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The neuropeptide B/W signaling system is composed of neuropeptide B (NPB), neuropeptide W (NPW), and two cognate receptors, NPBWR1 and NPBWR2, which are involved in diverse physiological processes, including the central regulation of neuroendocrine axes in vertebrates. The components of this signaling system are not well conserved during vertebrate evolution, implicating its functional diversity. The present study characterized the ricefield eel neuropeptide B/W system, generated a specific antiserum against the neuropeptide B/W receptor, and examined the potential roles of the system in the regulation of adenohypophysial functions. The ricefield eel genome contains npba, npbb, and npbwr2b but lacks the npw, npbwr1, and npbwr2a genes. The loss of npw and npbwr1 probably occurred at the base of ray-finned fish radiation and that of npbwr2a species specifically in ray-finned fish. Npba and npbb genes are produced through whole-genome duplication (WGD) in ray-finned fish. The ricefield eel npba was expressed in the brain and some peripheral tissues, while npbb was predominantly expressed in the brain. The ricefield eel npbwr2b was also expressed in the brain and in some peripheral tissues, such as the pituitary, gonad, heart, and eye. Immunoreactive Npbwr2b was shown to be localized to Lh and Fsh cells but not to Gh or Prl cells in the pituitary of ricefield eels. Npba upregulated the expression of fshb and cga but not lhb mRNA in pituitary fragments of ricefield eels cultured in vitro. The results of the present study suggest that the NPB system of ricefield eels may be involved in the neuroendocrine regulation of reproduction. [ABSTRACT FROM AUTHOR]

Primordial germ cells (PGCs) play an important role in sexual fate determination and gonadal development in gonochoristic fish, such as zebrafish and medaka. However, little is known about the function of PGCs in hermaphroditic fish. Rice field eel (Monopterus albus), a protogynous hermaphroditic fish, is an economically valuable aquaculture species. We eliminated PGCs in rice field eels during embryogenesis via morpholino-mediated knockdown dead end (dnd). The PGCs-depleted gonads developed into testis-like structures with Sertoli cells and Leydig cells. The gene expression pattern of 15-month-old PGCs-depleted gonads showed that male-biased genes, dmrt1, sox9a, gsdf, and amh, were significantly higher than that of the WT, whereas female-biased genes, foxl2 and cyp19a1a, were significantly decreased. These results indicate that PGCs are essential for ovarian differentiation in rice field eel, and PGCs-depleted gonads develop into sterile males without undergoing the female and intersex stages. Our study is the first to identify the role of PGCs in sex differentiation in rice field eel, a protogynous hermaphrodite teleost. And it is of great significance in rice field eel for discovering the underlying mechanism of sex differentiation and establishing sex control technology. [ABSTRACT FROM AUTHOR]

Discoveries of new Hemiclepsis species (Hirudinea: Glossiphoniidae) in East and Southeast Asia were expected. A peculiar freshwater leech was found on the body surface of Asian Swamp Eel Monopterus albus (Synbranchiformes: Synbranchidae) in Jingzhou, Hubei Province, China. Here, we describe this leech species as Hemiclepsis yangtzenensissp. nov. It could be distinguished from other congeners based on a combination of morphological features such as very small size, translucent body with dense green reticulate markings throughout dorsum, very large posterior sucker with a characteristic reticulate pattern, and two pairs of well-developed drop-like eyespots. It also represents a phylogenetic lineage that is distant from other members of the genus based on the COI and 18S rRNA sequences. This leech species seems to be a specialized fish parasite that could negatively affect farming and wild stocks of Asian Swamp Eel in China. [ABSTRACT FROM AUTHOR]

Oxidative stress contributes a lot to initiation and progression of pathological conditions. Heme oxygenase 1 (HO1), a cytoprotective enzyme, is usually upregulated to alleviate oxidative stress in vivo. The function of teleost HO1 in the response to oxidative stress induced by heavy metal exposure and in pathogenic bacterial infection remains uncertain. In the present study, both complementary DNA and genomic sequence of a HO1-like gene cloned from the liver of swamp eel (Monopterus albus) are reported. Sequence analysis showed that the putative amino acid sequence contained a conserved heme oxygenase signature and displayed higher similarity to HO1 genes of other teleosts. Expression profile of swamp eel HO1 was investigated in healthy tissues and in tissues following stimulation with pathogenic bacteria (Aeromonas hydrophila) or cadmium chloride (CdCl2) exposure. Results demonstrated that HO1 messenger RNA (mRNA) was highly expressed in the liver and relatively less in other tissues. Bacterial infection with A. hydrophila significantly changed HO1 mRNA expression in the liver, spleen, and kidney, and the mRNA expression of HO1 and Nrf2 in the liver was elevated after the fish were exposed to CdCl2. Subsequently, the swamp eel HO1 was subcloned into a pET28a expression vector and transformed into Escherichia coli BL21 (DE3). Recombinant HO1 (rHO1) was successfully induced by 0.1 mmol/l IPTG and purified by Ni-NTA His Bind Resin purification system. To determine whether the rHO1 could confer stress tolerance in vitro, the viability of control and HO1-expressing E. coli under CdCl2 stress was compared by spot assay. The rHO1 protein significantly increased survival rates of the bacterial hosts. To evaluate whether intraperitoneal injection with rHO1 protected the liver of swamp eel against A. hydrophila-induced oxidative stress, mRNA expression of HO1, Nrf2, hepcidin, and IL-1β as well as the oxidative stress-related parameters (ROS and total antioxidant capacity (T-AOC)) in the liver were examined. The results showed that exogenous rHO1 could significantly upgrade the mRNA expression of HO1 and hepcidin, coupled with increased ROS and T-AOC levels. However, Nrf2 and IL-1β expression levels were significantly downregulated and upregulated, respectively. These results suggested that HO1 should not only play a protective role in oxidative stress response and its adverse effects deserved further investigation. [ABSTRACT FROM AUTHOR]

Smad2, a receptor-activated Smad, plays a critical role in regulating gametogenesis. In this study, a smad2 homologue was identified and sequenced from ricefield eel ovary cDNA, and its mRNA and protein expression levels were analysed during oocyte development. The cDNA sequence of ricefield eel smad2 consisted of 1863 bp encoding a 467-amino acid protein that had high sequence homology with Smad proteins in other teleosts, especially in Poeciliopsis prolifica. The results of real-time quantitative PCR (RT-qPCR) analysis revealed that smad2 is expressed in the ovary during gonad development, increased continuously until the early vitellogenic stage in the ovaries, and then decreased with ovary maturation. Smad2 protein immunoreactivity was localized in the cytoplasm of follicular cells, oogonia, and primary growth stage oocytes. In vitro experiments revealed that follicle-stimulating hormone (FSH) and human chorionic gonadotropin (hCG) promoted smad2 expression in ovary tissue in a time- and dose-dependent manner, respectively. In summary, Smad2 plays a potentially vital role in ricefield eel ovary development. [ABSTRACT FROM AUTHOR]

In 2017, a clinical disease outbreak resulted in substantial mortality of adults and larvae of cultured Chinese rice-field eels (Monopterus albus) on a farm in Hubei, Central China. A rhabdovirus was isolated from moribund specimens, and typical clinical symptoms associated with an outbreak included an enlarged and swollen head. This differed from previous observations. Histological changes included necrosis and cavities of various sizes within the brain and kidney. Homogenized tissues of diseased Chinese rice-field eels were screened for viral isolation using six different fish cell lines. A rhabdovirus was isolated following observation of cytopathic effect (CPE) in a gibel carp brain (GiCB) cell line and confirmed by RT-PCR. Electron microscopy showed large numbers of rhabdovirus-shaped particles in the cytoplasm of the brain cells of the diseased Chinese rice-field eels and in the infected GiCB cell line. This virus has been named "Chinese rice-field eel rhabdovirus" (CrERV), and the complete nucleotide sequence of CrERV was cloned. This rhabdovirus is composed of 11,545 nucleotides with the following genomic organization: 3′-N-P-M-G-L-5′. The genes are separated by conserved gene junctions, and phylogenetic analysis of the L sequence revealed that CrERV forms a separate branch with Siniperca chuatsi rhabdovirus (SCRV) and hybrid snakehead rhabdovirus C1207 (HSHRV-C1207). This is the first report of the complete sequence of CrERV from the Chinese rice-field eel in China. [ABSTRACT FROM AUTHOR]

Interleukin-1β (IL-1β) is the prototypic pro-inflammatory cytokine, whose functions are mediated through interaction with its receptors (IL-1R1 and IL-1R2). Herein, we cloned the full-length cDNA and genomic DNA of IL-1β and IL-1R2 in the Asian swamp eel ( Monopterus albus). The eel IL-1β cDNA encodes a putative polypeptide of 246 amino acids. The protein sequence includes a typical IL-1 family signature, but lacked an interleukin-converting enzyme cleavage site. The genomic DNA of eel IL-1β was 2520 bp and comprised five exons and four introns. The eel IL-1R2 cDNA encoded a putative propeptide of 423 amino acid residues, comprising a signal peptide, a transmembrane region and two Ig-like domains in the extracellular region. Similar to other vertebrates, the genomic DNA of the eel IL-1R2 has nine exons and eight introns. Real-time PCR analysis indicated that IL-1β and IL-1R2 were constitutively expressed in all tissues, especially in the liver and immune-related organs. After infection with Aeromonas hydrophila, the transcript levels of IL-1β and IL-1R2 were induced in the head kidney and spleen, reaching their highest levels at 6 h post injection. In vitro, IL-1β and IL-1R2 mRNA levels were also upregulated rapidly at 1h post infection with A. hydrophila. Furthermore, acanthocephalan Pallisentis (Neosentis) celatus could induce the expression of both genes in the head kidney and intestine. In infected intestines, the transcript levels of IL-1β and IL-1R2 were increased by 21.4-fold and 20.8-fold, respectively, relative to the control. The present study indicated that IL-1β and IL-1R2 play an important role in inflammation and host defense, especially in the antiacanthocephalan response. [ABSTRACT FROM AUTHOR]

We isolated the complete Foxl2 (Foxl2a) cDNA from the Monopterus albus ovary. An alignment of known Foxl2 amino-acid sequences confirmed the conservation of the Foxl2 open reading frame, especially the forkhead domain and C-terminal region. The expression of Foxl2 was detected in the brain, eyes, and gonads. A high level of Foxl2 expression in the ovary before sex reversal, but its transcripts decreased sharply when the gonad developed into the ovotestis and testis. The correlation between the Foxl2 expression and the process of sex development revealed the important function of Foxl2 during the sex reversal of M. albus. Immunohistochemical analysis showed that Foxl2 was expressed abundantly in granulosa cells and in the interstitial cells of the ovotestis and testis. These results suggest that Foxl2 plays a pivotal role in the development and maintenance of ovarian function. Foxl2 may be also involved in the early development of testis and the development of ocular structures of M. albus. [ABSTRACT FROM AUTHOR]

A 10-week feeding experiment was conducted to evaluate the effects of dietary protein to lipid ratios (P/L) on growth, intestinal digestive enzyme activities and body composition in juvenile rice field eel ( Monopterus albus) (initial mean body weight of 65.76 ± 1.07 g, mean ± SEM). Nine test diets were formulated in a 3 × 3 factorial design to contain three protein levels (350, 400 and 450 g kg) for each of three lipid levels (40, 80 and 120 g kg), respectively. Each diet was randomly assigned to triplicate groups of 60 fish per net cage (1.5 × 2.0 × 1.5 m). Results showed the survival was above 96 % and was not affected by dietary treatments. Fish fed the diet with 450 g kg protein and 40 g kg lipid showed the best weight gain (WG) (103.95 %) and feed conversion ratio (1.60) ( P < 0.05). WG, protein efficiency ratio and energy retention increased with the increasing in lipid at 350 g kg protein level ( P < 0.05). However, WG showed a little decline with increasing dietary lipid when fish fed the diets with 400 and 450 g kg protein level, but no significant difference was observed ( P < 0.05). Hepatosomatic index, visceralsomatic index and intestinal lipase activity increased with the increasing of dietary lipid level irrespective protein level. Intestinal trypsin activity increased with the increasing of dietary lipid level when fish fed the diets with 350 g kg protein, but showed converse trend when fish fed the diets with 400 and 450 g kg protein. Serum triglyceride, body lipid and energy were positively correlated with the dietary lipid. Results of the present study showed that the dietary protein/lipid ratio of 450/40 g kg is considered optimum for rice field eel under culture conditions, and the increase in dietary lipid level has no efficient protein-sparing effect when fish fed the diets with 400 and 450 g kg protein level. [ABSTRACT FROM AUTHOR]

Metal content was determined in two fish species ( Misgurnus anguillicaudatus and Monopterus albus) inhabiting rice paddies along the Middle Reach of the Yangtze River. Both are species important food fish in East Asia. The levels of eight metals (Fe, Mn, Zn, Cu, Cr, As, Cd, Pb) in skin, muscle and intestinal tissues were measured and correlated with fish size. Metal distribution among the tissues was examined and the potential risk of fish consumption was evaluated. Correlations between metal concentration and fish size were not significant. Concentrations of essential metals varied significantly among tissues, but the differences were not significant for non-essential metals. Correlations among metals in skin tissue were stronger than in intestinal and muscle tissues. Metal concentrations in the edible tissues of both fish met permissible levels for consumption, except Pb in loach skin and Cr loach in muscle. The potential risk caused by Pb from skin and Cr from muscle of loach consumption should be considered in future risk assessmeents. [ABSTRACT FROM AUTHOR]

Livers and muscles of swamp eels ( Monopterus albus) were analyzed for bioaccumulation of heavy metals during the plowing stage of a paddy cycle. Results showed heavy metals were bioaccumulated more highly in liver than muscle. Zinc (Zn) was the highest bioaccumulated metal in liver (98.5 ± 8.95 μg/g) and in muscle (48.8 ± 7.17 μg/g). The lowest bioaccumulated metals were cadmium (Cd) in liver (3.44 ± 2.42 μg/g) and copper (Cu) in muscle (0.65 ± 0.20 μg/g). In sediments, Zn was present at the highest mean concentration (52.7 ± 2.85 μg/g), while Cd had the lowest mean concentration (1.04 ± 0.24 μg/g). The biota-sediment accumulation factor (BSAF) for Cu, Zn, Cd and nickel (Ni) in liver tissue was greater than the corresponding BSAF for muscle tissue. For the three plowing stages, metal concentrations were significantly correlated between liver and muscle tissues in all cases, and between sediment and either liver or muscle in most cases. Mean measured metal concentrations in muscle tissue were below the maximum permissible limits established by Malaysian and U.S. governmental agencies, and were therefore regarded as safe for human consumption. [ABSTRACT FROM AUTHOR]

Myoglobin from Asian swamp eel Monopterus albus was purified from fish muscle using salt fractionation followed by column chromatography and molecular filtration. The purified Mb of 0.68 mg/g wet weight of muscle was determined for its molecular mass by MALDI-TOF-MS to be 15,525.18 Da. Using isoelectric focusing technique, the purified Mb showed two derivatives with pI of 6.40 and 7.12. Six peptide fragments of this protein identified by LC-MS/MS were homologous to Mbs of sea raven Hemitripterus americanus, yellowfin tuna Thunnus albacores, blue marlin Makaira nigicans, common carp Cyprinus carpio, and goldfish Carassius auratus. According to the Mb denaturation, the swamp eel Mb had thermal stability higher than walking catfish Clarias batrachus Mb and striped catfish Pangasius hypophthalmus Mb, between 30 and 60 C. For the thermal stability of Mb, the swamp eel Mb showed a biphasic behavior due to the O dissociation and the heme orientation disorder, with the lowest increase in both Kd and Kd. The thermal sensitivity of swamp eel Mb was lower than those of the other Mbs for both of fast and slow reaction stages. These results suggest that the swamp eel Mb globin structure is thermally stable, which is consistent with heat-tolerant behavior of the swamp eel particularly in drought habitat. [ABSTRACT FROM AUTHOR]

The ricefield eel ( Monopterus albus) is an economic fish species in China. To improve its artificial reproduction, we studied the compositions and functions of the hatching froth secreted by ricefield eels. The froth showed a viscosity of 2.72 mPaS, composed of glycoproteins with a sugar content of 0.156 mg/ml and a protein content of 0.250 mg/ml at a ratio of 1.60. The froth proteins were identified by SDS-PAGE as four proteins with the molecular weights of 42.5, 58.6, 65.3, and 98.2 kDa respectively. Their total amino acid content was 201.88 mg/100 ml, consisting of 16 kinds of amino acids. Filed observation revealed three beneficial effects of the froth on the hatching of the eel eggs. First, the froth increased the hatching rates of fertilized eggs at a range of water temperatures. Second, the froth decreased the rates of infection of the fertilized eggs by the water mold. Finally, the froth accelerated velum breakage in the fertilized eggs. Of the different hatching methods, froth incubation achieved the highest hatching rate and larvae survival rate. These results demonstrate the indispensable function of the froth of ricefield eels, and offer practical reasons for protecting the froth during the breeding season. [ABSTRACT FROM AUTHOR]

Recently, many new rice–fish co-culture models have been developed to increase economic and ecological benefits. In this study, we added eels (Monopterus albus) to a rice–crayfish system and conducted a 3-year field investigation to compare the yields and availability of fertilizer N among groups with a low density of eels, high density of eels and no eels. We performed a mesocosm experiment and used an isotope tracer technique to detect the fate of fertilizer N. The results showed that the rice yields significantly improved after the introduction of the eels. However, the introduction of a high density of eels significantly limited the crayfish yield, increased water N and N2O emissions and decreased soil N content. The mesocosm experiment suggested that the use efficiency of fertilizer N was significantly increased after the introduction of the eels. The fertilizer N used by rice was significantly higher in rice–crayfish–eel system than in rice–crayfish system. This study indicated that the introduction of eels may be a good practice for improving yields and availability of fertilizer N in a rice–crayfish system. [ABSTRACT FROM AUTHOR]

The full-length cDNAs of amh and dax1 in the hermaphrodite, rice-field eel (Monopterus albus), were cloned and characterized in this study. Multiple sequence alignment revealed Dax1 was well conserved among vertebrates, whereas Amh had a low degree of similarity between different vertebrates. Their expression profiles in gonads during the course of sex inversion and tissues were investigated. The tissue distribution indicated amh was expressed mostly in gonads and was scarcely detectable in other tissues, whereas the expression of dax1 was widespread among the different tissues, especially liver and gonads. amh was scarcely detectable in ovaries whereas it was abundantly expressed in both ovotestis and testis. By contrast, dax1 was highly expressed in ovaries, especially in ♀IV (ovaries in IV stage), but it was decreased significantly in ♀/♂I (ovotestis in I stage). Its expression was increased again in ♀/♂III (ovotestis in III stage), and then decreased to a low level in testis. These significant different expression patterns of amh and dax1 suggest the increase of amh expression and the decline of dax1 expression are important for the activation of testis development, and the high level of amh and a low level of dax1 expression are necessary for maintenance of testis function. [ABSTRACT FROM AUTHOR]