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Alternatives to conventional chemical treatments for parasitic diseases in fish are needed. Microalgal-sourced fatty acid ethyl esters (FAEEs) have shown an antiparasitic effect against Gyrodactylus turnbulli infection in guppies. Here, we tested a range of commercial FAEEs of various carbon chain lengths and unsaturation levels against two fish parasites. Guppies and barramundi infected with G. turnbulli and Trichodina sp., respectively, were used. The most effective FAEE, after excluding those toxic to fish, was ethyl laurate (12:0). For both parasites, the LD50 was 18.75 µM within 250 min of incubation. Ethyl eicosapentaenoate (20:5n3) was the next most effective FAEE against G. turnbulli, and dihomo-γ-linolenic acid ethyl ester (20:3n6) and ethyl α-linolenate (18:3n3) were the next most effective against Trichodina sp. In addition, FAEEs prepared from the microalga Phaeodactylum tricornutum residue, after fucoxanthin extraction, were examined against Trichodina sp. infection in barramundi for the first time. LD85 and LD100 was achieved at 2.5 and 5 µL mL-1 of the FAEE preparation, respectively. In vivo, immersion of infected barramundi in 1.25 µL mL-1 of this preparation for 24 h reduced infection prevalence from 100% to 53% and was non-toxic to fish.

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BACKGROUND: Parasitic infestations are one of the most significant problems in global aquaculture. In addition to direct economic losses due to significant mortalities, parasites may have a significant negative impact on the fish's behaviour, energetic demands, trophic position, interspecific competition, growth rate, and reproductive function. OBJECTIVES: We conducted the present study to evaluate the infection status of parasitic pathogens in freshwater sutchi catfish (Pangasianodon hypophthalmus) and silver dollar (Metynnis hypsauchen) farmed in Alborz province of Iran. METHODS: Between January and February 2021, 140 ornamental fish, including 70 sutchi catfish (P. hypophthalmus) and 70 silver dollar (M. hypsauchen), were delivered from various ornamental fish farms for parasitological analysis. Thorough macroscopic and microscopic examinations were performed to detect parasitic infections in delivered freshwater ornamental fish. RESULTS: A total of six parasite species, including five protozoan species (Nyctotherus piscicola, Trichodina heterodentata, Ichthyophthirius multifiliis, Protoopalina sp., and Hexamita sp.) and one monogenean species (Ancyrocephalus sp.), were identified in the examined fish. The overall prevalence of the recovered parasites was 46.43% (65/140) of the fish. CONCLUSIONS: In the current study, several parasites, including Hexamita sp., Ancyrocephalus sp., Protoopalina sp., and N. piscicola, were identified as the first records of the sutchi catfish (P. hypophthalmus) and/or silver dollar (M. hypsauchen), and these two fish species are known as new hosts for the isolated parasites in ornamental fish farms in Iran. Assessing the parasitic fauna of ornamental fish is therefore critical for preventing parasite introduction into adjacent provinces as well as neighbouring countries and promoting fish health.

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The thick-shelled river mussel Unio crassus Philipsson, 1788 is a species native to many European habitats, with declining populations. The impact of parasite communities on health status of this species is poorly understood. In this study, parasites of 30 U. crassus specimens from the Our and Sauer Rivers in Luxembourg were identified morphologically and, in some cases, using molecular genetic methods. The findings were correlated to selected parameters (total length, visceral weight, shell lesions, gonadal stage). The 2 populations did not differ in shell length, visceral weight, number of males and females, gonadal scoring, shell lesions, and the occurrence of glochidia. The prevalence and infestation intensities of detected Trichodina sp., Conchophthirus sp., and freshwater mite larvae did not differ between the 2 populations, whereas the prevalence and infestation intensities of mite eggs, nymphs, and adults were significantly higher in the Sauer River. Rhipidocotyle campanula and European bitterling Rhodeus amarus larvae were only present in the Sauer. Histopathology revealed the destruction of the gonads by R. campanula and tissue damage by the mites. The only significant correlation of the selected parameters was a positive correlation between R. amarus occurrence and total length as well as a negative correlation between R. amarus occurrence and gonadal stage. In the Sauer River, 2 mussels were found to be hermaphrodites.

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Mobilids are among the most taxonomically diverse but morphologically uniform groups of epibiotic ciliates. They attach to their hosts by means of an adhesive disc as harmless commensals such as Urceolaria, or as parasites causing significant economic loss such as some Trichodina species. We investigated the diversity, species boundaries, and phylogenetic relationships of mobilids associated with freshwater planarians, using 114 new sequences of two mitochondrial (16S rRNA gene and cytochrome c oxidase gene) and five nuclear (18S rRNA gene, ITS1-5.8S-ITS2 region, D1/D2 domains of 28S rRNA gene) markers. Although the morphological disparity of the isolated trichodinids and urceolariids was low, Bayesian coalescent analyses revealed the existence of five distinct evolutionary lineages/species given the seven molecular markers. The occurrence of mobilids perfectly correlated with their planarian hosts: Trichodina steinii and two Urceolaria mitra-like taxa were associated exclusively with the planarian Dugesia gonocephala, Trichodina polycelis sp. n. with the planarian Polycelis felina, and Trichodina schmidtea sp. n. with the planarian Schmidtea polychroa. Host organisms thus very likely constitute sharply isolated niches that might permit speciation of their epibiotic ciliates, even though no distinct morphological features appear to be recognizable among ciliates originating from different hosts.

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With the aim to determine the prevalence and mean parasite intensity in Oncorhynchus mykiss, 120 specimens were analyzed between April and September 2018. The cavity analysis was done by visual inspection, scraping of mucus, and extraction of the branchial arches. In the evisceration process, the intestine, the liver, and the kidney were separated, while the musculature was analyzed using the "candling table" method. All the collected material was preserved in 10% buffered formaldehyde and sent to the Laboratory of Parasitology and Parasitic Diseases, of the Center for Agroveterinary Sciences (CAV) of the University of the State of Santa Catarina (UDESC) for making the slides and identifying the parasites. Parasites were detected only in the analysis of the scraping of body mucus. Only specimens of the genus Trichodina were identified, in 34 of the 120 fish analyzed, in two collections, resulting in a prevalence of 28.33%. In total, 144 specimens of Trichodina were observed. The overall mean intensity was of 4.24 parasites in each fish analyzed. Characteristic lesions of infection by protozoa were not identified. This is the first report of the occurrence of Trichodina spp. in O. mykiss bred in an intensive system in Brazil, with low rates of parasitic infection in the mountain region of Santa Catarina.(AU)

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Information regarding trichodinid ectoparasites on marine fishes of North America is relatively scarce. In this study, 5 ciliate species from the family Trichodinidae were found associated with 8 fish hosts from the rocky intertidal zone of the western coast of the Baja California Peninsula (BCP), Mexico. All of the host-parasite relationships recorded here are new. Furthermore, 3 of the trichodinid species found are recorded for the first time for Mexico. Trichodinids taxa do not show a noticeable distributional gradient along the BCP, which suggests a wide-continuous distribution of the species throughout the study area.

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Trichodinid ciliates were isolated from calanoid copepods and were characterized by morphological and molecular techniques. The trichodinids found in this study were morphologically similar to Trichodina acuta, but morphometric differences were observed between our specimens and T. acuta. The T. acuta specimens, isolated from fish in several geographical locations around the world, were slightly larger than specimens in our population. In fact, our morphological and morphometric data are more similar to Trichodina diaptomi populations, also found on copepods. Moreover, our phylogenetic analyses revealed that the 18S rRNA gene sequence from our samples emerged in a clade with two T. acuta sequences, infesting fish hosts in China. These three sequences showed 100% identity. Our data, along with a taxonomic review, suggest that the species T. diaptomi, described recently, is actually a synonym of T. acuta and that this species may be related to fish and copepod hosts. In conclusion, we highlight the urgent need to better investigate the polymorphism and host specificity within the Mobilida, as this data may represent important characters to better understand the evolution into this order.

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In the present study, we provide morphological and molecular characterization of two Trichodina species, T. acuta Lom, 1970 and T. funduliWellborn, 1967, isolated from koi (Cyprinus carpio) and loach (Paramisgurnus dabryanus), respectively. Morphological characters of the two Trichodina species were mainly investigated on the basis of dry silver nitrate-impregnated specimens. Both species are medium-sized and possess well-developed denticles comprising strongly sickle-shaped blades, well-developed central parts, and straight rays. Trichodina acuta can be easily distinguished from the other Trichodina species that possess a clear central circle by the well-developed sharp blade apophysis, and the gap between ray tip and central circle. Trichodina funduli is a poorly known species that is easily confused with T. heterodentata Duncan, 1977, however the latter species has thinner denticles. The small subunit ribosomal RNA gene sequences of Trichodina acuta and T. funduli were incorporated into phylogenetic analyses. Our findings suggest that the phylogenetic lineage of trichodinids might not correspond with their living environments, host species or even some morphological characteristics.

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Tilapia is the common name for a wide spectrum of cichlid fishes usually selected for aquaculture activities. However, some aspects of the natural history of these fishes, such as the diversity of certain ectoparasitic protistan groups remain understudied. In order to understand the diversity of ciliates of the family Trichodinidae parasitizing tilapia around the world, a database with all available accounts was assembled. This information, along with records derived from our own recent research, allowed us to generate a checklist containing all the records for tilapia-Trichodinidae associations. The checklist is presented as a host-parasite list containing 44 nominal taxa from 29 countries on four continents, and it also presents the first data from Argentina, where tilapia culture is still an incipient, but growing activity. The observed high species richness of trichodinids is probably derived from a set of specific taxa for tilapia, along with an important component of species acquired from the new environments where tilapia have been introduced. Data presented here could be used along with existing information of metazoan parasites to build a general view about diseases that affect tilapia.

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During a survey of trichodinids in Chongqing, China, two populations of Trichodina reticulata Hirschmann & Partsch, 1955 were isolated from gills of the freshwater fishes, Carassius auratus and Ctenopharyngodon idellus, and 11 molecular samples containing small subunit (SSU) rDNA and internal transcribed spacer (ITS)-5.8S rDNA were newly sequenced. Both populations fell within the range of morphometry and corresponded well with the diagnostic morphological features of Trichodina reticulata Hirschmann & Partsch, 1955. It should be noted that one population possessed obvious central granules in the adhesive disc, while the other one did not. The detailed morphological redescription for these two populations is given in this work. Although some intraspecific differences were found, both populations were confirmed as Trichodina reticulata based on the morphological study and molecular data, including the sequence analysis of the primary and secondary structures of SSU rDNA, genetic distances and phylogenetic tree. Our study supports the assertion that central granules in the adhesive disc might be an auxiliary feature rather than a heritable character for determining taxonomic affiliations among trichodinids.

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The present study performed three experiments to establish a practical prevention strategy for the ectoparasitic flagellate Ichthyobodo salmonis and ciliate Trichodina truttae in hatchery-reared juvenile chum salmon Oncorhynchus keta using dietary supplementation with oregano essential oil. Experiment 1 showed that a diet supplemented for 3 weeks with 0.02% oregano essential oil significantly prevented infection with I. salmonis and T. truttae in juveniles reared in small tanks. Experiment 2, in outdoor hatchery ponds, demonstrated that the oregano treatment completely prevented I. salmonis infection for 52 days and T. truttae infection for 38 days. Oregano-treated juvenile mortality attributable to infection with these protozoans also decreased to 7.6% of control juvenile mortality, confirming the utility of this treatment in cultured O. keta. Physiological analyses of the oregano-treated juveniles elucidated the treatment's safety in relation to their metabolism, osmoregulation, natural immunity and olfactory responses and also detected carvacrol (a major component of oregano essential oil which shows antimicrobial activity) on the skin. In experiment 3, exposure of the two protozoans to oregano essential oil revealed a weak antiparasitic action on the body surface of the juvenile O. keta. The overall results demonstrate that dietary oregano supplementation is a practical prevention strategy for I. salmonis and T. truttae in hatchery-reared juvenile O. keta and suggest the possibility that its anti-parasitic action is attributable to a component of the oil that emerges onto the skin of the body of the fish.

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Trichodinids are peritrichous ciliated protozoa that affect both wild and cultured fishes. Several Trichodina species have low host specificity and are morphologically distinct, facilitating their identification based primarily on the presence of adhesive discs and the number of attached denticles. A trichodinid species named Trichodina compacta was first reported by Van As and Basson (1989) (Protozoa: Ciliophora: Peritrichia). However, in trichodinid infestations, morphological characteristics are insufficient for identifying the infesting species. Therefore, molecular and phylogenetic analyses are considered to be promising and useful tools for identifying the infesting species. This study aimed to achieve the molecular identification of a trichodinid infestation in Nile tilapia and to construct the phylogenetic relationships between the identified species and other peritrichous parasites. Moreover, we also aimed to study the pathological and immunological impacts of trichodinids on fry tissue to improve our understanding of the immune responses of teleost fish to trichodinae parasitic infestations and develop a better control method. Here, we used molecular techniques to identify the isolated trichodina species as T. compacta and demonstrated that Trichodina infestation in Nile tilapia is associated with remarkable immunogenic and inflammatory responses (increased il-1ß expression and decreased il-8 and tgf-ß expression). These findings improve our understanding of the responses of teleost fish to trichodinid parasite infestation and will be helpful for the development of novel control strategies that reverse the inflammatory and immunogenic alterations that occur in infested fish.

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Although high diversity of parasitic ciliates has been reported in China, little is known about the species from high altitude areas, especially in Tibet. To investigate the species of parasitic ciliates in Tibet, a project was initiated in the Chabalang wetland in 2013. Two Trichodina species, namely, Trichodina sp. and T. reticulata Hirschmann & Partsch, 1955, were isolated from gills of an invasive fish, Micropercops swinhonis for the first time. In the present study, we provided the morphological, morphometrical, and molecular characterizations of the two species and conducted the phylogenetic analyses of mobilids based on the small subunit ribosomal RNA gene (SSU rDNA) sequences. Both morphological characters and morphometric data of the T. reticulata agreed well with previous studies. Although two partial SSU rDNA sequences were obtained in the present study, only the sequence of T. reticulata population in the present study was thought to be reliable. The other sequence may not belong to the other species. Thus, we regarded the other species isolated in the present study as Trichodina sp. to avoid the wrong or confused species identification. Morphologically, Trichodina sp. is distinguished mainly by its large body shape with a broad adhesive disk, robust and obliquely quadrilateral blades, and well-developed rays. T. reticulata is mainly characterized with the 8-12 spherical or elliptical granules in the central zone of adhesive disk. Phylogenetic analyses consistently showed the two ectoparasites clustered with freshwater species of the genus Trichodina within the order Mobilida. Our study extended the host range of T. reticulata and supplemented the molecular data. Also, results reveal that invasion of exotic fish may cause a potential threat to native fish by introducing or dispersing parasitic ciliates.

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Ciliates within the order Mobilida comprise a group of ectosymbionts that form ecologically and economically important symbioses with aquatic organisms. However, despite their significance, the factors driving mobilid diversification are poorly understood. To address this, we sought to examine the genetic diversity of two mobilid families, the trichodinids and urceolariids, over geographical areas and from different hosts. To this end we collected and provide morphological and molecular characterizations of three species of Trichodina: Trichodina domerguei and Trichodina tenuidens from three-spined sticklebacks, and Trichodina unionis from snails. We also sequenced the small subunit rRNA gene from Urceolaria korschelti collected from its type host at the type locality of Helgoland, Germany. The Trichodina from sticklebacks show intra-specific genetic variation that is not related to geographic distribution. Furthermore, we find evidence of fish to mollusc host transitions within the Trichodinidae and suggest that the most recent common ancestor of the Trichodina was a symbiont of vertebrates. Lastly we confirm that U. korschelti constitutes a cryptic species complex, which in turn justifies the description of Urceolaria parakorschelti sp. nov. These data suggest that not only host transitions, but allopatric and or genetically-driven sympatric speciation play a role in facilitating the diversification of mobilid ciliates.

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Trichodinids are ciliated protozoan parasites or symbionts of marine and freshwater organisms. During a parasitic ciliate survey in Hubei, China, two Trichodina species, T. acutaLom, 1961 and T. hyperparasitis Chen & Hsish, 1984, were isolated from body surface of loach Misgurnus anguillicaudatus and gills of yellow catfish Pelteobagrus fulvidraco, respectively. The two species were investigated morphologically, based on silver-strained specimens, and their small subunit ribosomal RNA gene (SSU rDNA) sequences were sequenced. Morphologically, T. acuta is distinguished mainly by the acute sickle-shaped blade with well-developed sharp blade apophysis, the irregular circular granule, and the robust ray. Trichodina hyperparasitis is mainly characterized by the broad blade and the inconspicuous round ray apophysis. Phylogenetic analyses revealed that the two Trichodina species investigated were nested within a clade including several freshwater Trichodina species. This indicates that the central granule is a useful taxonomic feature, but it may not be an important phylogenetic characteristic. Histopathological analysis revealed that T. hyperparasitis firmly colonized gills, which resulted in discrete hyperplasia and injuries of the gill filaments.

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The present study was aimed at screening for the presence of protozoan's among Cyprinid fishes collected from various fish ponds and farms in Jammu division of Jammu and Kashmir (J and K) state. Out of 75 fishes collected from local water bodies of Jammu division, only 35, (49.6 %) were infested with Trichodina. Trichodina infestations were studied in the period from November to February. In light infestation Trichodina was usually present on gills, fins and skin of apparently healthy fish. Clinical signs of Trichodiniasis appeared on fish with heavy infections and in presence of one or more stress factors including, rough handling during transportation from ponds, over crowdedness, malnutrition, high concentration of free ammonia and low oxygen concentration. Clinical signs of Trichodiniasis in fish such as sluggish movement, loss of appetite, black colouration, necrosis and ulcer on different parts of the body, detached scales and excessive accumulation of mucous in gill pouches were also observed.

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Water-soluble chlorophyll (chlorophyllin) was used in a phototoxic reaction against a number of fish ectoparasites such as Ichtyobodo, Dactylogyrus, Trichodina, and Argulus. Chlorophyllin is applied to the water at concentrations of several micrograms per milliliter for a predefined incubation time, and afterwards, the parasites are exposed to simulated solar radiation. Application in the dark caused only little damage to the parasites; likewise, light exposure without the addition of the photosensitizer was ineffective. In Ichthyobodo, 2 µg/mL proved sufficient with subsequent simulated solar radiation to almost quantitatively kill the parasites, while in Dactylogyrus, a concentration of about 6 µg/mL was necessary. The LD50 value for this parasite was 1.02 µg/mL. Trichodina could be almost completely eliminated at 2 µg/mL. Only in the parasitic crustacean Argulus, no killing could be achieved by a photodynamic reaction using chlorophyllin. Chlorophyllin is non-toxic, biodegradable, and can be produced at low cost. Therefore, we propose that chlorophyllin (or other photodynamic substances) are a possible effective countermeasure against several ectoparasites in ponds and aquaculture since chemical remedies are either forbidden and/or ineffective.

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Trichodina heterodentata Duncan, 1977 and Ichthyophthirius multifiliis Fouquet, 1876 obtained from gills during a parasitological survey conducted for the protozoan parasitic fauna of Schizothorax niger a snow trout in Dal Lake, Kashmir, India during the period October 2013 and March 2015. Thirty out of 180 fish were found infected with protozoan parasites. During the study of their pathogenecity the most common deteriorating signs observed in gill tissue were necrosis, hypertrophy, hyperplasia and fusion of secondary lamellae. Prevalence of infection was found to be 16.66%. This is the first record of the protozoan fauna of the schizothoracines from Kashmir valley, India.

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Trichodinids are ciliated protozoans that reversibly attach to the tegument of marine and freshwater host-organisms via an adhesive disc. In this study, we have used permeabilized cell models of Trichodina pediculus to examine the distribution of centrins, a Ca(2+)-binding protein associated with centrioles and/or contractile filamentous structures in a large number of protists. The previous finding that filamentous material of the adhesive disc comprised a 23-kDa centrin analog suggested that this protein might be a disc-specific isoform. This possibility was explored through immunolabeling methods using two distinct antibodies, anti-ecto-endoplasmic boundary (EEB) and anti-Hscen2 previously shown to react respectively with centrin-based filament networks and with centrioles. Immunofluorescence microscopy showed that anti-EEB reacts with filamentous material of the disc but not with basal bodies. Conversely, anti-Hscen2 cross-reacted with basal bodies but failed to label any type of structure occurring in the disc area. More detailed data on localization of this protein was obtained by immunoelectron microscopy showing gold particles deposits in the lumen of basal bodies. The different patterns revealed by this immunochemical approach suggest that the two protein antigens concerned by this study are distinct centrin isoforms that presumably perform organelle-specific function in the ciliate T. pediculus.

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