Clinical, histopathological and phylogenetic analysis of Myxobolus lentisturalis (Myxozoa: Myxobolidae) infecting the musculature of farmed population of goldfish (Carassius auratus) in Iran: 2021-2022.

There is a claimed increase in the global prevalence and incidence of emerging diseases observed in many organisms. Myxozoa represents an essential group of metazoan parasites that hold both economic and ecological significance. In the current study, 1% of the fish population at two commercial goldfish (Carassius auratus) farms in Tehran and Ghom province, Iran, developed cavitating muscular lesions resembling humps in February 2021 and January 2022. Fish displaying pathological abnormalities were transported to the Ornamental Fish Clinic and subjected to clinical examination. Light microscopy was subsequently used to examine wet smears of skin and gills, as well as whitish exudate. In addition, tissue homogenates were collected for more precise identification and molecular confirmation. The study discovered that individuals from the goldfish farms were infected with the pathogenic myxozoan Myxobolus lentisuturalis, which caused significant damage to the epaxial muscles. The spores collected from the humps had a lack of uniformity and were primarily ellipsoidal in shape. Histopathological analysis also revealed parasites in various stages of development, such as plasmodia and spores, as well as inflammatory cell infiltration (macrophage, giant cell and lymphoplasmacytic infiltration) between skeletal muscle fibers. Phylogenetic analysis of M. lentisuturalis was performed by using MEGA 11 and the maximum likelihood method. M. lentisuturalis is a myxozoan parasite that has been sparsely recorded and lacks widespread recognition. The current study is the first clinical, histopathological, and molecular characterization of M. lentisuturalis isolated from the skeletal musculature of goldfish (C. auratus) in Iran.

White spots amidst the gold: ultrastructural and histological aspects of the chronic inflammatory response of goldfish with ichthyophthiriasis.

Ichthyophthirius multifiliis, the causative agent of white spot disease, is a ciliated protozoan parasite that infects freshwater fish and induces high mortality. Outbreaks occur both in natural and production sites. The aim of the present study was to describe the lesions caused by chronic infection by I. multifiliis in goldfish (Carassius auratus) from an ornamental fish farm, highlighting important ultrastructural aspects of this protozoan. Damaged skin and gills, collected from fish with white or ulcerative skin lesions, were routinely processed for histological analysis and transmission electron microscopy. The parasitic forms present in the skin were associated with an inflammatory infiltrate consisting of macrophages, lymphocytes and other polymorphonuclear cells. The lesions associated with the presence of the parasite were organized in the form of granulomas, with macrophages in the layers closest to the parasites. A trophont-thickened membrane and induction of granulomatous inflammation were identified in this study as mechanisms for evasion of the immune response. We concluded that the presence of I. multifiliis trophonts resulted in the formation of granulomatous inflammation, whether associated or not with pathogen lysis, suggesting that the parasite can use an inflammatory response to evade the immune response.

Exploring the Efficacy of Pellitorine as an Antiparasitic Agent Against Argulus: Impacts on Antioxidant Levels and Immune Responses in Goldfish (Carassius auratus).

INTRODUCTION: Argulus spp. infestation is a significant challenge for aquaculture, currently, there are no approved medications available to efficiently manage this parasite. Consequently, mechanical removal of parasites using forceps and natural substances like herbs are being explored as alternative treatment methods. Pellitorine (PLE) is a naturally occurring compound found in several plant species. It is classified as an alkaloid and belongs to the class of compounds known as amides. MATERIALS AND METHODS: This study aimed to evaluate the effectiveness of PLE in preventing Argulus spp. infestations in goldfish (Carassius auratus) and to determine the optimal dosage of PLE for the detachment of Argulus spp. RESULTS: The findings of this study revealed that PLE enhanced the immune response of goldfish by promoting superoxide dismutase (SOD) and catalase (CAT) in Argulus-infected goldfish. Additionally, PLE induces reactive oxygen species (ROS) generation and cellular damage in the Argulus. PLE at a dosage of 5 mg/mL was able to detach 80% of the argulus from goldfish within 12 h. Therapeutic index was found to be 5.99, suggesting that PLE is the safest drug. CONCLUSIONS: Therefore, our findings suggest that PLE can be a suitable and effective treatment option for preventing Argulus infestations in goldfish. The results of this study can guide the use of PLE at an optimal dosage to control Argulus infestation in goldfish.

Myxobolus nekrasovae n. sp. (Cnidaria, Myxozoa) is a new species parasitizing the gills of the gibel carp, Carassius auratus gibelio.

A new Myxobolus species, Myxobolus nekrasovi n. sp., was found in the gill arch of the gibel carp Carassius auratus gibelio during investigation of fish myxosporean fauna of ponds of Lake Baikal basin. The parasites were studied on the basis of spore morphology, as well as with histological and molecular methods. Mature spores of M. nekrasovi n. sp. are ellipsoidal in frontal view and lemon-shaped in lateral view, measuring 13.84 ± 0.4 (12.2-15) µm in length, 9.73 ± 0.2 (8.5-10.7) µm in width, 6,75 ± 0.1 (6.0-7.6) µm in thickness. Polar capsules are unequal and pyriform, measuring: length 6.31 ± 0.1 (5.4-7.4), width 3.49 ± 0.04 (3.12-4) µm and length 2.88 ± 0.1 (2.1-3.5), width 1.4 ± 0.03 (1-1.6) µm. Phylogenetic analysis with the SSU rDNA gene shows Myxobolus nekrasovae n. sp. as a sister species of the subclade formed by Thellohanellus sinensis, Myxobolus acutus, M. zhaltsanovae that infect gibel carp Carassius auratus gibelio.

Efficacy of dietary live or heat-killed Bacillus subtilis in goldfish (Carassius auratus) infected with Ichthyophthirius multifiliis.

BACKGROUND: The beneficial effects of Bacillus subtilis on growth, immune response, and disease resistance against various diseases in different fish species have been proved. However, there are no data concerning this probiotic effect on skin mucosal immunity in fish infected with Ichthyophthirius multifiliis (Ich). Ich has a high mortality rate in both edible and ornamental fish and consequently is concerned with heavy economic losses. OBJECTIVES: Thus, we assessed the efficacy of live and heat-killed B. subtilis on skin immunity and histopathology in goldfish (Carassius auratus) infected with Ich. METHODS: Goldfish (144 fish, 2.38 g average weight) were stocked in nine glass tanks each in three replicates. Fish were fed 109 CFU g-1 live or heat-killed B. subtilis for 80 days. RESULTS: Probiotic administration in both viable and non-viable forms could enhance the growth performance in goldfish. Probiotic therapy also reduced the density of the parasite and histopathological level on skin and gill tissues of the treated fish. Real-time polymerase chain reaction analysis showed a higher expression of lysozyme and tumour necrosis factor-α in the treated groups compared to the control group. CONCLUSIONS: These data demonstrated the beneficial effect of B. subtilis as probiotic and paraprobiotic on growth performance and disease resistance to Ich infestation in goldfish.

Myxobolus lentisuturalis infection in a farmed population of goldfish Carassius auratus from the USA.

Myxobolus lentisuturalis is a myxozoan parasite of piscine muscle that has been described in goldfish Carassius auratus and Prussian carp Carassius gibelio. This report documents a naturally occurring infection of M. lentisuturalis in a population of farmed goldfish in the USA. Postmortem examination was performed on 4 affected goldfish. Gross findings included large cystic cavities along the dorsal midline filled with caseous exudate. Histopathology revealed myxozoan plasmodia and spores in the epaxial muscles with varying degrees of granulomatous and necrotizing myositis accompanied by lymphohistiocytic meningoencephalitis. Spore morphology and dimensions were consistent with M. lentisuturalis, as observed by light microscopy. PCR and sequence analysis of the small subunit ribosomal DNA of infected muscle samples from 2 goldfish confirmed the parasite to have 99-100% nucleotide identity to M. lentisuturalis sequences recovered from similar cases of this parasite infecting goldfish in China and Italy and Prussian carp in China. This is the first reported case of M. lentisuturalis in the USA and furthers the understanding of the pathogenicity of this under-described parasite.

In vitro and in vivo action of turmeric oil (Curcuma longa L.) against Argulus spp. in goldfish (Carassius auratus).

Background: Argulus is a common and widespread ectoparasite that causes major parasitic diseases and is a virus and bacteria carrier in the ornamental fish trade. Aim: The purpose of this study is to determine what types of phytochemicals are present in the essential oil extracted from turmeric rhizome (Curcuma longa L.) and to assess the efficacy of turmeric oil in eliminating Argulus spp. infestations in goldfish (Carassius auratus). Methods: The chemical composition and quantity of the major substances in essential oils from fresh turmeric rhizome were detected by gas chromatography/mass spectrometry (GC-MS). The antiparasitic effect of turmeric oils on Argulus spp. was tested at 12.5, 25, 50, 100, and 200 ppm and compared to 0.25 ppm Neguvon® (the positive control). The percentage of Argulus spp. killed, the percentage of the mean mortality rate, and the effectiveness of each test were evaluated. Results: By using GC-MS analysis, it was possible to identify the primary phytochemical component of turmeric essential oil as b-turmerone. The results obtained from the in vitro test indicated that there was a correlation between the concentration of turmeric essential oil and the average mortality rate of fish lice. The mean mortality of fish louse exposed to 200 ppm turmeric essential oil was higher than the mean mortality of fish louse exposed to Neguvon® (p < 0.05). In an in vivo study, the effectiveness of 12.5 ppm turmeric essential oil against parasites was 44.44%, 55.46%, and 62.83% at 24, 48, and 72 hours, respectively. Conclusion: In summary, the efficacy of turmeric essential oil against fish louse has been shown both in vitro and in vivo studies.

Description of Myxobolus hoabinhensis n. sp. (Myxosporea: Myxobolidae), infecting the trunk muscles of goldfish Carassius auratus (Linnaeus, 1758) (Cypriniformes: Cyprinidae) in northern Vietnam.

During a survey of myxosporean parasites of freshwater fishes in northern Vietnam, myxospores resembling those of the genus Myxobolus (Myxosporea: Myxobolidae) were found in the trunk muscle of 6 out of 35 specimens (17.14%) of wild goldfish Carassius auratus (Cypriniformes: Cyprinidae). The mature spores were 12.0 ± 0.4 (11.4 - 12.6) µm long, 8.5 ± 0.2 (7.9 - 9.0) µm wide and 6.1 ± 0.2 (5.8 - 6.3) µm thick, containing two pyriform-shaped polar capsules unequal in size. The larger polar capsule was 7.6 ± 0.3 (7.1 - 8.4) µm long and 3.5 ± 0.1 (3.3 - 3.8) µm wide, and the smaller polar capsule was 6.2 ± 0.3 (5.5 - 6.7) µm long and 2.9 ± 0.2 (2.6 - 3.4) µm wide. Each polar capsule contained a polar filament with 3-5 coils. A phylogenetic analysis based on the small subunit rDNA (SSU rDNA) sequence revealed that this Myxobolus species forms a distinct branch in the phylogenetic tree sister to Myxobolus artus and Myxobolus cyprini, with DNA sequence similarity at 97.6% to M. artus and 97.5% to M. cyprini. A combination of the morphological characteristics and molecular data suggest that this is an undescribed species, and we propose the name Myxobolus hoabinhensis n. sp.

Population growth of Gyrodactylus kobayashii in goldfish (Carassius auratus) associated with host density.

Host density is a key regulatory factor in parasite transmission. The goldfish (Carassius auratus)-Gyrodactylus kobayashii model was used to investigate effects of host density on population growth of gyrodactylids. A donor fish infected by five gravid gyrodactylids was mixed with 11 parasite-free goldfish at five host densities. There was a significant positive correlation between host density and mean abundance of G. kobayashii throughout the 58-day experiment. During early infection (days 15­24), mean abundance in medium high (0.5 fish L−1) and high host density groups (1 and 2 fish L−1) was significantly higher than that in the low host density groups (0.125 and 0.25 fish L−1). At high host density, prevalence increased more rapidly, and the peak prevalence was higher. Fitting of an exponential growth model showed that the population growth rate of the parasite increased with host density. A hypothesis was proposed that higher host density contributed to increased reinfection of detached gyrodactylids. A reinfection experiment was designed to test this hypothesis. Both mean abundance and prevalence at a host density of 1 fish L−1 were significantly higher than those at 0.25 fish L−1 on days 1 and 3, which suggested that more reinfections of G. kobyashii occurred at the higher host density. Density-dependent transmission during the early infection was an important contributor of population growth of G. kobayashii, as well as density-dependent reinfection of the detached gyrodactylids.

Ononin: A candidate anti-parasitic drug isolated from Spatholobi caulis against infections of Dactylogyrus intermedius (Monogenea).

Dactylogyrus is a common parasitic pathogen, which causes high mortality of fish when presents in large numbers, resulting in serious economic losses. Herbal medicines contain myriad of bioactive compounds is a valuable reserve for developing safe and effective anti-parasite drugs. Here, we conducted bioassay-guided fractionation to isolate and identify the anti-parasitic constituents from Spatholobi caulis. Among five extraction solvents (petroleum ether, chloroform, ethyl acetate, methanol and water), S. caulis methanolic extract had the highest parasiticide activity in Carassius auratus, and therefore subjected to further separation and purification using multiple chromatography methods. One compound exhibiting the strongest parasiticidal activity was obtained and identified as ononin by analyzing its spectral data (NMR and ESI-MS). The EC50 value of ononin against Dactylogyrus was 0.655 mg/L and showed 100% parasiticide activity with 3.0 mg/L. The 24, 48, 72, 96 h LC50 for goldfish were 4.691 (the 95% CI of 4.526-4.873) mg/L, 4.612 (4.441-4.800) mg/L, 4.472 (4.345-4.607) mg/L, 4.288 (4.155-4.428) mg/L, respectively. The present results discovered for the first time that ononin had potent parasiticidal activity and have the potential to be developed as new anti-parasitic drug for the control of Dactylogyrus.

Anthelmintic efficacy of natural saponins against Gyrodactylus kobayashii in goldfish (Carassius auratus) and their 3D-QSAR analysis.

Gyrodactylus spp. are common monogenean ectoparasites that may lead to significant fish mortality. To find effective anthelmintic agents with lower toxicity, a series of natural saponins were obtained and evaluated for their anthelmintic activity against Gyrodactylus kobayashii and acute toxicity to goldfish (Carassius auratus). Among all tested compounds, six compounds (1, 2, 3, 8, 10, and 13) shown higher anthelmintic activity and safety than widely used formaldehyde-based parasiticides, especially compound 1 having 100% anthelmintic efficacy against G. kobayashii at 0.3 mg/L and a therapeutic index of 16.6. Also, the three-dimensional quantitative structure-activity relationship (3D-QSAR) studies of these saponins have been performed to explore the structural features reasonable for the anthelmintic activity against G. kobayashii. These models demonstrated that the hydroxyl group at C-17 position and the sugar moieties at C-3 position, especially the hydroxyl groups of the sugar moieties, were critical to the anthelmintic activity. The QSAR studies could provide useful information for further rational design and optimization of novel saponins for the control of gyrodactylosis.

Innate cell-mediated cytotoxicity of CD8+ T cells against the protozoan parasite Ichthyophthirius multifiliis in the ginbuna crucian carp, Carassius auratus langsdorfii.

Cytotoxic T cells are known to have the ability to kill microbe-infected host cells, which makes them essential in the adaptive immunity processes of various vertebrates. In this study, we demonstrated innate cell-mediated cytotoxicity of CD8+ T cells against protozoan parasites found in the ginbuna crucian carp. When isolated effector cells such as CD8+, CD4+ (CD4-1+), or CD8- CD4- (double-negative, DN), from naïve ginbuna crucian carp were co-incubated with target parasites (Ichthyophthirius multifiliis), CD8+ cells from the kidney and gill showed the highest cytotoxic activity. On the other hand, DN cells, which include macrophages and CD4- CD8- lymphocytes, showed the lowest cytotoxic activity against I. multifiliis. Additionally, the cytotoxic activity of CD8+ cells was found to significantly decrease in the presence of a membrane separating the effector cells from I. multifiliis. Furthermore, the serine protease inhibitor 3,4-dichloroisocoumarin and perforin inhibitor concanamycin A significantly inhibited the cytotoxic activity of CD8+ cells. These results demonstrate that CD8+ T cells of ginbuna crucian carp can kill extracellular parasites in a contact-dependent manner via serine proteases and perforin. Therefore, we conclude that CD8+ T cells play an essential role in anti-parasite innate immunity of teleost fish.

Transcriptome analysis of goldfish (Carassius auratus) in response to Gyrodactylus kobayashii infection.

Gyrodactylid monogeneans are widespread parasites of teleost fishes, and infection with these parasites results in high host morbidity and mortality in aquaculture. To comprehensively elucidate the immune mechanisms against Gyrodactylus kobayashii, the transcriptome profiles of goldfish (Carassius auratus) skin after challenge with G. kobayashii were first investigated using next-generation sequencing. Approximately 21 million clean reads per library were obtained, and the average percentage of these clean reads mapped to the reference genome was 82.25%. A total of 556 differentially expressed genes (DEGs), including 344 upregulated and 212 downregulated genes, were identified, and 380 DEGs were successfully annotated and assigned to 95 signaling pathways in Kyoto Encyclopedia of Genes and Genomes (KEGG). In addition, 14 pathways associated with immune response were identified mainly including mTOR signaling pathway, cytokine-cytokine receptor interaction, intestinal immune network for IgA production, toll-like receptor signaling pathway, and phagosome. Twelve genes were selected and validated using qRT-PCR. A similar trend of these genes between RNA-Seq and qRT-PCR was observed, indicating that RNA-Seq data was reliable. Besides, the ALP activity and NO content in serum were significantly higher in the infected goldfish compared with the non-infected goldfish. In summary, this study provides better understandings of immune defense mechanisms of goldfish against G. kobayashii, which will support future molecular research on gyrodactylids and facilitate the prevention and treatment of gyrodactylosis in aquaculture.

Molecular and light microscopy evidence for the transfer of Myxobolus honghuensis from Carassius auratus gibelio broodfish to progeny.

Myxozoans usually have a complex life cycle involving indirect transmission between vertebrate and invertebrate hosts. The vertical transmission of these parasites in vertebrate hosts has not been documented so far. Here, we assessed whether the myxozoan parasite Myxobolus honghuensis is vertically transmitted in naturally infected allogynogenetic gibel carp Carassius auratus gibelio (Bloch). M. honghuensis infection of broodfish, fertilized eggs and laboratory-cultured progeny was monitored in 2018 and 2019. The presporogonic stage was microscopically observed in the pharynx of broodfish and their progeny. In situ hybridization confirmed the presence of M. honghuensis presporogonic stage in the pharynx of broodfish and progeny. Nested PCR results showed that M. honghuensis was present in tissues and eggs of broodfish, fertilized eggs and their corresponding progeny. The sequences obtained from broodfish and progeny showed 98.0-99.8% similarity with ITS-5.8S rDNA of M. honghuensis. This study provides molecular and light microscopy evidence for the transfer of M. honghuensis from broodfish to progeny via the eggs, but it is insufficient to assert that M. honghuensis can transmit vertically in naturally infected allogynogenetic gibel carp. This is the first record about vertical transfer of myxozoan in the vertebrate host.

The impacts of geographic and host species isolation on population divergence of Myxobolus lentisuturalis.

Samples of Myxobolus lentisuturalis were found in the gallbladder of Carassius auratus in Chongqing, China, without obvious disease symptoms, which were different from samples reported in Hubei, China, and Italy which were described as highly pathogenic muscle-infecting species. In order to improve our understanding of the relationships between these different samples, we analyzed geography, DNA sequence data, and site specificity. The results indicated that (1) the genetic relationship between Chongqing and Italy samples of M. lentisuturalis was much closer than relationship between each of them and the Hubei samples; (2) host species isolation was more important than the geographic isolation in divergence of M. lentisuturalis samples, and the species might be specialized among its different host species; and (3) geographic isolation and infection-site variation played a limited impact in genetic differentiation among different samples of M. lentisuturalis infecting the same host species.

Trypanosoma carassii infection in goldfish (Carassius auratus L.): changes in the expression of erythropoiesis and anemia regulatory genes.

Trypanosoma carassii is a flagellated bloodstream parasite of cyprinid fish with pathogenesis manifesting primarily as anemia in experimentally infected fish. This anemia is characterized by decreases in the number of circulating red blood cells (RBCs) during peak parasitemia. We examined changes in the key blood metrics and expression of genes known to be important in the regulation of erythropoiesis. Increasing parasitemia was strongly correlated with an overall decrease in the total number of circulating RBCs. Gene expression of key erythropoiesis regulators (EPO, EPOR, GATA1, Lmo2, and HIFα) and proinflammatory cytokines (IFNγ and TNFα) were measured and their expressions differed from those in fish made anemic by injections of phenylhydrazine (PHZ). Significant upregulation of pro-erythropoietic genes was observed in PHZ-induced anemia, but not during peak parasitic infection. Previously, we reported on functional characterization of goldfish erythropoietin (rgEPO) and its ability to induce survival and differentiation of erythroid progenitor cells in vitro. Treatment of goldfish during the infection with rgEPO reduced the severity of anemia but failed to fully prevent the onset of the anemic state in infected fish. Proinflammatory cytokines have been implicated in the suppression of erythropoiesis during trypanosomiasis, specifically the cytokines TNFα, IFNγ, and IL-1ß. Analysis of key proinflammatory cytokines revealed that mRNA levels of IFNγ and TNFα were upregulated in response to infection, but only TNFα increased in response to PHZ treatment. Synergistic activity of the proinflammatory cytokines may be required to sustain prolonged anemia. These findings provide insight into the relationship between T. carassii and host anemia and suggest that T. carassii may directly or indirectly suppress host erythropoiesis.

Parasite Dispersal From the Ornamental Goldfish Trade.

Goldfish, Carassius auratus Linnaeus, 1758, are immensely popular ornamental cyprinid fish, traded in more than 100 countries. For more than 500 years, human translocation has facilitated the spread of goldfish globally, which has enabled numerous and repeated introductions of parasite taxa that infect them. The parasite fauna assemblage of goldfish is generally well documented, but few studies provide evidence of parasite coinvasion following the release of goldfish. This review provides a comprehensive synopsis of parasites that infect goldfish in farmed, aquarium-held, native, and invasive populations globally and summarises evidence for the cointroduction and coinvasion of goldfish parasites. More than 113 species infect goldfish in their native range, of which 26 species have probably coinvaded with the international trade of goldfish. Of these, Schyzocotyle acheilognathi (Cestoda: Bothriocephalidae), Ichthyophthirius multifiliis (Ciliophora: Ichthyophthiriidae), Argulus japonicus (Crustacea: Argulidae), Lernaea cyprinacea (Crustacea: Ergasilidae), Dactylogyrus anchoratus, Dactylogyrus vastator and Dactylogyrus formosus (Monogenea: Dactylogyridae) are common to invasive goldfish populations in more than four countries and are considered a high risk of continued spread. Coinvasive parasites include species with direct and complex life cycles, which have successfully colonised new environments through utilisation of either new native hosts or suitable invasive hosts. Specifically, I. multifiliis, A. japonicus and L. cyprinacea can cause harm to farmed freshwater fish species and are important parasites to consider for biosecurity. These species may threaten other aquatic animal industries given their low host specificity and adaptable life histories. Future attention to biosecurity, management and border detection methods could limit the continued spread of exotic parasites from the ornamental trade of goldfish.

Identification of a new myxosporean parasite Thelohanellus indiana n. sp. (Myxosporea: Myxobolidae) isolated from three major organs of goldfish, Carassius auratus L. highlighted with its morphological and SSU rDNA sequence based molecular description.

Fish mortality and poor growth in surviving fish contribute substantial losses to the ornamental fish farms of India and revealed an infection of a new myxosporidian parasite Thelohanellus indiana n. sp. which has become one of the most important limiting factors for successful aquaculture management. The parasite infects Carassius auratus, an Indian goldfish, described on the basis of myxospores morphology and amplification of a part of 18 S rDNA gene. Three major attaching site of fish body have been explored for showing the location of attachment for the parasites. The whitish cysts of the parasites are about 2.5-3.5 mm contains large amount of lemon shaped mature myxospores measuring 12.1-15.2 (13.8) × 7.5-8.8 (8) µm. A single round or elliptical polar capsule located only at the anterior pole of the spore having 6.2-7.2 (6.8) × 3.3-4.7 (4.0) µm in diameter. The morphological characters have been assessed by both the light and scanning electron microscope. The most differentiating feature from closely related species was carried out by morpho-taxonomic affinities with previously described species which are tremendously supported by molecular taxonomy by partial sequencing of the 18 S rDNA gene resulted in a total of 2101 bp fragment of newly obtained SSU rRNA gene sequence of the new species which exhibit 79-91% homogeneity with other closely related species available in GenBank. The BLAST search of Thelohanellus sp. did not matches with any available sequences in GenBank and the phylogenetic analysis revealed that the novel species were sister to T. habibpuri and T. caudatus, in the Thelohanellus clade and form a closest neighboring branch as a subclade in phylogenetic tree from which the new Thelohanellus parasite is being placed. Both the branches are originating from monophyletic clade that are strongly supported by bootstrap values which indicate clearly about independent position of T. indiana n. sp.

Morphological, histological and molecular characterization of Myxobolus kingchowensis and Thelohanellus cf. sinensis infecting gibel carp Carassius auratus gibelio (Bloch, 1782).

A Myxobolus species and a Thelohanellus species infecting Carassius auratus gibelio (Bloch, 1782) were redescribed by their morphological, histological and molecular characterization. In the present study, the Myxobolus species infecting the muscle was identified as Myxobolus kingchowensis Chen et Ma, 1998 by the morphological and molecular data. Histologically, mature spores of M. kingchowensis were observed in the intercellular and connective tissue of muscle, though the plasmodia were not found. In addition, scattered spores also occurred in the intercellular of haematopoietic cells, intraepithelial of the renal tubules and interior of the melano-macrophage centres. Phylogenetic analysis showed that M. kingchowensis clustered in the clade of muscle-infecting Myxobolus species, further supporting muscle as the infection site of M. kingchowensis. The present Thelohanellus species infecting the gills was identified conspecific as Thelohanellus sinensis reported in Sun (2006) (mark it as T. sinensis-Sun)based on spore morphology, biological traits (host specificity and organ specificity), and molecular data. However, compared with the original description of T. sinensis Chen et Hsieh, 1960, the present Thelohanellus species and T. sinensis-Sun both infecting the gills of gibel carp are distinguishable from the original description in the host and infection site, which made the validity of T. sinensis-Sun dubious. Due to the absence of molecular data in the original description of T. sinensis, we suggest marking the present species and T. sinensis-Sun as T. cf. sinensis to avoid the confusion until T. sinensis is obtained from the type host and type infection site.

Composition and Diversity of Communities of Dactylogyrus spp. In Wild and Farmed Goldfish Carassius auratus.

Species composition and diversity of dactylogyrids were compared on gills of wild and cultured goldfish (silver crucian carp) Carassius auratus from 3 naturally populated lakes and 3 stocked aquaculture ponds in the Hubei province of China to examine the differences in the gill parasite community between these natural and farmed waters. Of the 7 Dactylogyrus spp. detected, all were found in lakes and 5 in ponds, with Dactylogyrus inexpectatus and Dactylogyrus anchoratus being absent from ponds. No significant correlation was found between the species richness and habitat area or host size, nor was there a significant difference in mean species richness between lakes (0.41-0.65) and ponds (0.30-0.76). Brillouin's diversity in lakes (0.049-0.067) was higher than that in ponds (0.024-0.046), but not significantly so. Although the diversity of parasite communities was higher in wild goldfish, higher mean abundance of some Dactylogyrus spp. was found in cultured goldfish. Based on Bray-Curtis similarity, it was difficult to differentiate parasite communities in lakes from those in ponds at the infracommunity level, whereas the 3 lakes and Guanqiao pond differed markedly from the remaining 2 ponds at the component community level. Although infracommunities differed among waterbodies, no effects of fish length or waterbody type were found on infracommunity or component community structure. Together, these results suggest that abundance and species richness of Dactylogyrus spp. on goldfish in lakes and farm ponds are influenced by habitat-specific environmental factors.