Predation of Cyclopoid Copepods on the Theronts of Ichthyophthirius multifiliis: Shedding Light on Biocontrol of White Spot Disease.

White spot disease, caused by the parasitic ciliate Ichthyophthirius multifiliis, is a significant threat to the freshwater fish farming industry worldwide, resulting in massive mortality and economic losses. Eliminating the free-swimming theronts from the culture environment is considered crucial for the control of I. multifiliis infection. It is well-documented that planktonic ciliates are valuable food resources for macro-zooplankton in aquatic ecosystems. In this study, we developed a fluorescence labeling method for alive theronts and found that cyclopoid copepods Thermocyclops taihokuensis, Mesocyclops spp., Macrocyclops sp., and Paracyclopina sp. present predation on the theronts in co-culture experiments. Laboratory challenge tests further confirmed that the presence of zooplankton in the culture water body significantly reduced the infection of I. multifiliis in goldfish (p < 0.01). Results from this study revealed that cyclopoid copepods have the potential to be used as biological control agents against white spot disease in aquaculture.

Characterization of the complete mitochondrial genome of Nippotaenia mogurndae Yamaguti and Miyata, 1940 (Cestoda: Nippotaeniidae).

In this study, we report the first complete mitochondrial genome of the tapeworm Nippotaenia mogurndae in the order Nippotaeniidea Yamaguti, 1939. This mitogenome, which is 14,307 base pairs (bp) long with an A + T content of 72.2%, consists of 12 protein-coding genes, 22 transfer RNA (tRNA) genes, two rRNA genes, and two non-coding regions. Most tRNAs have a conventional cloverleaf structure, but trnS1 and trnR lack dihydrouridine arms of tRNA. The two largest non-coding regions, NCR1 (220 bp) and NCR2 (817 bp), are located between trnY and trnS2 and between nad5 and trnG, respectively. Phylogenetic analyses of mitogenomic data indicate that N. mogurndae is closely related to tapeworms in the order Cyclophyllidea.

The erection of a new genus, Neotegorhynchus n. g. (Palaeacanthocephala, Illiosentidae), with a redescription of Neotegorhynchus cyprini n. comb. from Cyprinus carpio from the Yangtze River basin, China.

The present paper summarizes acanthocephalan parasites of the family Illiosentidae Golvan, 1960, collected from the intestine of common carp (Cyprinus carpio L.) from Taihu Lake in Wuxi, Jiangsu Province, China. A total of 21 acanthocephalan specimens were found in 6 carps (prevalence of 35%). All the studied specimens were assigned to the family Illiosentidae based on the family-specific morphology of the worms and the presence of 8 cement glands in the males. However, the specimens recently found in common carp differed from all 14 extant genera of the family Illiosentidae in the structure of the reproductive system of both sexes, i.e. i) a vagina lacking a muscular sphincter; ii) the presence of a terminally pointed protruding tail end in the form of a dome with a muscular base; iii) the female genital muscles are fan-shaped cells, each of which has a nucleus and is not attached to the anterior wall of the body; iv) the muscular lip of the bursa of males repeats the curved shape of the posterior end of females, which en copula allows the lip of the bursa to seal to the posterior end of the female. Morphologically Neotegorhynchus n. g. is closest to the genus Tegorhynchus, but differs from it, in addition above features in i) the terminal position of the genital pores of females without a hollow genital vestibule and without a transverse cleft connected to the dorsal terminal genital pore, as in Tegorhynchus brevis; ii) cerebral ganglion at the border of the anterior and middle third of the proboscis vessel; iii) spherical cement reservoir. Molecular studies confirmed Neotegorhynchus n. g. as belonging to the family Illiosentidae, showing less than 98.9% sequence similarity in SSU rDNA and 81.8% in COI with the genus Dentitruncus. Therefore, a new acanthocephalan genus, Neotegorhynchus n. g., is erected, and Neotegorhynchus cyprini n. comb. is designated as the type species and its neotype.

Molecular cloning and functional characterization of the hypoxia-inducible factor-1α in bighead carp (Aristichthys nobilis).

HIF-l is the earliest documented and most widely studied hypoxia-inducible factor (HIF) and plays a key role in the cell hypoxia signal transduction pathway. Particularly, the HIF-1α protein is sensitive to oxygen and plays a critical role in hypoxia regulation. This study is the first to report on the molecular cloning and characterization of HIF-1α in bighead carp (Aristichthys nobilis; anHIF-1α). The full-length cDNA of anHIF-1α was 2361 bp, and encodes an estimated 674 amino acids with a predicted molecular mass of 76.10 kDa and a theoretical isoelectric point of 7.72. Moreover, the conserved basic Helix-Loop-Helix domain along with two Per-ARNT-Sim domains (A/B), and C-TAD were identified in this protein. Interestingly, the tertiary structure of the anHIF-1α protein was found to be extremely similar to that of mice. Multiple comparison and phylogenetic tree results demonstrated that anHIF-1α was highly conserved. Under normoxic conditions, anHIF-1α mRNA transcripts could be detected in all tissues examined with the highest expression level in the heart. With gradually decreasing oxygen concentrations, anHIF-1α mRNA level was upregulated significantly in the gill, liver, kidney, spleen, intestine, brain, and muscle tissues (P < 0.05). Similarly, anHIF-1α was expressed in all examined bighead carp tissues, and the results suggested that the upregulation of anHIF-1α at the transcriptional level may be an important stress response adaptation to hypoxia in bighead carp. Finally, based on the tertiary structure comparative analyses between anHIF-1α with mouse HIF-1α, we think the physiological function, and protein structure of HIF-1α could be compared between fish and mammal in the future.

Chromosomal study of Khawia abbottinae (Cestoda: Caryophyllidea): karyotype and localization of telomeric and ribosomal sequences after fluorescence in situ hybridization (FISH).

An original cytogenetic study combining classical karyotype analysis and modern fluorescence in situ hybridization using telomeric (TTAGGG)n and ribosomal sequences (18S rDNA) was performed in Khawia abbottinae (Cestoda, Caryophyllidea), a parasite of Chinese false gudgeon (Abbottina rivularis) from China. Analyses based on conventional Giemsa staining, DAPI, YOYO-1 dye, and silver (Ag) staining were also carried out. The karyotype is composed of eight pairs of metacentric and telocentric chromosomes (2n = 16, n=5m + 3t). Constitutive heterochromatin was mainly positioned at pericentromeric regions, and telomeric sequences (TTAGGG)n were restricted to the end of all chromosomes. In mitotic preparations stained with Giemsa, both homologues of chromosome pair 4 showed a distinct secondary constriction. FISH with rDNA probe confirmed that this secondary constriction contains a nucleolar organizer region (NOR). The process of spermatocyte meiosis and the dynamics of nucleolus degradation in dividing cell were scrutinized. The present study and its results enhance the limited knowledge on basic karyotype characteristics and 18S rDNA clusters location in caryophyllidean tapeworms.

Morphological variation in Myxobolus drjagini (Akhmerov, 1954) from silver carp and description of Myxobolus paratypicus n. sp. (Cnidaria: Myxozoa).

There is uncertainty in the identification of Myxobolus drjagini, the causative agent of silver carp twist disease, in the literature. An investigation of fish parasites in Lake Taihu, China, revealed several Myxobolus drjagini-like myxosporeans infecting the subcutaneous tissue of the head skin, the olfactory and oculomotor nerves in the cranial cavity, and the intrafilamental epithelium of the gills of silver carp Hypophthalmichthys molitrix (Valenciennes, 1844). Myxospores from the head skin and the nerves were identified as conspecific to M. drjagini based on morphological and molecular data; although the spores from each of the two organs presented morphological variations. SSU rDNA sequence analysis revealed that the sequence of M. drjagini previously deposited in GenBank (AF085179) was invalid. Myxospores from the gills were identified as Myxobolus paratypicus n. sp. The spores were oval, asymmetric in frontal view, 13.8 (12.9-14.9) µm long, 9.9 (9.2-11.1) µm wide, and 7.0 µm thick. Two pyriform polar capsules were unequal in size (ratio above 4:1) with slightly converging anterior ends, and the posterior end of the large polar capsule extended beyond the middle of the spore. The large polar capsule was 7.5 (6.2-8.2) µm long and 5.0 (4.2-5.6) µm wide; the small polar capsule was 2.7 (2.1-3.6) µm long and 1.4 (1.1-1.9) µm wide. Polar filaments were coiled with 7-8 turns in the large polar capsule. The SSU rDNA sequence of M. paratypicus n. sp. was not identical to that of any myxozoan available in GenBank and showed highest similarity with M. drjagini (96%) and Myxobolus pavlovskii (95%) collected from bighead carp and silver carp, respectively.

Characterization of the complete mitochondrial genome of Parabreviscolexniepini Xi et al., 2018 (Cestoda, Caryophyllidea).

Parabreviscolexniepini is a recently described caryophyllidean monozoic tapeworm from schizothoracine fish on the Tibetan Plateau. In the present study, the complete mitochondrial genome of P.niepini is determined for the first time. The mitogenome is 15,034 bp in length with an A+T content of 59.6%, and consists of 12 protein-encoding genes, 22 tRNA genes, two rRNA genes, and two non-coding regions. The secondary structure of tRNAs exhibit the conventional cloverleaf structure, except for trnS1 (AGN) and trnR, which lack DHU arms. The anti-codon of trnS1 (AGN) in the mitogenome of P.niepini is TCT. The two major non-coding regions, 567 bp and 1428 bp in size, are located between trnL2 and cox2, trnG and cox3, respectively. The gene order of P.niepini shows a consistent pattern with other caryophyllideans. Phylogenetic analysis based on mitogenomic data indicates that P.niepini has a close evolutionary relationship with tapeworms Breviscolexorientalis and Atractolytocestushuronensis.

Tour around the globe: The case of invasive tapeworm Atractolytocestus huronensis (Cestoda: Caryophyllidea), a parasite of common carp.

The monozoic tapeworm Atractolytocestus huronensis Anthony, 1958 (Cestoda: Caryophyllidea), an intestinal parasite of the common carp, is characterized by its invasive character and potential to colonize new territories. It was initially described from North America and has also been found in several European countries. The most recent findings of A. huronensis originated from China and South Africa; however, no data on genetic relationships of these populations were available. The current study provides the first molecular characterisation of A. huronensis from South Africa and China using a partial sequence of mitochondrial cytochrome c oxidase subunit 1 (cox1) and a complete ribosomal ITS2 spacer. Ribosomal and mitochondrial data were applied for phylogenetic analyses in order to assess the genetic interrelationships among global A. huronensis populations. Divergent intragenomic copies of ribosomal ITS2 were detected in all analysed specimens; the structure and frequency of the ITS2 variants of tapeworms from China and South Africa corresponded with the data on ITS2 paralogues observed previously in A. huronensis from Slovakia, the United States and the United Kingdom. The phylogenetic analysis of cox1 indicated that A. huronensis exist in two slightly differentiated clusters; one cluster was supported by all phylogenetic approaches (NJ, ML, BI) and was represented by samples from China, the USA and the UK. A second cluster was represented by tapeworms from continental Europe (Slovakia, Hungary, Romania, Croatia) and South Africa. Haplotype network analysis revealed that the highest population diversity occurs in China. The results provide useful pilot information about the interrelationships of A. huronensis on four continents and indicate that China, or the eastern Palaearctic, served as the original source population for the global expansion of this invasive tapeworm. Data on the origin and distribution of the common carp, the only specific host of A. huronensis, are also discussed.

Effect of nitrite exposure on the antioxidant enzymes and glutathione system in the liver of bighead carp, Aristichthys nobilis.

Nitrite (NO2-) can cause oxidative stress in aquatic animal when it accumulates in the organism, resulting in different toxic effects on fish. In the present study, we investigated the effects of nitrite exposure on the antioxidant enzymes and glutathione system in the liver of Bighead carp (Aristichthys nobilis). Fish [Initial average weight: (180.05 ±â€¯0.092) g] were exposed to 48.634 mg/L nitrite for 96 h, and a subsequent 96 h for the recovery test. Fish livers were collected to assay antioxidant enzymes activity, hepatic structure and expression of genes after 0 h, 6 h, 12 h, 24 h, 48 h, 72 h, 96 h of exposure and12 h, 24 h, 48 h, 72 h, 96 h of recovery. The results showed that the activity of glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), and glutathione reductase (GR) increased significantly in the early stages of nitrite exposure. The study also showed that nitrite significantly up-regulated the mRNA levels of glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), and glutathione reductase (GR) after 6, 48, and 72 h of exposure respectively. Nitrite also increased the formation of malondialdehyde (MDA), oxidized glutathione (GSSG), and the activity of catalase (CAT). Nitrite was observed to reduce the activity of superoxide dismutase (SOD) and the level of glutathione (GSH). In the recovery test, GSH and the GSSG recovered but did not return to pre-stress levels. The results suggested that the glutathione system played important roles in nitrite-induced oxidative stress in fish. The bighead carp responds to oxidative stress by enhancing the activity of GSH-Px, GST, GR and up-regulating the expression level of GSH-Px, GST, GR, a whilst simultaneously maintaining the dynamic balance of GSH/GSSG. CAT was also indispensable. They could reduce the degree of lipid peroxidation, and ultimately protect the body from oxidative damage.

A new monozoic tapeworm, Parabreviscolex niepini n. g., n. sp. (Cestoda: Caryophyllidea), from schizothoracine fishes (Cyprinidae: Schizothoracinae) in Tibet, China.

A new monozoic cestode, Parabreviscolex niepini n. gen. and n. sp. (Cestoda: Caryophyllidea), is described from the type-host Schizopygopsis younghusbandi Regan, 1905 (Cyprinidae: Schizothoracinae) and Schizothorax waltoni Regan, 1905 (Cyprinidae: Schizothoracinae) in the Yarlung Tsangpo River, the upper tributary of the Brahmaputra River on the Tibetan Plateau. The new genus is placed in the Capingentidae because the vitellarium is situated partly in the medullary and cortical parenchyma, i.e., neither completely external nor internal to inner longitudinal muscles. Parabreviscolex n. gen. is characterized by possessing an afossate and cuneiform scolex; numerous vitelline follicles and testes present immediately after the scolex, and spread backward near the cirrus sac; the uterus does not loop anterior to the cirrus sac; genital pores separate, opening to the common genital atrium; the pre-ovarian vitelline follicles lateral and median, post-ovarian vitelline follicles present; ovary H-shaped, compact, and ovarian arms long, anteriorly reaching the cirrus sac. Homology search by the basic local alignment search tool (BLAST) showed that the partial 18S rDNA and complete mtDNA cox-1 sequences obtained in this report were not consistent with any sequences available in GenBank, and molecular phylogenetic analyses revealed Parabreviscolex formed a separated long branch within the caryophyllideans from cyprinids.

Myxobolus linzhiensis n. sp. (Myxozoa: Myxobolidae) from the gill filament of Schizothorax oconnori Lloyd (Cyprinidae: Schizothoracinae) in Tibet, China: morphological and molecular characterization.

In this report, a new myxosporean species, Myxobolus linzhiensis n. sp. (Cnidaria: Myxozoa: Bivalvulida), was described from the endemic cyprinid Schizothorax oconnori inhabiting the Yarlung Tsangpo River in the Tibetan plateau, China. The whitish ellipsoidal plasmodia of M. linzhiensis n. sp. grew under the epithelium of gill filaments, 1.7-mm long and 0.4-mm wide, and were found in the 3 of 8 examined S. oconnori (37.5%). Mature spores appeared suborbicular in frontal view, fusiform shaped in lateral view with tapering anterior, measuring 11.4 ± 0.7 (10.1-13.0) µm in length, 10.6 ± 0.4 (10.0-11.3) µm in width, and 6.6 ± 0.3 (6.2-7.0) µm in thickness. Two equal polar capsules were pyriform with an apophysis at its top end, measuring 5.7 ± 0.5 (4.8-6.7) µm in length, 3.8 ± 0.2 (3.2-4.2) µm in width, and polar filaments coiled 4 turns. Spore valves were symmetrical, and sutural ridge was straight and thin, running near the middle of the valves. Although Myxobolus linzhiensis n. sp. showed similar morphological characteristics with Myxobolus chushi Dar Kaur & Chishti 2017 from Schizothorax niger, Myxobolus kienweiensis Ma, 1976 from Schizothorax davidi, the new myxosporean was distinguished with them in spore shape, polar filament, and host-tissue tropism. Homology search by BLAST in GenBank indicated that the obtained 18S rDNA sequence of M. linzhiensis n. sp. (KY965935) did not match any available myxozoan sequence, most similar to Henneguya zikaweiensis, and showing less than 93% sequence similarity. Phylogenetical analyses demonstrated that M. linzhiensis n. sp. was firmly clustered in the clade consisting of myxosporeans Thelohanellus, Myxobolus, Henneguya from Asian cyprinids. This study is the first report on myxosporean parasitizing in endemic fish in the Tibetan Plateau, China.

Description of a new Neoactinomyxum type actinosporean from the oligochaete Branchiura sowerbyi Beddard.

Actinosporean infection of oligochaetes living in the mud of a commercial gibel carp pond with myxosporean disease was studied. Six actinospore types were detected exclusively from the oligochaete Branchiura sowerbyi Beddard with very high prevalence (18%). Five out of the six types were identified as the same actinosporeans described in previous reports, the sixth actinosporean was identified as a new Neoactinomyxum type and described here based on morphological and molecular characterisation. Spore body of the actinospore was globular, much smaller than caudal processes. Three caudal processes were disc-like in apical view, hemispherical in side view, closer together and encircling the spore body. The number of sporoplasm cells was detected as eight in one specimen. The new actinosporean markedly differed from other Neoactinomyxum types in literature having much bigger caudal processes. DNA sequence analyses further confirmed the morphological identification, and revealed the actinosporean described here (KU641392) possessed less than 94% sequence similarity with myxozoans available in the GenBank database.

The occurrence of the common European fish cestode Caryophyllaeus laticeps (Pallas, 1781) in the River Irtysh, China: a morphological characterization and molecular data.

The cestode Caryophyllaeus laticeps (Pallas, 1781) is a highly polymorphic species with several morphotypes, and belongs to the most widespread species of monozoic tapeworm (Cestoda: Caryophyllidea) found in the Palaearctic zoogeographic region with a wide range of cyprinid fish hosts. This paper provides the first reliable evidence about the occurrence of C. laticeps in northern China that is based on detailed morphological characteristics and mitochondrial cox1 sequences. C. laticeps was found in freshwater bream Abramis brama (L.), in the River Irtysh, with the prevalence of 40% and mean intensity of infection 8. The tapeworms were rather similar in their morphological characters to the morphotype 1 of C. laticeps which is typified by the possession of a slender body with a flabellate wide scolex, narrow neck, robust, long and clearly visible vas deferens, large, and almost spherical cirrus-sac containing well-developed internal seminal vesicle and the first vitelline follicles and first testes beginning far from anterior margin of the body. Subsequent molecular analysis confirmed the morphological identification of the species. Our results show that the studied specimens were firmly clustered into the C. laticeps clade and formed a monophyletic group with C. laticeps morphotype 1 and 2 from Europe and Russia. In China, C. laticeps is a non-indigenous fish parasite, which has the most probably been introduced into the Irtysh River basin with its typical host inhabiting Russia.

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.

Development of highly sensitive electrochemical genosensor based on multiwalled carbon nanotubes-chitosan-bismuth and lead sulfide nanoparticles for the detection of pathogenic Aeromonas.

In this paper, we reported the construction of new high sensitive electrochemical genosensor based on multiwalled carbon nanotubes-chitosan-bismuth complex (MWCNT-Chi-Bi) and lead sulfide nanoparticles for the detection of pathogenic Aeromonas. Lead sulfide nanoparticles capped with 5'-(NH2) oligonucleotides thought amide bond was used as signalizing probe DNA (sz-DNA) and thiol-modified oligonucleotides sequence was used as fixing probe DNA (fDNA). The two probes hybridize with target Aeromonas DNA (tDNA) sequence (fDNA-tDNA-szDNA). The signal of hybridization is detected by differential pulse voltammetry (DPV) after electrodeposition of released lead nanoparticles (PbS) from sz-DNA on the surface of glass carbon electrode decorated with MWCNT-Chi-Bi, which improves the deposition and traducing electrical signal. The optimization of incubation time, hybridization temperature, deposition potential, deposition time and the specificity of the probes were investigated. Our results showed the highest sensibility to detect the target gene when compared with related biosensors and polymerase chain reaction (PCR). The detection limit for this biosensor was 1.0×10(-14) M. We could detect lower than 10(2) CFU mL(-1) of Aeromonas in spiked tap water. This method is rapid and sensitive for the detection of pathogenic bacteria and would become a potential application in biomedical diagnosis, food safety and environmental monitoring.

Molecular evidence of cryptic diversity in Paracaryophyllaeus (Cestoda: Caryophyllidea), parasites of loaches (Cobitidae) in Eurasia, including description of P. vladkae n. sp.

Molecular phylogenetic analysis of an extensive collection of monozoic tapeworms of the genus Paracaryophyllaeus Kulakovskaya, 1961 (Cestoda: Caryophyllidea), parasites of loaches (Cypriniformes: Cobitidae) in Eurasia, has revealed cryptic species diversity within this long-time monotypic genus, especially in the Paracaryophyllaeus gotoi (Motomura, 1927) species complex [syn. Paracaryophyllaeus dubininorum (Kulakovskaya, 1961); type species]. Three independent, well-supported clades were discovered on the basis of molecular data: (i) specimens from Misgurnus anguillicaudatus and Cobitis lutheri from China, Russian Far East and Japan - called herein P. cf. gotoi 1, which may be conspecific with P. gotoi (Motomura, 1927), although in the absence of sequence data for P. gotoi from its type locality (basin of the River Kumkan in Korea), no certain inferences about their identity can currently be made; (ii) specimens from M. anguillicaudatus from China and Japan - P. cf. gotoi 2, which are morphologically indistinguishable from those of P. cf. gotoi 1; and (iii) morphologically distinct tapeworms from the endemic loach Cobitis bilseli from southwestern Turkey (Beysehir Lake), which are described herein as a new species. Paracaryophyllaeus vladkae Scholz, Oros and Aydogdu n. sp. differs from the remaining species of the genus in the following characteristics: the testes begin anterior to the first vitelline follicles (versus posterior), the body is short and robust (versus more elongate and slender), and the scolex is wide, rounded or apically tapered (versus claviform to truncate). Species composition of the genus, host specificity of species and geographical distribution are briefly discussed.

Khawia abbottinae sp. n. (Cestoda: Caryophyllidea) from the Chinese false gudgeon Abbottina rivularis (Cyprinidae: Gobioninae) in China: morphological and molecular data.

Khawia abbottinae sp. n. is described from the Chinese false gudgeon, Abbottina rivularis (Basilewsky) (Cyprinidae: Gobioninae), from the Yangtze River basin in China. The new species can be distinguished from the congeneric species mainly by the arrangements of the testes, which form two longitudinal bands (other congeneric species have the testes irregularly scattered throughout the testicular region) and their number (at maximum 85 testes versus at least 160 in the other Khawia spp.), and the morphology of the scolex, which varies from cuneiform to widely bulbate scolex, being separated from the remaining body by a short neck and possessing a smooth, blunt or rounded anterior margin. Other typical features of K. abbottinae are its small size (total length less than 1.5 cm) and body shape, with the maximum width at its first third. The distinct status of the new species was confirmed by molecular data (ssrDNA and ITS1 sequences). Phylogenetic analyses revealed a close relationship of the new species with K. rossittensis (Szidat, 1937) and K. parva (Zmeev, 1936), parasites of crucian carp and goldfish (Carassius spp.), but both species markedly differ from K. abbottinae in their morphology. Until now, five valid species of Khawia (K. abbottinae, K. japonensis, K. rossittensis, K. saurogobii and K. sinensis) have been reported from China.

Three actinosporean types (Myxozoa) from the oligochaete Branchiura sowerbyi in China.

Three actinosporean types--raabeia, aurantiactinomyxon, and guyenotia--from the oligochaete Branchiura sowerbyi Beddard collected from a crucian carp (Carassius auratus gibelio) pond are described in this report. Compared with the actinospores described previously, the raabeia type presents similar spore shape with the actinospore of Myxobolus cultus, except a much longer caudal process, 250.8 µm (217.5-276.3). The aurantiactinomyxon and guyenotia type significantly differ from other actinospores in its same collective group with different caudal process shape (taper and leaf-like) and dimension, 170.8 µm (167.5-176.3) and 18.5 µm (16.5-20.6), respectively. The partial 18S rDNA sequences of raabeia and aurantiactinomyxon types, and myxospores M. cultus, Myxobolus wulii, Myxobolus pyramidis, and Thelohanellus wuhanensis detected from pond-reared crucian carp were determined. Based on DNA sequences analysis, the raabeia type showed high genetic similarity (99.5-100%) with myxospore M. cultus Yokoyama, Ogawa & Wakabayashi, 1995 on the gills of crucian carp sampled from the same pond. The aurantiactinomyxon showed no more than 85.2% sequence similarity with myxospores determined in this report and other myxozoans available from GenBank.

Distribution and virulence gene comparison of Aeromonas strains isolated from diseased fish and water environment.

A total of 71 Aeromonas strains were isolated in the south of Jiangsu Province China in order to analyze the difference ofAeromonas spp. distribution between diseased fish and water environment. The sequence of 16S rDNA and gyrB demonstrated that the 71 Aeromonas isolates could be divided into 4 species, including A. veronii (55), A. hydrophila (11), A. salmonicida (3) and A. media (2). A. veronii was the most common species isolated from fish and water environment. All Aeromonas isolates were screened for three putative virulence genes, aer, hly and alt. hly was the most common gene among three virulence genes.

A chromosome study and localization of 18S rDNA in Khawia saurogobii (Cestoda: Caryophyllidea).

This paper reports results of the first cytogenetic study carried out on a recently described monozoic tapeworm, Khawia saurogobii Xi et al., 2009, from the Chinese lizard gudgeon (Saurogobio dabryi). The karyotype of this species is composed of eight pairs of metacentric and telocentric chromosomes (2n = 16; n = 3m + 5t), metacentric chromosomes representing the first, sixth, and eight pairs. All chromosomes except the largest pair displayed 4',6-diamidino-2-phenylidole (DAPI) positive heterochromatin in centromeric regions. In mitotic preparations stained with Giemsa, one of the homologues of a smaller metacentric chromosome pair (No. 7) showed a distinct secondary constriction, whereas the other did not. Fluorescent in situ hybridization (FISH) with 18S ribosomal DNA (rDNA) probe revealed that the chromosomes No. 7 carry each a cluster of ribosomal genes associated with the centromeric heterochromatin and confirmed that this chromosome pair contains a nucleolar organizer region (NOR). The rDNA-FISH also confirmed heteromorphism in the size of NOR (i.e., secondary constriction) observed after Giemsa staining. The present cytogenetic analysis revealed species-specific characters of K. saurogobii and showed that FISH may represent a new valuable cytogenetic tool suitable for comparative taxonomic or phylogenetic studies within the order Caryophyllidea in the future.