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1.
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2.
Enteromyxosis caused by Enteromyxum scophthalmi is one of the parasitizations with a higher economic impact on turbot, Scophthalmus maximus (L.), aquaculture. This myxosporean produces severe catarrhal enteritis with abundant inflammatory infiltrates in the lamina propria‐submucosa (LP), epithelial detachment and leucocyte depletion of the lymphohaematopoietic organs. Some advances made on the pathogenesis pointed to a role of apoptosis in the enteromyxosis. Therefore, the main aim of this work was to employ the TUNEL assay and the anti‐(active caspase‐3) immunohistochemical assay to detect apoptotic cells in both healthy and E. scophthalmi‐infected turbot in order to establish the presence and distribution of apoptotic cells during development of the disease. More apoptotic cells located within the gastrointestinal epithelium were observed in the initial stages of the infection in E. scophthalmi‐infected turbot compared with non‐infected turbot. As the infection progressed, a higher degree of apoptosis occurred in the epithelium of folds heavily parasitized. In the severely infected turbot, apoptosis was also found among the leucocytes of the intestinal inflammatory infiltrates. Moreover, the number of active caspase‐3‐positive cells in the lymphohaematopoietic organs tended to increase with disease severity. In view of the results, increased apoptosis in the epithelium may favour the scaling that occurs during enteromyxosis and cell death of leucocytes in the intestinal LP, contributing to leucocyte depletion in severe cases.  相似文献   

3.
Identification of fish larval behavioural traits permitting capture of specific live prey sizes is an important part of optimizing production of marine larvae. We investigated the capture success of turbot larvae (Scophthalmus maximus) at two development stages, 8 and 10 days post‐hatch (DPH), when offered small nauplii (129–202 μm), large nauplii (222–278 μm) and copepodites (342–542 μm), of the calanoid copepod Acartia tonsa. At 8 DPH, turbot larvae had the highest capture success (67%) when offered small nauplii, with a lower capture success of large nauplii (27%) but totally lacked the capabilities to capture copepodites. At DPH 10, the larvae increased the capture success of large nauplii (47%) and achieved a few successful attacks on copepodites. Energetically, large nauplii were the most beneficial at both larval development stages. The swimming kinematics of the period prior to a strike by the larva on the copepod was examined, and the approach pattern of the larva was identified as a controlling mechanism for their strike distance, with the initial approach speed of larva at DPH 10 being significantly less than at DPH 8. In all successful attacks, the strike distance was less than 1.17 mm and was significantly lower than unsuccessful attacks. Since the approach pattern of the larva is linked to its capture success, it could be used as the basis for a feeding scheme based on the swimming performance of individual batches of turbot larvae.  相似文献   

4.
An outbreak of disease characterized by skin ulcers, fin rot and mortality was observed a few days after the transfer of Atlantic salmon (Salmo salar) from a freshwater smolt production facility to a land‐based seawater post‐smolt site. Dead and moribund fish had severe skin and muscle ulcers, often 2–6 cm wide, particularly caudal to the pectoral fins. Microscopic examination of smears from ulcers and head kidney identified long, slender Gram‐negative rods. Histopathological analysis revealed abundance of long, slender Tenacibaculum‐like bacteria in ulcers and affected fins. Genetic characterization using multilocus sequence analysis (MLSA) of seven housekeeping genes, including atpA, dnaK, glyA, gyrB, infB, rlmN and tgt, revealed that the isolates obtained during the outbreak were all clustered with the Tenacibaculum dicentrarchi‐type strain (USC39/09T) from Spain. Two bath challenge experiments with Atlantic salmon and an isolate of T. dicentrarchi from the outbreak were performed. No disease or mortality was observed in the first trial. In the second trial with a higher challenge dose of T. dicentrarchi and longer challenge time, we got 100% mortality within 48 hr. This is the first reported outbreak of disease caused by T. dicentrarchi in Norwegian farmed Atlantic salmon.  相似文献   

5.
The present study was conducted to evaluate the effects of dietary Eucommia ulmoides (EU) on growth, feed utilization, antioxidant activity and immune responses of turbot fed a basal diet (CON) or EU‐supplemented diets with 5.0 g/kg (EU1), 10.0 g/kg (EU2) and 20.0 g/kg (EU3) EU leaf powder. After 70‐day trial, EU supplementation did not affect nutrient utilization, but reduced feed intake (FI) and specific growth rate (SGR) of fish at doses above 5.0 g/kg. Superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (T‐AOC) activities in the EU‐supplemented groups were significantly higher than those in the control group at a dose of 20.0 g/kg. Significantly reduced malondialdehyde (MDA) contents were observed in EU‐supplemented groups at doses over 5.0 g/kg. Furthermore, fish fed 20.0 g/kg EU showed the highest lysozyme (LZM) activity among groups. The EU‐supplemented diets with doses above 5.0 g/kg significantly enhanced the mRNA expressions of cytokines. The expression level of major histocompatibility complex II alpha (MHC IIα) was significantly upregulated compared to that of the control fish when the supplemental level was at 20.0 g/kg. Taken together, the present study indicated that the EU could remarkably enhance the antioxidant activity, non‐specific immunity and maintain an active immune response in turbot.  相似文献   

6.
Enteromyxum scophthalmi is a myxozoan parasite that causes severe parasitic diseases in cultured turbot affecting mainly the intestine of the host. It is characterized by producing acute enteritis, starvation and eventually death. Current diagnosis of E. scopthalmi use traditional techniques, based on the identification of the morphology of the parasite. These techniques take extended time to be carried out and do not favour the adoption of control measure at turbot farms and require the sacrifice of fish. This study develops a fast real‐time PCR molecular tool for the detection of E. scophthalmi in infected farmed turbot. This methodology is applicable for routine controls on the farm at every stage of the parasite infection. Results of the study demonstrate the robustness, specificity, efficiency and reliability of the technique. In addition, this study also provides a non‐invasive procedure of sampling through swaps. This allows control, prevention and diagnosis of the parasite infection at turbot farms while maintaining the welfare of the cultivated fish and avoiding sacrifice of the fish sampled.  相似文献   

7.
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8.
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9.
Renibacterium salmoninarum is the causative agent of bacterial kidney disease, which significantly affects salmonid farming worldwide. Despite this impact, there is scarce data on its iron uptake ability, a factor of pathogenesis. This study investigated the iron acquisition mechanisms of R. salmoninarum and its capacity to uptake iron from different sources. Thirty‐two Chilean isolates and the DSM20767T type strain grew in the presence of 2,2′‐Dipyridyl at varying concentrations (250–330 μm ), and all isolates positively reacted on chrome azurol S agar. Subsequently, inocula of four Chilean isolates and the type strain were prepared with or without 200 μm of 2,2′‐Dipyridyl for uptake assays. Assay results revealed differences between the isolates in terms of iron acquisition. While a prior iron‐limited environment was, for most isolates, not required to activate the uptake of iron (II) sulphate, ammonium iron (III) citrate or iron (III) chloride at higher concentrations (100 μm ), it did facilitate growth at lower iron concentrations (10 μm and 1 μm ). An exception was the H‐2 isolate, which only grew with 100 μm of iron sulphide. In turn, 100 μm of haemin was toxic when isolates were grown in normal KDM‐2. In silico R. salmoninarumATCC 33209T genome analysis detected various genes coding iron uptake‐related proteins. This is the first study indicating two iron acquisition systems in R. salmoninarum: one involving siderophores and another involving haem group utilization. These data represent a first step towards fully elucidating this virulence factor in the pathogenic R. salmoninarum.  相似文献   

10.
Cathepsin L is one of the most important lysosomal cysteine proteases for the initiation of protein degradation, and it is involved in the immune response in many vertebrates. However, its function in the innate immune system of turtles remains poorly understood. Here, we cloned the cathepsin L gene from the Chinese soft‐shelled turtle Pelodiscus sinensis. We then examined the mRNA expression in different tissues and at different time points after infection by Aeromonas hydrophila or Vibrio parahemolyticus. The full‐length cDNA sequence cloned from P. sinensis was 1,805 bp with a 1,071 bp open reading frame, which encodes a 40.39 kDa polypeptide of 356 amino acids. The deduced protein sequence contained an active triad of Cys, His and Asn, and conserved ERWNIN and GNFD motifs. Homology analysis showed that the deduced amino acid sequence shared 77%–96% identity with other known species. Sequence alignment, phylogenetic analysis and structural comparisons revealed that cathepsin L is a member of the cathepsin family. Real‐time PCR analyses showed that turtle cathepsin L mRNA is ubiquitously expressed in various tissues, and the expression level is higher in the liver than in other tissues. The expression level in the liver peaks 24 hr after A. hydrophila infection and at two times (12 and 72 hr) after V. parahemolyticus infection. These results suggest that the cathepsin L gene of P. sinensis is likely involved in the process of the antibacterial response to A. hydrophila and V. parahemolyticus. This study is of great importance for future work exploring the molecular mechanism of antibacterial immune responses in P. sinensis.  相似文献   

11.
This study was conducted to investigate the effects of dietary β‐conglycinin on the growth performance, digestion, gut morphology and immune responses of juvenile turbot (Scophthalmus maximus L.). Four diets were formulated to contain 0%, 2%, 4% and 8% purified β‐conglycinin. Triplicate groups of 30 fish were fed to apparent satiation twice daily for 12 weeks. Fish fed 4% and 8% dietary β‐conglycinin showed significantly reduced specific growth rate, feed efficiency ratio, apparent digestibility coefficient of nutrients and whole‐body lipid contents, as well as a profound infiltration of mixed leucocytes in the lamina propria and a significant decrease in the absorptive surface of distal intestine. The expression of pro‐inflammatory cytokines, TNF‐α and IL‐1β, in the distal intestine was significantly upregulated by 4% dietary β‐conglycinin, whereas a significantly lower expression level of IgM and anti‐inflammatory cytokine TGF‐β1 was observed in fish fed 8% dietary β‐conglycinin. Serum lysozyme and alternative complement pathway activity were first significantly enhanced by 2% dietary β‐conglycinin and then rapidly declined by 4% and 8% dietary β‐conglycinin. Respiratory burst activity of head kidney macrophages and serum superoxide dismutase activity were significantly suppressed by 4% and 8% dietary β‐conglycinin. Dietary β‐conglycinin (2–8%) significantly increased the level of specific antibody against β‐conglycinin in serum. Collectively, these results suggested that higher levels of dietary β‐conglycinin (4–8%) induced a variety of non‐specific and specific immune responses and intestinal mucosal lesions in turbot, resulting in inferior feed utilization and poor growth performance.  相似文献   

12.
This study was aimed to find out the effect of five light spectra (white, LDW; blue, LDB; green, LDG; red, LDR and yellow, LDY) on the visual development and growth performance of turbot (Scophthalmus maximus) larvae. The number (per 100 μm) of cone cells, outer nuclear cells, ganglion cells, the ratio of outer nuclear to cone and ganglion cells, lens diameter and the minimum separable angle were determined. Variations in growth performance in total length, body mass and body colour were also examined. The results showed that the LDB group underwent metamorphosis faster than did the other groups and attained the greatest total length, body mass and light sensitivity. The LDB group also completed the body colour variation prior to the other groups. The LDG group exhibited a high mortality rate since 25 dph (days post‐hatching). We arrived at the conclusions that the spectra could affect the vision and body development of turbot larvae specifically and the blue light had the enhancing efficiency during this process, followed by the white and yellow light. The red light imposed an inhibiting effect on these variations. Our study highlighted the significance of light condition and provided the reference for the farming.  相似文献   

13.
The enteric myxozoan parasites Enteromyxum leei (Diamant, Lom et Dyková) and Enteromyxum scophthalmi Palenzuela, Redondo et Álvarez‐Pellitero are responsible for high weight loss in infected fish, which leads to subchronic disease and low mortality rates in gilthead sea bream (GSB), Sparus aurata L., and to high mortality rates in turbot, Psetta maxima (L.). The detection of initial parasite stages in histological sections is particularly difficult, but can be simplified by means of specific antibodies. Rabbit polyclonal antibodies (pAbs) were raised against E. scophthalmi and E. leei, and direct enzyme‐linked immunosorbent assay (ELISA) and immunohistochemistry were used to characterize their sensitivity and specificity. Both pAbs were adsorbed (apAb) with non‐infected intestines to avoid non‐specific labelling of fish tissues and to improve their specificity. The highest titre obtained in ELISA was 1: 32 000 for apAb‐Eleei and 1:16 000 for apAb‐Escoph. Working dilutions in immunohistochemistry were 1:1000 for apAb‐Eleei and 1:8000 for apAb‐Escoph. Both apAbs labelled proliferative and sporogonic stages with high specificity. apAb‐Escoph was very specific, whereas apAb‐Eleei cross‐reacted with Sphaerospora dicentrarchi Sitjà‐Bobadilla et Álvarez‐Pellitero and Sphaerospora testicularis Sitjà‐Bobadilla et Álvarez‐Pellitero, suggesting the presence of shared antigens. These pAbs stand as new tools for antigenic characterization and the diagnosis of both Enteromyxum species.  相似文献   

14.
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15.
A Thelohanellus species was encountered during a survey on Thelohanellus diversity of Carassius auratus gibelio (Bloch) in China. The infection is characterized by the presence of large cysts of 1.4–3.2 cm in diameter in the skin of host. Mature spores were ampullaceous in frontal view and testudinate in lateral view, measuring 19.7 ± 0.7 (18.6–20.8) μm long, 7.6 ± 0.4 (6.6–8.4) μm wide and 7.3 ± 0.5 (6.6–8.8) μm thick. The single polar capsule was elongated pyriform, with 11.1 ± 0.5 (10.0–11.9) μm long and 5.3 ± 0.3 (4.3–5.8) μm wide. Polar filaments coiled with 7–8 turns. Scanning electron microscopy revealed a smooth spore surface with flat side and convex side. The sutural line was straight or ‘S’ like, running near the middle of the valves. Histologically, the large cysts consisting of numerous small plasmodia developed in the dermis of the skin. The BLAST search indicated that the newly obtained ssrRNA gene sequences did not match any available sequences in GenBank and phylogenetic analysis placed it in the Thelohanellus clade. Based on morphology and molecular differences with reported Thelohanellus spp., this parasite was described as a new species of genus Thelohanellus.  相似文献   

16.
The geoduck clam, Panopea generosa, is a species from the west coast of Baja California, Mexico, and the optimization of seed production systems is still a limiting factor for its aquaculture. In this study, a flow‐through culture system was designed and tested in P. generosa larvae. Survival and growth was compared in triplicate 45‐L fiberglass tanks using three larval densities (5, 10 and 15 larvae/ml). A head tank kept constant the water inflow, with a daily renewal rate of 1.8× tank volume. The food (Isochrysis spp.) was dosed according to the ingestion rate of larvae and the dilution rate. Survival decreased linearly during the first 10 days and reached asymptotic values of ca. 20% (15 larvae/ml) and 50% (densities of 5 and 10 larvae/ml) afterwards. Mean shell length at the end of the experiment (243 ± 1.8 to 270 ± 0.7 μm) was not statistically different among treatments, even though a trend towards higher gross growth rate was observed in the treatment with the lowest density (9.5 μm/day) relative to the rest of the treatments (8.5 μm/day). It is concluded that P. generosa larvae can be successfully grown in flow‐through systems at maximum densities of 10 larvae/ml without significantly affecting their survival and growth rates. The system design was reliable, kept a constant water flow with reduced maintenance, and may represent an important option in the laboratory for increasing the stocking density of Panopea species during the larval phase.  相似文献   

17.
The nematode Panagrolaimus sp. was tested as live feed to replace Artemia nauplii during first larval stages of whiteleg shrimp Litopenaeus vannamei. In Trial 1, shrimp larvae were fed one of four diets from Zoea 2 to Postlarva 1 (PL1): (A) Artemia nauplii, control treatment; (NC) nematodes enriched in docosahexaenoic acid (DHA) provided by the dinoflagellate Crypthecodinium cohnii; (N) non‐enriched nematodes; and (Algae) a mixture of microalgae supplemented in C. cohnii cells. In Trial 2, shrimp were fed (A), (NC) and a different treatment (NS) with nematodes enriched in polyunsaturated fatty acids (PUFAs) provided by the commercial product S.presso®, until Postlarva 6 (PL6). Mysis 1 larvae fed nematodes of the three dietary treatments were 300 μm longer (3.2 ± 0.3 mm) than control larvae. At PL1, control shrimp were 300 μm longer (4.5 ± 0.3 mm) than those fed DHA‐enriched or PUFAs‐enriched nematodes. No differences were observed in length and survival at PL6 between control larvae and those fed DHA‐enriched nematodes (5.1 ± 0.5 mm; 33.1%–44.4%). Shrimp fed microalgae showed a delay in development at PL1. This work is the first demonstration of Panagrolaimus sp. suitability as a complete substitute for Artemia in rearing shrimp from Zoea 2 to PL6.  相似文献   

18.
As one of the most serious pathogens in mariculture, the outbreaks of grouper Iridovirus (SGIV‐Gx) caused high mortality rates in cultured groupers in Guangxi, China. Hence, effective medicines for fighting against grouper Iridovirus are urgently needed. The possible application of Curcuma kwangsiensis ingredients against SGIV‐Gx infection was evaluated in vitro and in vivo in this study. The safe working concentration of each C. kwangsiensis ingredient was identified (C. kwangsiensis ethanol ingredient, CKEE ≤5 mg/ml; curcumin ≤20 μg/ml; curdione ≤500 μg/ml; curcumenol ≤500 μg/ml; curcumol displayed no cytotoxic effects even at 2 mg/ml) in vitro and in vivo. The inhibitory activities of each C. kwangsiensis ingredient against SGIV‐Gx infection were analysed using aptamer (Q2)‐based fluorescent molecular probe (Q2‐AFMP) and RT‐qPCR. The results showed that C. kwangsiensis ingredient (CKEE, curcumin, curcumol, curdione and curcumenol) displayed antiviral activities against SGIV‐Gx infection in a dose‐dependent manner. Furthermore, according to the inhibitory percentage analysed using RT‐qPCR results, CKEE and curdione had the best antiviral activity against SGIV‐Gx above 93%. Overall, the results suggest that some C. kwangsiensis ingredients have excellent antiviral effects, making it an interesting candidate for developing effective medicines for preventing and controlling SGIV‐Gx outbreaks in mariculture.  相似文献   

19.
Samples of invertebrates, primary producers, seawater and cores were taken surrounding several aquaculture sites in coastal British Columbia, Canada. Analysis of dissolved organic carbon (DOC) showed a significant decrease (P = 0.027) over 1 km and an overall average of 290 ± 40 μM at 1 m depth. At 10 m, the decrease was not significant and DOC averaged 81 ± 3 μM. The essential fatty acid 22:6ω3 (docosahexaenoic acid, DHA) was significantly lower nearer to the farm sites for all mollusc taxa as well as for mussels alone (P < 0.01). However, mussel wet weight as well as the zooplankton fatty acid marker (ZFA; Σ 20:1ω9, 22:1ω11, 22:1ω9) and 20:1ω9 individually, decreased significantly with distance from the farms (P < 0.03). Principal components analysis showed a similar trend with DHA being higher in molluscs further from the farm. Mussels had significantly higher amounts of DHA compared with other molluscs indicating their potential in multi‐trophic, co‐culturing systems. Bacterial fatty acid markers increased in molluscs with distance from the farms; however, 18:1ω7, which is representative of methane‐oxidizing bacteria, remained higher closer to the farm. In addition, DHA proportions in mussels showed a significant breakpoint at 339 m from the farm suggesting co‐cultured mussels would benefit from placement further than 339 m from farms where DHA would be optimized.  相似文献   

20.
Four experimental diets were fed to turbot to examine the effect of fish hydrolysate and ultra‐filtered fish hydrolysate on growth performance, feed utilization and non‐specific immune response. Fish hydrolysate was produced by enzymatic treatment and size fractionated using ultra‐filtration (UF). The permeate (molecular weight <1000 Da) after UF and the non‐ultra‐filtered fish hydrolysate (NUF) were tested as feed ingredients. Diets UF1, UF2 contained 3.7%, 1.2% ultra‐filtered fish hydrolysate to replace fish meal protein respectively. The diets UF1, NUF were identical in composition except that the molecular weight of fish hydrolysate in the diet. Fish meal was used in the control diet. All diets were made equal in protein, lipid and energy. Each experimental diet was fed to juvenile turbot (27.87 ± 0.04 g) in triplicate for 8 weeks. Results of this study indicate that the best overall growth and feed utilization of turbot juveniles were obtained with a diet containing higher dose of the small molecular weight compounds in fish hydrolysate. Acid phosphatase, alkaline phosphatase, lysozyme and superoxide dismutase activity in serum were not affected by diet. Total antioxidant capacity was improved with increasing level of low molecule weight fish hydrolysate (UF1).  相似文献   

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