a piscivorous bird. In New Zealand, the diplostomid Tylodelphys darbyi infects the eyes of a widespread endemic freshwater fish, the common bully Gobiomorphus cotidianus. Within the eye, T. darbyi metacercariae achieve large sizes and move freely about the aqueous and vitreous humors of the eye. We hypothesized that higher intensities of T. darbyi would (i) cause bullies to show increased activity and spend more time moving about in open space (i.e., more conspicuous, risky microhabitat) and (ii) reduce their ability to compete for shelter with fish harboring lower infection levels. Our experiments showed that heavily infected fish were more active and spent more time in the open, although the effect was age-dependent, with immature fish displaying decreases in activity and time spent in the open with increasing intensities of infection. We also demonstrated that heavily infected female bullies have a lower probability of using shelter, but males show the opposite pattern. It is possible that using more risky microhabitats increases the likelihood of the fish being eaten by the parasite's predatory avian definitive hosts. However, our findings indicate that age- and sex-dependent effects call for a more nuanced interpretation.

Ocular Sparganosis: The First Report of Spirometra ranarum in Thailand.

A 22-year-old Thai man from the Northeast region presented with acute eye swelling, itching, and discharge on his left eye. He was suspected of having gnathostomiasis and treated with albendazole and prednisolone for 3 weeks. Nine months later, he was treated with high-dose oral prednisolone for the preliminary and differential diagnoses with thyroid-associated orbitopathy and lymphoma. He had been administered prednisolone intermittently over a few years. Then he developed a painless movable mass at the left upper eyelid and recurrent pseudotumor oculi was suspected. The surgical removal of the mass was performed. A white pseudosegmented worm revealed a definite diagnosis of ocular sparganosis by a plerocercoid larva. Molecular diagnosis of the causative species was made based on the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. Proper technique of extraction and amplification of short fragments DNA from formalin-fixed paraffin-embedded tissue successfully identified parasite species. The result from the sequencing of the PCR-amplified cox1 fragments in this study showed 99.0% sequence homology to Spirometra ranarum. This is the first report of S. ranarum in Thailand.

Morphology and molecules resolve the identity and life cycle of an eye trematode, Philophthalmus attenuatus n. sp. (Trematoda: Philophthalmidae) infecting gulls in New Zealand.

Trematodes of the genus Philophthalmus are cosmopolitan parasites that infect the eyes of birds and mammals. They have the potential to affect the survival of their hosts and a few cases of human philophthalmiasis have occurred worldwide. Adults of known Philophthalmus species have never been recorded from bird hosts in New Zealand, despite their cercarial stage being a focus of various studies. Here, we describe a new species of Philophthalmus infecting New Zealand red-billed and black-backed gulls, Philophthalmus attenuatus n. sp. It is distinguished from other marine species of Philophthalmus by its long, thin body shape, consistently longer vitelline field on the left, and its body reflexed at the ventral sucker. We use molecular methods to complete the life cycle of this species, matching it with the larval stage infecting the mud whelk, Zeacumantus subcarinatus, and present a preliminary cox1 phylogeny. In addition, we comment on the validity of some taxonomic characters used to differentiate species of this genus, discuss potential colonisation routes to New Zealand and comment on the potential for zoonotic infection.

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.

First report of eyeworm infection by Thelazia callipaeda in gray wolf (Canis lupus) from Serbia.

Thelazia callipaeda, originally known as an "Oriental eyeworm," is a small nematode parasitizing the conjunctival sacs of domestic and wild animals and humans. Previous studies conducted in Serbia have reported the eyeworm infections in dogs, cats, and foxes, as well as in a human patient. As the data regarding thelaziosis from wildlife is still scarce, the aim of this study was to investigate the presence of T. callipaeda in gray wolf (Canis lupus) from Serbia. All collected nematodes were morphologically identified as T. callipaeda males (n = 64) or females (n = 225). Molecular characterization, conducted by PCR amplification followed by sequence analysis of partial cytochrome c oxidase subunit 1 gene (cox1), revealed only haplotype 1 of T. callipaeda. The overall prevalence of thelaziosis was 38.1% (8/21). In all positive animals, both eyes were affected, with a total parasitic load ranging from four to 132 worms per animal. Our results indicate the important epidemiological role of wolves as wildlife reservoirs of T. callipaeda, expanding geographic range of infection, as well as intra- and interspecies contact rates, although the role of other wild carnivore species (i.e., foxes and jackals) should be investigated in future studies.

Oleic acid-conjugated silver nanoparticles as efficient antiamoebic agent against Acanthamoeba castellanii.

Acanthamoeba castellanii belonging to the T4 genotype is an opportunistic pathogen which is associated with blinding eye keratitis and rare but fatal central nervous system infection. A. castellanii pose serious challenges in antimicrobial chemotherapy due to its ability to convert into resistant, hardy shell-protected cyst form that leads to infection recurrence. The fatty acid composition of A. castellanii trophozoites is known to be most abundant in oleic acid which chemically is an unsaturated cis-9-Octadecanoic acid and naturally found in animal and vegetable fats and oils. This study was designed to evaluate antiacanthamoebic effects of oleic acid against trophozoites, cysts as well as parasite-mediated host cell cytotoxicity. Moreover, oleic acid-conjugated silver nanoparticles (AgNPs) were also synthesized and tested against A. castellanii. Oleic acid-AgNPs were synthesized by chemical reduction method and characterized by ultraviolet-visible spectrophotometry, atomic force microscopy, dynamic light scattering analysis, and Fourier transform infrared spectroscopy. Viability, growth inhibition, encystation, and excystation assays were performed with 10 and 5 µM concentration of oleic acid alone and oleic acid-conjugated AgNPs. Bioassays revealed that oleic acid alone and oleic acid-conjugated AgNPs exhibited significant antiamoebic properties, whereas nanoparticle conjugation further enhanced the efficacy of oleic acid. Phenotype differentiation assays also showed significant inhibition of encystation and excystation at 5 µM. Furthermore, oleic acid and oleic acid-conjugated AgNPs also inhibited amoebae-mediated host cell cytotoxicity as determined by lactate dehydrogenase release. These findings for the first time suggest that oleic acid-conjugated AgNPs exhibit antiacanthamoebic activity that hold potential for therapeutic applications against A. castellanii.

The First Acanthamoeba keratitis Case of Non-Contact Lens Wearer with HIV Infection in Thailand.

Acanthamoeba keratitis (AK) is a rare sight-threatening corneal infection, often reporting from contact lens wearers. An asymptomatic human immunodeficiency virus (HIV)-infected Thai male without history of contact lens use complained foreign body sensation at his left eye during motorbike riding. He had neither specific keratitis symptoms nor common drugs responding, which contributed to delayed diagnosis. By corneal re-scraping, Acanthamoeba-like cysts were detected by calcofluor white staining and agar culture. The etiological agent obtained from the culture was molecularly confirmed by Acanthamoeba spp.-specific PCR, followed by DNA sequencing. The results from BLAST and phylogenetic analysis based on the DNA sequences, revealed that the pathogen was Acanthamoeba T4, the major genotype most frequently reported from clinical isolates. The infection was successfully treated with polyhexamethylene biguanide resulting in corneal scar. This appears the first reported AK case from a non-contact lens wearer with HIV infection in Thailand. Although AK is sporadic in developing countries, a role of free-living Acanthamoeba as an opportunistic pathogen should not be neglected. The report would increase awareness of AK, especially in the case presenting unspecific keratitis symptoms without clinical response to empirical antimicrobial therapy.

A possible relationship between Thromboxane B2 and Leukotriene B4 and the encapsulation of Dirofilaria repens worms in human subcutaneous dirofilariasis.

Human subcutaneous dirofilariosis has several clinical presentations. Many cases present as subcutaneous nodules, as a consequence of a local inflammatory reaction that encapsulates and destroys the worms. In addition, there are cases in which migrating worms located in the ocular area remain unencapsulated. In the present work, the levels of two pro-inflammatory eicosanoids, thromboxane B2 (TxB2) and leukotriene B4 (LTB4) are analysed by commercial Enzime-Linked immunosorbent assay (ELISA) in serum samples from 43 individuals, 28 diagnosed as having subcutaneous dirofilariasis presenting a subcutaneous nodule, five diagnosed as having dirofilariasis, in which the worms remained unencapsulated in the periphery of the eye, and ten healthy individuals living in a non-endemic area, used as controls. The worms were surgically removed, identifying Dirofilaria repens as the causative agent in all cases, by Polymerase Chain Reaction (PCR). Individuals with nodules showed significantly higher levels of TxB2 and LTB4 than healthy controls, whereas significant differences in LTB4 levels were observed between individuals with unencapsulated worms and healthy controls. It is speculated that the absence of LTB4 may contribute to the fact that worms remain unencapsulated as a part of immune evasion mechanisms.

Influence of an extreme climatic event on bilateral asymmetry and occurrence of eye flukes in perch (Perca fluviatilis) and roach (Rutilus rutilus) from a lake in southeast England.

Eye flukes are common infections of freshwater fish and their occurrence as metacercarial stages may occur non-randomly resulting in an asymmetrical distribution within the host eyes. However, from previous studies the presentation of bias by these trematodes lacks consistency suggesting that congenital asymmetrical effects are unlikely to be the cause and exogenous factors, such as environmental stress, may be more influential. The present study, undertaken over a 4-year period, investigates the impact of an extreme heatwave and drought on the annual bilateral asymmetry and occurrence of two eye fluke species (Diplostomum sp. in the lens and Tylodelphys sp. in the vitreous humour) from perch (Perca fluviatilis) and roach (Rutilus rutilus). The onset of the extreme climatic event resulted in a significant negative effect on the occurrence of the eye flukes. Bilateral asymmetry, which was present within both trematode species and hosts, appeared to be highly variable between eye fluke and fish species and also year of study. However, during the pre-drought period both host species demonstrated significant asymmetry for Tylodelphys sp. but not for Diplostomum sp. while during the drought this bias was reversed. The potential role of fluctuating asymmetry of fish hosts in structuring the bilateral asymmetry of eye flukes is discussed.

Ocular spirorchiidiosis in sea turtles from Brazil.

The causes of the beaching and death of sea turtles have not been fully clarified and continue to be studied. Mild, moderate and severe lesions caused by spirorchiidiosis have been seen for decades in different organs and were recently defined as the cause of death of a loggerhead turtle. In the present study, eyes and optic nerves were analysed in green sea turtles with spirorchiidiosis and no other debilitating factors. Injuries to the optic nerve and choroid layer were described in 235 animals (90%) infected with spirorchiids. Turtles with ocular spirorchiidiosis are approximately three times more likely to be cachectic than turtles with spirorchiidiosis without ocular involvement.

Oestrus ovis external ophtalmomyiasis: a case report in Burgundy France.

BACKGROUND: External ophtalmomyiasis (EOM) is a zoonosis related to the presence of Oestrus ovis larvae at the ocular level in small ruminants (i.e. ovine, caprine). In humans, EOM is a rare cosmopolitan disorder, mostly described in warm and dry rural areas in patients living close to livestock areas. In metropolitan France (excluding Corsica), EOM is an exceptional disease with less than 25 cases recorded since 1917. CASE PRESENTATION: We report a case of EOM in a 19-years old man in the last week of September 2016 in Burgundy. CONCLUSION: The diagnosis of an EOM in Burgundy, a French region described as cold and humid, is surprising and could be due to a more marked climatic warming during the vegetative season in Burgundy resulting in the implantation of Diptera of the genus Oestrus sp. in this region.

Ocular thelaziosis due Thelazia callipaeda (Spirurida: Thelaziidae) in Romania: first report in domestic cat and new geographical records of canine cases.

Thelazia callipaeda has been regarded in Europe in the last few decades as a "new" agent of an ocular disease in dogs, cats, several wild species, and not least in humans. After the first European description in Italy, in 1989, the parasite has been reported in many other countries, showing a widespread distribution of this parasite. The first case of canine thelaziosis in Romania has been reported in 2015, and it has been followed by other studies confirming the presence of the nematode in dogs or broadening the host spectrum with some wild carnivore species that have been found positive. Despite being considered suitable primary hosts for T. callipaeda, infection with T. callipaeda in domestic cat is rarely reported in Europe, probably due to some behavioral and physical particularities of this host. This study describes the first autochthonous report of T. callipaeda infection in a domestic cat from Romania. Additionally, to these findings, new geographical records of canine thelaziosis are being revealed, confirming the increased risk of this emerging vector-borne zoonosis.

Morphological Studies of Developmental Stages of Oculotrema hippopotami (Monogenea: Polystomatidae) Infecting the Eye of Hippopotamus amphibius (Mammalia: Hippopotamidae) Using SEM and EDXA with Notes on Histopathology.

The present study was performed to observe histopathological effects of Oculotrema hippopotami Stunkard, 1924 infection in the eye of Hippopotamus amphibius, as well as to reveal new details of morphology and structural features of this monogenean and its comparison between 2 age stages of the parasite. This was done using both light and scanning electron microscopy, energy dispersive X-ray analysis (EDXA) and histopathology. The presence of a mixture of different generations (adult and sub-adult) in one host individual is common for Oculotrema Stunkard, 1924 in contrast to Polystoma Zeder, 1800. New metrical and graphical information obtained for adults and sub-adults compared with the previous studies. Here we show the presence of genital papillae in adults, metrical data on the distal part of the vas deferens. SEM micrographs of sperm ejaculatory structures and information about the flattened dorsal side of the body provided for the first time. Histopathological changes, such as necrosis and hemorrhage in host tissues as a result of O. hippopotami attachment structures are described. Structural analysis of different body parts of O. hippopotami of both age groups are also included. We show qualitative differences in the presence of hardening ions (S, P, Ca) in attachment structures (oral and haptor suckers) that increase with the age of the worm. The presence of sub-adults and adults on the same host, together with high levels of infection without high pathogenicity may account for Oculotrema being one of the most successful parasites among the Monogenea.

The eye fluke Tylodelphys clavata affects prey detection and intraspecific competition of European perch (Perca fluviatilis).

Parasites that occupy the eyes of fish have the potential to affect visual perception and consequently alter the host's behaviour, as these organs provide information about their surroundings. In an experimental study, the feeding behaviour of European perch (Perca fluviatilis) infested with the eye fluke Tylodelphys clavata was examined. The results showed that an individual's ability to identify and approach food items was negatively affected by the infestation intensity of T. clavata. Additionally, the foraging success of an individual was reduced in competition with another, less heavily infested conspecific, when the same food resource was exploited. These alterations in the ability to locate food may have important consequences on the feeding strategy of the fish. Furthermore, the impaired feeding capability caused by T. clavata may also increase the predation risk as heavily infested fish need to spend more time foraging to attain a rate of food intake equivalent to less infested conspecifics.

An adult cystidicolid nematode (Nematoda: Cystidicolidae) from the subcutaneous tissue around the eye of percupinefish, Diodon nichthemerus Cuvier.

Three adult nematode specimens, all ovigerous females belonging to the family Cystidicolidae Skryabin, 1946, were found for the first time in the subcutaneous tissue around the eye of the captive porcupinefish Diodon nichthemerus Cuvier at a public aquarium in Osaka, Japan. Because no male was available, these could not be identified to the genus and species. This case highlights the risk of parasitism in aquaculture puffer fish, as these may ingest small shrimp, which probably act as intermediate hosts for the nematode.