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1.
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2.
EmpA is an extracellular metalloprotease secreted by Vibrio anguillarum.For better understanding its role in the patho-genicity of V.anguillarum strain M3,empA insertion mutant was constructed.In the mutant it decreased in extracellular proteolytic activity,swarming motility,hemolytic activity and virulence on turbot(Scophthalmus maximus).Significant decline(by 5-fold)of extracellular proteolytic activity and similar growth curve between mutant and wild type strains indicated that EmpA was the major extracellular protease of M3.LD50 of mutant increased by 38-fold compared with wild type.No pro-EmpA was detected in the su-pernatant of culture,indicating that EmpA autolyzed to mature protein after 24 h.Secretion of EmpA in M3 was similar to that in NB10 strain.Attenuated virulence of mutant was similar to that of M93Sm strain.It was demonstrated that specific operation of EmpA was different from that in previous studies and EmpA contributed to the swarming motility and hemolytic activity in V.an-guillarum strain M3.The results provides insight into understanding the function of EmpA and its potential application in vaccine development.  相似文献   

3.
Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification (LAMP) assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase (empA) gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65°C in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V. anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field.  相似文献   

4.
Vibrio anguillarum is a common bacterial pathogen in fish.However,little is known about its pathogenic mechanism,in part,because the entire genome has not been completely sequenced.We constructed a fosmid library for V.anguillarum containing 960 clones with an average insert size of 37.7 kb and 8.6-fold genome coverage.We characterized the library by end-sequencing 50 randomly selected clones.This generated 93 sequences with a total length of 57 485 bp covering 1.4% of the whole genome.Of these sequences,58...  相似文献   

5.
Freeze-drying, continuous passage and ultra-low temperature cryopreservation are often used to preserve pathogens. In this study, Vibrio anguillarum was rejuvenated by intramuscular infection as the initial strain. The difference between cells preserved with different preservation methods and their initial strains were compared with physiological and biochemical methods and through antibiotics resistance analysis. The composition of protectants for freeze-drying V. anguillarum was optimized. We found that the optimal composition of protectants was 8% of trehalose, 12% of skim milk, 8.0% of lactose, 2.0% of sodium citrate, 12.0% of serum and 8.0% of mannitol. The indexes of lysine decarboxylase and urease changed after continuous passage. The urease reaction changed after freeze-drying and freeze-thawing, but the reaction can be restored to the initial after freeze-drying. Based on the antibiotics resistance analyses, the sensitivity of V. anguillarum to different drugs including rifampicin, erythrocin, furazolidone, ceftazidime, lomefloxacin, gentamycin, azithromycin, doxycycline, ampicillin, co-trimoxazole and cefoperazone changed after different treatments, and some of these changes can be restored to the original through activation culture. In sum, compared with cryopreservation and continuous passage, the freeze-drying is more sustainable for the long-term preservation of V. anguillarum, which showed a better effect in maintaining the original characteristics of pathogen.  相似文献   

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7.
Interleukin-17(IL-17), a prototype member of IL-17 family, plays an important role in defending against extracellular pathogens as a pro-inflammatory cytokine. The function and distribution of IL-17 have been extensively studied in many vertebrates. However, few study has focused on the role of IL-17 in invertebrates, especially in mollusks. In this study, an IL-17 homolog was identified in Octopus ocellatus, which was designated as OoIL-17. The phylogenetic analysis showed that OoIL-17 is clustered well with other invertebrate IL-17, indicating it is highly similar with the IL-17 of other invertebrates. The expression of OoIL-17 gene was analyzed with qRT-PCR in a variety of healthy tissues and the hemocytes infected with Vibro anguillarum or Micrococcus luteus. The mRNA of OoIL-17 gene is constitutively expressed at different levels in all examined tissues of healthy O. ocellatus, including mantle, stomach, hemocytes, muscle, gonad, hepatopancreas, systemic heart and gill. The lowest expression was observed in mantle while the highest was observed in hepatopancreas. The expression level of OoIL-17 gene is significantly up-regulated in O. ocellatus hemocytes upon infection with V. anguillarum and M. luteus, indicating its active involvement in the host immune response against bacterial pathogens. The results laid the foundation for further understanding the innate immune mechanisms of IL-17 in O. ocellatus and mollusks.  相似文献   

8.
Gao  Xinming  Yang  Haiyan  Tang  Daojun  Du  Chen  Jin  Shan  Hou  Congcong  Zhang  Chundan  Zhu  Junquan  Wang  Jianping 《中国海洋湖沼学报》2022,40(2):643-655
Journal of Oceanology and Limnology - Temperature is an important factor affecting the growth, development, and survival of organisms. The effects of temperature stress on aquatic organisms have...  相似文献   

9.
We cloned and sequenced a prtV-like gene from Vibrio anguillarum M3 strain.This prtV gene encodes a putative protein of 918 amino acids,and is highly homologous to the V.cholerae prtV gene.We found that a prtV insertion mutant strain displayed lower gelatinase activity on gelatin agar,lower protease activity against azocasein,and lower activity for four glycosidases.This prtV mutant strain also had increased activity for two esterases in its extracellular products,as analyzed by the API ZYM system.In additi...  相似文献   

10.
DNA damage of aquatic organisms living in polluted environments can be used as a biomarker of the genotoxicity of toxic agents to organisms. This technique has been playing an important role in ecotoxicologlcal study and environmental risk assessment. In this article, main types of DNA damage caused by pollutants in water environments were reviewed; methods of detecting DNA damage were also documented for water environmental monitoring.  相似文献   

11.
Ammonium overloading is a common response of aquatic organisms to air exposure during transport. This study elucidated the relationship between ammonium overloading and mortality of crab Portunus trituberculatus. Additionally, we also explored the effects of emersion time and air temperature on ammonium loading and concomitant physiological change. To test the air temperature effect, the crab was exposed to 16, 23 and 30℃ in air for 3 h, respectively, and then recovered in seawater at 23℃ for 12 h. To test the emersion time effect, crab was exposed to 23℃ in air for 0.5 and 3 h, respectively, and then recovered in seawater at 23℃ for 12 h. In the control group, crab was always immersed at 23℃. At each time interval(0.5, 1.5 and 3 h during air exposure and 0.5, 2, 4 and 12 h during recovery), ammonium excretion rate, level of total ammonium, total free amino acids and urea concentration in hemolymph and the hepatopancreas enzyme activity involved in detoxifying ammonium were analysed. Results showed that crab mortality was positively related with emersion time and temperature while ammonium loading was lower at 16 and 30℃ than at 23℃. For crab experiencing thermal inconsistence of culture media(i.e., 16 or 30℃), they were higher in ammonium excretion rate and activities of ammonium detoxification enzymes, which may be the reason that they had a lower ammonium loading. Prolonged emersion time(3.0 h vs. 0.5 h) increased the ammonium overloading and the activity of ammonium detoxification pathways in crab. Our results demonstrated that emersion-induced ammonium overloading may not be the main reason leading to P. trituberculatus death during air exposure and subsequent recovery. When the culture medium changed, thermal variation, compared with constant temperature, could reduce ammonium overloading in crab by elevating the activities of ammonium detoxification enzymes and ammonium excretion rate during recovery period.  相似文献   

12.
Mutant J61321 with enhanced siderophore production of Alteromonas aurantia AI8 was obtained after a series of chemical-physical mutageneses. It was found that the antibacterial activity against Vibrio anguillarum W-1 and siderophore production of the mutant were higher than those by the original strain A 18 which had been used in mariculture. The results of the specific assay（Csaky and Arnow methods） of siderophore showed that the sidrophore with hydroxamate group was produced by mutant J61321 and the original strain A 18, respectively, while the siderophore with catechol group was yielded by strain W-1 （Aibrio anguillarum）. Meanwhile, the siderophore yield, antibacterial activity and anti-chelator activity of strain J61321 were higher than those of its parent strain A 18.  相似文献   

13.
Antibiotics are widespread in various environmental media,and may pose a potential threat to aquatic ecosystems and non-target aquatic organisms.Florfenicol(FLO) is one of the most commonly used antibiotics in aquaculture,and extensively used to substitute chloramphenicol with its strong sterilization and low adverse ef fect.In this study,flounder Paralichthys olivaceus,an important economic fish species in seawater was used as an experimental subject.Five exposure concentrations of FLO(including environmentrelated concentrations) were set at 0,0.01,0.1,1,and 10 mg/L.Ef fects of FLO exposure for 168 h on growth and development,motor behavior,antioxidant enzyme activity,malondialdehyde(MDA) content,and thyroid hormone level of P.olivaceus larvae were studied in pre-larvae(1 dpf) and post-larvae(20 dpf).The results show that the short-term FLO exposure could promote the larvae growth to some degrees,but inhibit them as the exposure time prolonged.For pre-larvae,FLO at 0.01 mg/L could stimulate the motor nerve system and increase the swimming ability,but inhibited it at 1 mg/L.With the increasing dosage of FLO,the superoxide dismutase(SOD) and MDA contents were elevated,reaching the maximum in the 1 mg/L FLO group.The pre-larvae were more sensitive than the post-larvae to FLO in the environment,and the growth and immune resistance could be damaged with long exposure.Post-larvae were more tolerant to external pollutants,FLO at 1 mg/L could promote the motor behavior and reduce SOD and MDA contents.Therefore,FLO can be used as an antibiotic at a proper concentration but as a drug to prevent disease in a long-term way.  相似文献   

14.
This article presents the results ofhydrochemical and hydrobiological studies of shallow saline lakes of the Borzya group （Zabaikalsky K_rai, Russia） at the initial filling phase. The lake-margin ecosystems of the studied lakes are characterized by varying degrees of salinity from polyhaline to brine water. Cyclical variations of meteorological conditions and high salinity determined that the compositions of the aquatic organisms are specific, mostly between stenohaline and euryhaline species, the quantities are low, and the community structures are simple.  相似文献   

15.
Plant biomarkers, such as hydrocarbon waxes, are frequently found in various sediments and could be adopted as paleovegetation and paleoclimate indicators. Nevertheless, scarce researches have focused on leaf waxes in higher plants of alpine region.Herein, hydrocarbon leaf wax components of Salix oritrepha, which flourish in Nianbaoyeze Mountains in eastern Tibetan Plateau were fully discussed. The n-alkane distribution in leaves ranges from n-C_(21) to nC_(29) with maxima at n-C_(25), which were entirely different with Salix taxa displayed in previous surveys in non-alpine regions. The unusual even carbon nalkenes from n-C_(22:1) to n-C_(30:1), which were thought to appear only in aquatic organisms, were firstly reported in an alpine plant. Additionally, iso-(2-methyl) alkanes, ranging from i-C_(23) to i-C_(29) with maxima at i-C_(25), which have been commonly reported in microorganisms, were also identified in an alpine plant for the first time. Unusual hydrocarbon distribution detected in Salix oritrepha leaf from Nianbaoyeze Mountains is most likely due to the extreme environment in such alpine region.  相似文献   

16.
Bacteria-infected turbots Scophthalmus maximus with septicaemia were examined between 2001 and 2004 in aspects of the conditions of disease occurrence, clinical syndromes and pathological changes. The phenotypic information of pathogenic bacteria was studied, including morphology, physiological and biochemical characteristics, and the mol% G C of the DNA. In addition, representative strains (S010623-1, LH031120-1) were selected for molecular identification by partial 16S rRNA gene sequencing. The results show that the isolates (LH031120-1 to LH031120-6, HT040308-1 to HT040308-6, HT040620-1 to HT040620-6) from three farms were identified as Edwardsiella tarda. The isolates (S010610-1 to S010610-10, S010623-1 to S010623-20) from one farm were identified as Listonella anguillarum. We conducted studies on the pathogenicity of isolates by artificial infection, and revealed all infected groups in morbidity and mortality. The septicaemia infected turbot showed a syndrome similar to that of the naturally infected fish. Antibiotic sensitivity showed that of 37 antimicrobial agents, E. tarda was sensitive to 27 agents, and L. anguillarum was sensitive to 21 agents.  相似文献   

17.
Stable isotope methods can be used to determine the food sources and prey items of aquatic organisms accurately andreliably.This study examined the relative contribution of artificial foods (the formulated feed and Artemia) and natural foods toshrimp growth in an intensive Fenneropenaeus chinensis pond by using carbon and nitrogen stable isotopes.The results showed that the nutrition utilization efficiency of the harvested shrimp was low,only 33.18% of feed nitrogen and 21.73% of feed carbon beingconverted to shrimp flesh.Our stable isotope results showed that the shrimp obtained nutrition for maximum growth from artificialfoods,whose contribution was 93.5%,with the remaining attributed to the natural foods.However,there was 0.94 t harvested shrimpderived from natural foods (the rest of 13.56t harvested shrimp derived from artificial foods) in lha intensive pond with a shrimpproduction of 14.50 tha-1.Therefore,unit area shrimp production can be increased by increasing the contribution proportion of natu-ral foods in intensive shrimp farming.  相似文献   

18.
Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD50 value of 1.0×105 colony forming units (CFU)mL-1, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillarum with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay (IFA) was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature.  相似文献   

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20.
Chao  Jinyu  Feng  Song  Hao  Yingdong  Lin  Jianing  Zhang  Bin 《中国海洋湖沼学报》2023,41(1):251-266

To date, little attention has been paid to the effects of organophosphate esters (OPEs) pollution on aquacultural environment and aquatic product safety. Huanghe (Yellow) River delta area is one of the largest aquaculture centers in China, where ecological security protection is crucial in the national strategy of China. To explore the pollution characteristics, bioaccumulation, and health risks of OPEs in aquaculture farms in the Huanghe River delta and natural water bodies in the adjacent seas, five species of organisms from different farm types nearby the Huanghe River delta, and the corresponding culture water and sediments were sampled in this study. The total concentrations of ∑13OPEs in water, sediments, and organisms were 51.53–272.18 ng/L, 52.63–63.17 ng/g dry weight (dw), and 46.82–108.90 ng/g dw, respectively. Among the five types of culture ponds, the water samples from the swimming crab and hairy crab culture ponds exhibited higher OPEs, the concentration of OPEs in the sediments from the few ponds was relatively balanced, and the OPEs in the organism from the holothurian ponds was higher. Tris (1,3-dichloro-2-isopropyl phosphate) (TDCP) was the main contaminant in water samples and tripropyl phosphate (TPrP) in sediments and organisms. However, trisphenyl phosphate (TPhP) showed the strongest bioaccumulation ability, followed by 2-ethylhexyl diphenyl phosphate (EHDPP) and TPrP. The bioaccumulation capacities of the five species were as follows: prawn > holothurian > hairy crab > swimming crab > carp. These five types of organisms, as main seafood in human consumption, were at low risk of negative impacts of pollution. However, the risk from the mixture of organophosphate flame retardants (OPFRs) still requires more attention due to the increasing consumption and production in the world.

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