Gibel Carp Carassius Auratus Gibelio Research Articles

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.

Identification and analysis of differentially expressed microRNAs in gibel carp Carassius auratus gibelio responding to polyinosinic-polycytidylic acid (poly I:C) stimulation

Polyinosinic-polycytidylic acid (poly I:C) is a synthesized analogue of viral double-strand RNA and considered as a potential immunostimulant in aquaculture. MicroRNAs (miRNAs) have been reported to play important roles in the development of the immune system and in regulation of host antiviral responses. In our earlier study, it was found that poly I:C pre-treatment could stimulate the resistance against cyprinid herpesvirus 2 (CyHV-2) infection and enhance the antiviral immune response in gibel carp. To understand the role of miRNAs in regulating the host response to poly I:C treatment, we investigated the expression profiles of miRNAs in the head kidney of poly I:C-treated gibel carp with small RNA sequencing technology. When compared with the untreated group, a total of 24 differentially expressed miRNAs were identified in the poly I:C-stimulated fish, among which, 7 and 17 miRNAs were upregulated and downregulated, respectively. Analysis of target genes of these differentially expressed miRNAs found that most targeted mRNAs were involved in catalytic activity, peptidase activity and endopeptidase activity, and were enriched in the metabolic, protein processing in endoplasmic reticulum and oxidative phosphorylation signaling pathways, suggesting that poly I:C could alter the expression of metabolism-related miRNAs in the kidney of gibel carp. Besides, it was noted that some immune-related miRNAs, including inflammation-related miRNAs (miR-192 and miR-731) and interferon-related miRNAs (miR-194a and miR-122), were downregulated after poly I:C treatment. In summary, it was found that poly I:C could regulate the cellular levels of specific miRNAs involved in metabolism and immune responses in the head kidney of gibel carp, which may increase the capacity of the immune cells to fight against pathogens infection.

The melanocortin-4 receptor (MC4R) gene in the gibel carp Carassius auratus gibelio: cloning, tissue distribution, and fasting effects

The melanocortin-4 receptor (MC4R) gene in the gibel carp Carassius auratus gibelio: cloning, tissue distribution, and fasting effects

Does the Anionic Surfactant Isopropylamine Dodecylbenzene Sulfonate Induce Hepatic Oxidative Stress and Impairment in Carassius auratus gibelio?

Isopropylamine dodecylbenzene sulfonate (IDS) is a new kind of anionic surfactant (ANS). To preliminarily evaluate the aquatic toxicity of IDS, this study took gibel carp (Carassius auratus gibelio) as the research object. The well-acclimatized fish were divided into six groups and exposed to 0mg/L, 0.5mg/L, 1.0mg/L, 2.0mg/L, 4.0mg/L, or 8.0mg/L of IDS for 7, 14, 21 or 28 days. Our results showed that the superoxide dismutase (SOD) activity and malondialdehyde (MDA) content of the liver were unaffected by IDS exposure, while glutathione peroxidase (GSH-Px) activity was significantly inhibited. Hepatic tissue exhibited pathological damage, characterized by nuclear migration and dissolution and cell boundary blurring. The results suggest that IDS does not cause oxidative stress in the liver, but cause hepatic histopathological damage. GSH-Px can be considered as a biomarker of IDS exposure in gibel carp.

Assessment of aquatic genotoxicity of the Lake Sevan basin, Armenia using natural bioindicators

Water pollution can cause genomic instability, thereby threatening aquatic wildlife and human health․ For a correct assessment of the impact of the mixture of pollutants, chemical analysis of environmental contaminants should be simultaneously applied with genotoxicity assays. The сomet assay is a sensitive, versatile and extensively used method for genotoxicity assessment in a water environment and was successfully applied in aquatic wild species, including fish and crayfish. Earlier we investigated the genotoxicity of waters of the Lake Sevan basin (Simonyan et al. 2016, Simonyan et al. 2019)․ Lake Sevan is the largest lake of the Caucasus Region, situated in the Republic of Armenia, in the Gegharkunik Province. Water pollution in the lake is a result of both anthropogenic and natural pressures. A combination of comet assay (Tice et al. 2000) and chemical analysis was applied for evaluation of water genotoxicity using gibel carp Carassius auratus gibelio and crayfish Astacus leptodactylus as sentinel organisms. Fish and water samples were collected from the south–southwest of the village Shorzha (40028′33″N and 45014′22″E), Peninsula of Lake Sevan (40033′46″N and 45001′37″E), the estuaries of the Rivers Gavaraget (40025′12″N and 45009′53″E) and Dzknaget (40036′55″N and 44058′13″E). Сrayfish and water samples were collected from the estuary of the River Masrik (40013′25″N and 45038′21″E) and near the villages of Artanish (40027′19″N and 45025′12″E) and Tsapatakh (40024′34″N and 45028′22″E). According to the chemical analysis of waters, the south–southwest of the village Shorzha and the estuary of the River Masrik were chosen as reference sites․ The content of contaminants in water was analyzed according to the standard methods in the Environmental Impact Monitoring Center, Ministry of Environment of the Republic of Armenia. The obtained results showed that the levels of DNA damage were significantly higher in both species in polluted areas than in reference sites. A significant positive correlation between DNA damage in C. auratus gibelio and contents of Al, Fe, Cu and Mn in water was shown. DNA damage in A. leptodactylus correlated with Al, Fe, Cu and Mo. C. auratus gibelio and A. leptodactylus were found to be sensitive mostly to the same pollutants, which indicate that these metals may be of primary concern as contaminants of the studied aquatic environment. Thus, the current study indicates that C. auratus gibelio and A. leptodactylus are sensitive bioindicators for monitoring of water pollution in the Lake Sevan basin.

Development and evaluation of a convenient immunochromatographic strip test for rapid detection of cyprinid herpesvirus 2 (CyHV-2).

Cyprinid herpesvirus 2 (CyHV-2) has become a serious threat to the gibel carp Carassius auratus gibelio industry and has led to enormous losses worldwide. We have therefore developed an immunochromatographic strip (ICS) to enable rapid on-site detection of CyHV-2 by aquaculture facility staff. The ICS employs 2 monoclonal antibodies (MAbs 2C3-1E6 and 3H2-1G5) against the ORF25 protein, a CyHV-2 membrane protein, as the capture and detection antibodies, respectively. Indirect immunofluorescence assay (IIFA) and Western blotting of CyHV-2-infected fathead minnow cells indicated that the 2 MAbs could specifically bind CyHV-2 by recognizing ORF25 antigen. Sandwich ELISA showed that the detection limit of ORF25 protein halved when MAb 2C3-1E6 served as the capture antibody compared to MAb 3H2-1G5. The test for detecting purified CyHV-2 using the ICS could be completed in 10 min and the sensitivity was 1 µg ml-1. Sensitivity of the ICS remained stable following storage at 4, 25 and 37°C for 6 mo. Tissue homogenate from gibel carp with and without obvious gill hemorrhages was subjected to CyHV-2 detection using the ICS: the results were in good accordance with conventional PCR. Our ICS does not require highly trained technicians or specialized equipment, making it suitable for rapid diagnosis of CyHV-2 infection both in the laboratory and in the field.

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.

Function characterization and expression regulation of two different-sized 3’ untranslated region-containing interferon genes from clone F of gibel carp Carassius auratus gibelio

Fish interferon (IFN)-mediated antiviral innate immunity is the first line of defense against virus invasion. In the present study, we identify two fish IFN genes (here tentatively named IFNa and IFNc) with different-sized 3′ UTRs from clone F strain of gibel carp Carassius auratus gibelio. Carp IFNa has a relatively short 3′UTR without AU-rich elements (AREs) but IFNc has a long one with 9 AREs. Functionally, carp IFNa and IFNc display significantly antiviral potential to viral infection, likely through induction of downstream IFN-stimulated genes (ISGs). Both carp IFN genes are induced by viral infection, poly(I:C) treatment and IRF3/7, which are ascribed to the IFN-sensitive response elements (ISRE) within their promoters. Carp IFN genes are also induced by each other and by themselves, indicating existence of a positive feedback loop in fish IFN-mediated antiviral immune response. Comparative analyses of 3′UTR-mediated expression regulation at mRNA and protein levels show that the ARE-containing 3′UTR of carp IFNc rather than the short 3′UTR of carp IFNa promotes mRNA decay but instead results in high-level protein expression, indicating that 3′UTR of fish IFN mRNAs might be a potential factor for regulation of IFN-mediated antiviral immune response. Considering a fact that a given protein function is largely related to its protein level, these results suggest that both promoter and 3′UTR contribute to the transcription and translation of fish IFN genes, thus shaping their eventually antiviral potential.

Effects of a highly purified fucoidan fromUndaria pinnatifidaon growth performance and intestine health status of gibel carpCarassius auratus gibelio

Effects of a highly purified fucoidan from<i>Undaria pinnatifida</i>on growth performance and intestine health status of gibel carp<i>Carassius auratus gibelio</i>

Indigenous AHL‐degrading bacteriumBacillus firmussw40 affects virulence of pathogenicAeromonas hydrophilaand disease resistance of gibel carp

Interrupting quorum sensing represents a novel anti‐infective strategy to combat bacterial pathogen, and biodegradation of quorum sensing signal AHLs has been proved to be an efficient way to control pathogenic Gram‐negative bacteria in aquaculture. In this study, the effect of Bacillus firmus sw40 as efficient AHL‐degrading strain on virulence of fish pathogen Aeromonas hydrophila and disease resistance of gibel carp Carassius auratus gibelio was investigated. The results demonstrated that in vitro the B. firmus sw40 extracellular production (ECP) was able to significantly decrease protease production, haemolytic activity and biofilm formation in A. hydrophila. Dietary administration of B. firmus sw40 (10⁹ CFU/g) for 4 weeks significantly reduced the inflammatory cytokines TNF‐1a, TNF‐2a and IFN‐γ genes expression, antioxidant parameter MDA and GSH levels in serum and increased antioxidant enzyme SOD activity. Besides, B. firmus sw40 could significantly increase the survival of gibel carp with pathogenic A. hydrophila infection.

Difference in biochemical markers in the gibel carp (Carassius auratus gibelio) upstream and downstream of the hydropower plant

However the physiological stress in aquatic organisms associated with hydropower plants (HPP) ecosystems has been previously investigated, no studies have so far assessed it on biochemical level. Therefore this study evaluated an oxidative stress and toxicity in the gibel carp Carassius auratus gibelio associated with a small-scale HPP in the West Ukraine. A battery of liver, brain and blood markers was evaluated individuals inhabiting upstream and downstream of the dam of the small-scale Kasperivtci HPP (KHPP; an installed capacity of 7.5 MW), and from a reference site. Number of alterations were noted in fish from the KHPP impoundment facility including signs of oxidative stress (a decrease in superoxide dismutase (SOD) activity and an increase in protein carbonyls) and cytotoxicity (an increase in micronucleated erythrocytes and caspase-3 activity). No changes in DNA fragmentation in hepatocytes or brain cholinesterase activity were detected. As demonstrated by the integral stress index, fish associated with downstream of the dam revealed the greatest alterations reflected by the combined oppression of antioxidant system (SOD, catalase) and pro-oxidants (thiobarbituric acid reactive substances and oxyradicals), low concentration of metallothioneins, but high cathepsin D activity (as markers of lysosomal dysfunction and autophagy) and increased vitellogenin concentration in males (indicating an endocrine disruption). The study highlights that fish inhabiting ecosystems associated with HPP, particularly downstream of the dam, may face additional stresses with long-term effects yet to be evaluated.

Identification, expression profiles and antiviral activities of a type I IFN from gibel carp Carassius auratus gibelio

Type I interferons, as a class of multipotent cytokines, play a key role in host antiviral immune responses. In this study, a type I IFN coding gene of gibel carp, Carassius auratus gibelio, CagIFNa was cloned and sequenced. The full-length cDNA sequence of CagIFNa consists of 724 nucleotides that encode a predicted protein of 183 amino acids. CagIFNa has two highly conserved cysteine residues in the deduced protein, which is mostly conserved in the fish group I type I IFNs. CagIFNa was identified as a member of the IFNa subgroup of group I type I IFNs by phylogenetic analysis. CagIFNa transcripts were detected in all investigated tissues with higher levels in the liver, intestine, spleen and head kidney of gibel carp. Following injection with Cyprinid herpesvirus 2 (CyHV-2), CagIFNa gene expression was significantly inhibited in the spleen but delayed and then increased in head kidneys. Similarly, while CagIFNa expression was rapidly induced in gibel carp brain (GiCB) cells by poly I:C stimulation and its high induction level was delayed following CyHV-2 infection. CagIFNa overexpression in GiCB cells drastically reduced virus CPE and titer. Furthermore, several genes associated with type I IFN signaling pathway including IRF3, IRF7, IRF9, STAT1, Mx1 and PKR were induced in GiCB cells overexpressing CagIFNa upon CyHV-2 infection. These results show that CagIFNa plays a role in antiviral immune system in gibel carp.

Effects of genetically modified and non‐genetically modified soybeans with different heat treatments on growth and health of Cyprinidae species with different feeding habits

An 8-week growth trial was conducted to investigate the effects of non-genetically modified (nGM) soybean (Youchun 06-1) and genetically modified (GM) soybean (Roundup Ready®) with and without a heat treatment on the growth and health of three Cyprinidae species with different feeding habits (grass carp Ctenopharyngodon idellus, gibel carp Carassius auratus gibelio, and black carp Mylopharyngodon piceus; body weight: 283.0 ± 2.0 g, 60.5 ± 0.7 g, and 261.4 ± 3.1 g). Five diets (FM, nGMS, hnGMS, GMS, and hGMS) were made with fishmeal and full-fat soybean (Youchun 06-1, heat-treated Youchun 06-1, Roundup Ready®, and heat-treated Roundup Ready®). The FM diet contained fishmeal as the sole protein source. The experimental diets contained full-fat soybean meal as 60% of dietary protein. Both temperature (60, 80, 100, and 120°C) and duration (1 and 2 hr) of heating influenced trypsin inhibitor activity (TIA) and protein solubility of nGM soybean and GM soybean. The TIA in GM soybean was higher than that in nGM soybean. After heating at 120°C for 2 hr, each amino acid of soybean treatment showed a slight decline. Neither source of soybean nor heat treatment affected the growth performance, feed utilization, chemical composition of the whole fish body and muscle, and lipase or amylase activity of the three Cyprinidae species (p > 0.05). Heat treatment of soybean meal slightly increased the plasma antioxidant capacity of the three fish and plasma cholecystokinin of black carp and grass carp. The unheated soybean treatment adversely affected the height and density of black carp intestinal villi, and all the soybean treatments caused disruption of the grass carp intestinal epithelium.

Production performance of largemouth bass Micropterus salmoides and water quality variation in monoculture, polyculture and integrated culture

A 74‐day experiment was conducted to evaluate the production performance and water quality variation in three types of farming system for largemouth bass Micropterus salmoides. The tested aquaculture models included monoculture of largemouth bass (MC), polyculture of largemouth bass, gibel carp Carassius auratus gibelio and silver carp Hypophthalmichthys molitrix (PC), and integrated culture of largemouth bass, gibel carp, silver carp and freshwater pearl mussel Hyriopsis cumingii (IC). The ratio of largemouth bass, gibel carp and silver carp was 30:2:1 in the PC model, and the ratio of largemouth bass, gibel carp, silver carp and mussel was 30:2:1:5 in the IC model. The largemouth bass were fed with formulated feed twice daily. No significant differences were found in weight gain and yield of largemouth bass, total fish yield, nitrogen (N) and phosphorus (P) utilization efficiencies, N and P wastes, pH, nitrite, nitrate, reactive phosphate, total nitrogen, total phosphorus, total organic carbon, chemical oxygen demand, 5‐day biochemical oxygen demand, chlorophyll a, primary productivity among the MC, PC and IC models. The ammonia was lower, while the dissolved oxygen was higher in the PC tanks than in the MC tanks. These results suggest that the environment situation was better in the PC tanks relative to that in the MC tanks. The present study reveals that the PC model should be a way to optimize the aquaculture model for commercial largemouth bass farming.

Effects of high ammonia concentrations on three cyprinid fish: Acute and whole-ecosystem chronic tests

A number of studies have revealed ammonia to be toxic to aquatic organisms; however, little is known about its effects under natural conditions. To elucidate the role of ammonia, we conducted 96-h acute toxicity tests as well as a whole-ecosystem chronic toxicity test for one year in ten 600-m2 ponds. Three common cyprinids, silver carp Hypophthalmichthys molitrix Val. (H.m.), bighead carp Aristichthys nobilis Richardson (A.n.), and gibel carp Carassius auratus gibelio Bloch (C.g.), were used as test organisms. The 96-h LC50 values of un-ionized ammonia (NH3) for H.m., A.n., and C.g. were 0.35, 0.33, and 0.73mgL−1, respectively. In the ponds, annual mean NH3 ranged between 0.01 and 0.54mgL−1, with 4 ponds having a NH3 higher than the LC50 of A.n. (lowest LC50 in this study). No fish were found dead in the high-nitrogen ponds, but marked histological changes were found in livers and gills. Despite these changes, the specific growth rate of H.m. and A.n. increased significantly with NH3. Our pond results suggest that fish might be more tolerant to high ammonia concentrations in natural aquatic ecosystems than under laboratory conditions. Our finding from field experiments thus suggests that the existing regulatory limits for reactive nitrogen (NH3) established from lab toxicity tests might be somewhat too high at the ecosystem conditions. Field-scale chronic toxicity tests covering full life histories of fish and other aquatic organisms are therefore encouraged in order to optimize determination of the effects of ammonia in natural environments.

Development of a multiplex PCR method for the simultaneous detection of four myxosporeans infecting gibel carp Carassius auratus gibelio.

Gibel carp Carassius auratus gibelio (Bloch), a commercially important freshwater-cultured fish in China, is threatened by myxosporeans, particularly Thelohanellus wuhanensis, Myxobolus honghuensis, M. wulii and M. turpisrotundus. Here, we developed a multiplex PCR assay for simultaneous detection of these 4 myxosporeans. The specific primers for each species were designed based on the 28S rDNA gene of T. wuhanensis, the ITS-5.8S rDNA of M. honghuensis and M. wulii, and the 18S rDNA gene of M. turpisrotundus. Specificity testing confirmed that the 4 primer sets have no cross-reactivity with other related myxosporean species tested. Detection limits of the multiplex PCR assay were 0.2, 0.3, 3.1 and 3.8 spores for T. wuhanensis, M. honghuensis, M. wulii and M. turpisrotundus, respectively. Following screening of 104 field samples, the analytical sensitivity of the present multiplex PCR assay was found to be similar to the sensitivity obtained by the singleplex PCR assays and was higher than that of microscopic examination. Moreover, Kappa analysis showed a strong agreement between the results of the singleplex and multiplex PCR assays, indicating that the developed multiplex PCR assay was an efficient approach for the diagnosis of the 4 myxosporeans infecting gibel carp.

Histopathological and ultrastructural studies of Myxobolus turpisrotundus from allogynogenetic gibel carp Carassius auratus gibelio in China.

During an ongoing systematic survey on species diversity of myxozoans parasitising allogynogenetic gibel carp Carassius auratus gibelio (Bloch) in China, plasmodia were detected in the fins, lip, jaw, gill chamber, gill arches, operculum and oral cavity of infected fish. Combining the morphological and molecular data, the present species was identified as Myxobolus turpisrotundus Zhang, Wang, Li et Gong, 2010. Histopathological examination revealed that despite infecting different organs, M. turpisrotundus always occurred in dermis, demonstrating its affinity to this tissue. Histopathological effect of M. turpisrotundus on the host is relatively mild except parasites in the gill arches producing compression of the adipose tissue and heavy adductor muscles deformation with lymphohistiocytic infiltrates. In addition, the plasmodia in different sites were with the same complex structure arrangement: cup-like cells with unknown derivation, a thin collagenous fibril layer, areolar connective tissue, basement membrane and host epithelial cell. Ultrastructural analysis showed that the parasite has monosporic pansporoblast and sporogenesis followed the usual pattern of most of the myxosporeans.

Development of monoclonal antibodies against polar filaments and spore valves of Myxobolus honghuensis (Myxosporea: Bivalvulida).

Myxobolus honghuensis infects the pharynx of allogynogenetic gibel carp Carassius auratus gibelio (Bloch) and can cause high mortality. Only morphology-based diagnostic methods are currently available for clinical samples, but these methods are laborious and have low efficiency of detection. To overcome this problem, we designed a more sensitive diagnostic method. Two monoclonal antibodies (MAbs 1C7 and 3B7) were prepared by immunizing mice with soluble protein from sonicated M. honghuensis spores. Immunofluorescence analysis revealed that MAb 1C7 specifically reacts with polar filaments from spores, whereas MAb 3B7 identified protein localized on the spore valves. The isotypes of MAb 1C7 and MAb 3B7 were IgM and IgG1, respectively. Results of Western blot analysis revealed that MAb 1C7 recognized 2 prominent protein bands with molecular weights of 130 and 180 kDa, while MAb 3B7 recognized a protein band of 28 kDa. Thus, in this study we have developed 2 MAbs that have the potential for efficient detection of M. honghuensis. Moreover, identification of MAb 1C7 and MAb 3B7 allows for further studies of the functions and biochemical composition of polar filament and spore surface antigens.

Myxozoans and Our Dinner Table: Pathogenicity Studies of Myxobolus honghuensis (Myxosporea: Bivalvulida) Using a Suckling Mice Model.

Myxobolus honghuensis parasitizes the pharynx of the allogynogenetic gibel carp Carassius auratus gibelio (Bloch), an economically important food fish with large consumption rates in China. The fact that it is hard to detect the cysts formed by M. honghuensis without euthanizing the fish and the spores can reach the consumers has prompted us to investigate the effect of M. honghuensis on conventional, immunocompetent BALB/c suckling mice, in order to consider the possible pathological effects in general immunocompetent consumers by the ingestion of Myxobolus-infected fish. BALB/c suckling mice were inoculated with serial dilutions of the spore suspension and then hematological and histological studies were performed. Tests of suckling mice showed no statistically difference of fluid accumulation ratio between mice inoculated with 1.65 × 10(6) M. honghuensis spores and negative control after 8-h incubation. Spores did not disrupt intestinal histology, and no abnormal bowel movements were observed within 20 h postinoculation. No anomalous hematology parameters were documented. These results, taken together, reveal that M. honghuensis spores do not cause pathology (diarrhea and elevated fluid accumulation) in BALB/c suckling mice. However, an extrapolation is not currently feasible because of lack of a highly quantifiable risk extrapolation model, inaccessibility to data for the effect of M. honghuensis on humans, and lack of consideration of the immunosuppressed population. Our study extends the results of the effect of myxozoan on suckling mice and serves as baseline for future experiment using the same model in evaluating the myxozoan-caused foodborne disease.

Morphological and molecular characterization of actinosporeans infecting oligochaete Branchiura sowerbyi from Chinese carp ponds.

We surveyed the actinosporean stages of fish myxosporeans at fish farms in Jiangsu Province, China, from 2011 to 2014. During the surveys, we identified 7 actinosporean types from 4 collective groups: echinactinomyxon (1 type), triactinomyxon (1 type), aurantiactinomyxon (1 type), and neoactinomyxum (4 types), released by the oligochaete Branchiura sowerbyi. The morphological characteristics and DNA sequences of these types are described here. Based on 18S rDNA sequence analysis, the actinosporean of echinactinomyxon type CZ with 4 branches at the end of the caudal processes was identified as Myxobolus wulii, and the neoactinomyxum type JD was identified as Thelohanellus wangi Yuan, Xi, Wang, Xie, Zhang, 2015 (JX458816), a recently nominated species from the gills of allogynogenetic gibel carp Carassius auratus gibelio. In addition, actinosporeans of aurantiactinomyxon type JD, neoactinomyxum type CZ-1, neoactinomyxum type CZ-2, and neoactinomyxum type CZ-3 showed high genetic similarity to T. wuhanensis (96.3-96.5%), T. nikolskii (98.0-99.1%), T. wuhanensis (97.8-98.9%), and T. hovorkai (98.7-98.9%), respectively. Phylogenetic analyses showed that these actinosporeans were robustly clustered in the Thelohanellus spp. clade.