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1.
Bacteria isolated from an outbreak with moderate mortalities in farmed sole, Solea senegalensis (Kaup), in the south of Spain were identified as Vibrio harveyi and V. parahaemolyticus. Only bacterial strains showing swarming were virulent in sole and caused mortalities in experimentally inoculated fish. However, the signs of the disease were only reproduced with V. harveyi. The intramuscular inoculation of the extracellular products (ECPs) of both species produced mortalities in inoculated fish and the appearance of surface ulcers in the case of V. harveyi. However, the inoculation of sublethal doses of ECPs to fish showed a protective effect against V. harveyi.  相似文献   

2.
ABSTRACT:   Among several marine antagonistic bacteria isolated from the sea water of Kagoshima Bay, Pseudoalteromonas sp. A1-J11 was found to produce anti- Vibrio substances. The anti- Vibrio substances were extracted from the culture supernatant of the strain with chloroform and isolated using reverse-phase (Cosmosil 75C18-OPN) column chromatography followed by high-pressure liquid chromatography (Mightysil RP-18 GP Aqua). Purified substances, designated as AVS-03 a , c and d showed similar ultraviolet absorption spectra with λmax at 215, 235, 315 and 327 nm in methanol. AVS-03 d , the major anti- Vibrio substance, was thermostable up to 100°C and pH stable over a pH range higher than 4.0, and also showed strong inhibitory activities, specifically against Vibrio harveyi strains.  相似文献   

3.
本研究以引起珍珠龙胆石斑鱼[Epinephelus fuscoguttatus(♀)×Epinephelus lanceolatus(♂)]幼鱼“皮肤溃疡病”的哈维氏弧菌(Vibrio harveyi) ML01株为研究对象,采用平板膜覆盖技术和柱层析技术,分离、纯化了ML01株的胞外产物及分泌性蛋白.应用毒性试验、质谱分析与分子克隆技术,对纯化的胞外产物和3种主要的分泌性蛋白进行了特性分析与鉴定.结果显示,哈维氏弧菌ML01株的胞外产物(Extracellular products,ECPs)具有酯酶、明胶酶、淀粉酶、酪蛋白酶活性,无脲酶活性.ECPs对羊红细胞无溶血性,对斑马鱼(Danio rerio)的半数致死剂量(LD50)为19.55μg/g鱼体重.从ML01株中分离到3种主要的分泌蛋白P42、P36、P31,其分子量分别为42、36、31 kDa.经质谱鉴定和分析,这3种蛋白分别为哈维氏弧菌的外膜蛋白OmpU和OmpN,以及一种功能未知的蛋白.利用同源克隆,成功地从ML01株基因组中扩增到了P42、P36、P31的基因.序列测定和比对结果显示,ML01株的这3个基因与哈维氏弧菌ATCC 33843(GenBank CP009467)的相应基因相比,其开放阅读框(Open reading frame,ORF)序列的相似性分别为97.08％、100％、99.67％,其编码的多肽序列的相似性分别为99.71％、100％和99.93％.本研究对进一步分析哈维氏弧菌ML01株的致病机理、研发该菌的亚单位疫苗具有重要的参考价值.  相似文献   

4.
Bacterial subcellular components and probiotics were successful for the stimulation of immunity and the prevention of Vibrio harveyi infections in rainbow trout, Oncorhynchus mykiss (Walbaum). Rainbow trout were immunized with whole inactivated cells of V. harveyi to obtain polyclonal antibodies against specific antigens. Western blotting showed a unique reactive band (∼93 kDa) between serum and bacterial proteins from outer membrane proteins (OMP) and extracellular products (ECP). Probiotics were selected according to their capability to inhibit V. harveyi . Two of these bacteria, i.e. A3-47 and A3-51, showed cross-reactivity with V. harveyi antiserum. Their OMPs and ECPs were reactive with V. harveyi antiserum in bands of ∼93 kDa for A3-51 and higher for A3-47. In vivo tests determined that fish fed with A3-51 produced cross-reactive antibodies against V. harveyi and also, the survival of these fish infected with V. harveyi was high, being similar to the level achieved with vaccinated fish. Thus, the probiotics, when administered as live preparations, were capable of producing cross-reactive antibody against specific bacterial pathogens.  相似文献   

5.
Mycobacterium spp. isolated from food and ornamental fish in Thailand (TB1, TB40, TB267 and TB268), and the type strains Mycobacterium marinum (NCIMB 1298), M. fortuitum (NCIMB 1294) and M. chelonae (NCIMB 1474) were cultured in Long's medium, Eagle's minimum essential medium, Sauton's medium and modified Sauton's medium. The latter enabled excellent growth and production of extracellular products (ECP) from TB 40, TB267, TB268 and M. marinum NCIMB 1298 in particular, whereas growth and production of ECP for all strains was limited in Long's medium. SDS-PAGE protein profiles of ECPs from 14-day culture supernatants showed major bands at 65 and <14 kDa. After 2 days culture at the higher temperature of 37°C (heat shock), the production of ECP from all mycobacteria strains except M. marinum averaged approximately four- to 10-fold higher than from strains cultured for 14 days at 28°C. Enzyme testing for the type strains indicated only mucinase activity for M. marinum, while lipase and RNase activities were detected for M. chelonae and M. fortuitum . Protease and DNase activities could not be detected for any of the Mycobacterium spp. tested. The medium lethal dose (LD50) of ECP to rainbow trout and Nile tilapia was greater than 400 μg protein fish–1.  相似文献   

6.
Abstract Yellowfin sea bream Acanthopagrus latus (Houttuyn) (0.98 ± 0.27 g) were examined for temperature and salinity tolerances at nine different environmental regimes (0.3, 15 and 33 ppt combined with 10, 25 and 32 °C), in which the fish were subjected to gradual and sudden changes in temperature and salinity respectively. The 50% CTMax (critical thermal maximum) and the UILT (upper incipient lethal temperature) were in the range of 34.8–38.2 °C and 32.8–36.4 °C respectively. The 50% CTMin (critical thermal minimum) and the LILT (lower incipient lethal temperature) were in the ranges of 4.9–9.4 °C and 7.1–17.6 °C respectively. The 50% CSMax (critical salinity maximum) and the UILS (upper incipient lethal salinity) were in the ranges of 54–69‰ and 9–44‰, respectively. The fish at lower temperature (10 °C) and salinity of 33‰ tolerated temperature as low as 6 °C when the temperature was decreased gradually. The fish at 33 and 15‰ and lower temperature (10 °C) tolerated salinity as high as 66–67‰, whereas those at freshwater salinity level (0.3‰) and 32 °C tolerated salinity to 50‰ when salinity was increased gradually. The relationships among UILT, LILT, 50% CTMax, 50% CTMin, UILS, 50% CSMax, salinity and temperature are given.  相似文献   

7.
ABSTRACT:   The effects of temperature changes on the endogenous rhythm of oxygen consumption in the Japanese flounder Paralichthys olivaceus which were acclimated to 3, 14 or 20°C in advance were investigated. The oxygen consumption rate (OCR) of individuals (total length >28 cm) that were acclimated to 3°C, maintained a relatively low value of 6.9 ± 2.3 mL (mean ± SD) O2/kg ww (wet weight)/h till 5.8°C, which is considered a 'minimum' rate. Thereafter, the OCR gradually increased up to 18.7 mL O2/kg ww/h at the temperature of 10.6°C. The OCR of individuals acclimated to 14°C stabilized with 124.2 ± 24.1 mL O2/kg ww/h, when tested at the relatively constant temperature of 14.4 ± 0.17°C. Maximum entropy spectral analysis revealed that the peak OCR occurred at 23.9 h intervals, which could be a circadian rhythm. The mean OCR showed a peak at 06.00–08.00 hours. For the temperature above 25°C in which the fish were acclimated to 20°C, the rhythmic patterns of OCR were observed until the experimental temperature reached 26.4°C. For the temperature above 28.4°C, the metabolic activity rhythm of the OCR was dampened and then the OCR abruptly rose again. It is assumed that the oscillator of the endogenous rhythm lost the governing of normal metabolic activity in Japanese flounders above 28.4°C. These results suggest that the temperatures of around 5.8, 26.4 and 28.4°C are the 'lower limit temperature', 'upper incipient lethal temperature', and the 'critical thermal maximum', respectively, for the endogenous rhythm of oxygen consumption in the Japanese flounder.  相似文献   

8.
ABSTRACT:   Fast skeletal muscles of Japanese flounder Paralichthys olivaceus and red sea bream Pagrus major were examined for quantitative and qualitative changes of mitochondrial ATP synthase (FoF1-ATPase) in association with rearing temperatures. The specific activities of FoF1-ATPase from Japanese flounder reared at 10°C, 15°C and 25°C for 4 weeks were determined to be 81 ± 11, 74 ± 13 and 83 ± 11 nmol/min·mg mitochondrial protein, respectively. The corresponding activity from red sea bream reared at 8°C for 5 weeks was determined to be 65 ± 9 nmol/min·mg mitochondrial protein, which was higher than 33 ± 9 nmol/min·mg mitochondrial protein in fish reared at 23°C. The contents of α- and β-F1-ATPase in total mitochondrial proteins were not significantly different between fish reared at different temperatures for the two fish species. However, the contents of β-F1-ATPase in the total fast skeletal muscle extracts, prepared from Japanese flounder reared at 10°C, were 2.1- and 2.9-fold higher than those for fish reared at 15°C and 25°C, respectively. The corresponding content from red seabream reared at 8°C was 2.2-fold higher than that for fish reared at 23°C. Therefore, the changes in FoF1-ATPase depending on rearing temperatures were species-specific.  相似文献   

9.
We evaluated the toxicity of commercial injectable erythromycin (Erythro-200) and the injectable drug carrier alone (PEG-400, ethyl alcohol and ethyl acetate) in sub-adult spring chinook salmon, Oncorhynchus tshawytscha (Walbaum). Both compounds were toxic to salmon when administered in high dosages. The 50% lethal dose (LD50) of the erythromycin within its carrier (Erythro-200) at 12 °C was estimated at 429 mg erythromycin kg–1 or 2.145 mL kg–1 when measured by volume. The LD50 of the drug carrier alone was 2.95 mL kg–1 by volume or equivalent to 3.21 g kg–1 of the carrier. We evaluated and scored the histopathology in kidney sections removed from fish 96 h after injection with Erythro-200, the drug carrier alone, or sterile phosphate buffered saline (PBS) that served as a reference control. Kidneys from fish injected with saline had lowest average pathology scores; those injected with the carrier had moderate scores; and the pathology score was highest in tissues from erythromycin-injected fish. In tests at 12 °C, vacuoles were more prominent in proximal and distal tubules of fish injected with higher dosages of 403 and 545 mg erythromycin kg–1 than in tissues from fish injected with 299 mg kg–1.  相似文献   

10.
ABSTRACT:   Tropical minute rotifer strains (SS-type) induce mixis at 30–35°C but sexual reproduction and resting egg formation do not proceed well due to rapid environmental change. The present study examined the effect of temperature regulation on rotifer Brachionus rotundiformis (Langkawi strain, SS-type) resting egg formation in small (500 mL in culture volume)- and large-scale (500 L in culture volume) experiments. Rotifers were cultured at 30°C in 15–17 p.p.t. seawater with an initial density of 1 individual (ind.)/mL. After 4 days, when cultures were in exponential growth stage with active mixis induction, the culture temperature of the experimental rotifers was changed to 25°C. Control rotifers were cultured at 30°C throughout the experiment. Fresh or frozen Nannochloropsis oculata and condensed freshwater Chlorella vulgaris were used as the rotifer diets in the small- and large-scale experiments, respectively. Significantly higher resting egg production was observed with the experimental rotifers (30→ 25°C) versus the control rotifers. In the large-scale trial, experimental rotifers produced 2.6 × 106 resting eggs during a 9-day experiment, which was 1.6-fold more than the control rotifers. Moreover, the efficiency of resting egg formation was found to increase by a factor of 1.8. The present study indicates that decreasing culture temperature from 30 to 25°C after active mixis increased resting egg formation in B. rotundiformis (SS-type).  相似文献   

11.
ABSTRACT:   A rapid clustering scheme for Vibrio specieswas developed based on the analysis of 16S rDNA, which compriseda group-specific polymerase chain reaction (PCR) and restriction fragmentlength polymorphisms (RFLP) of the PCR-amplified 16S rDNA. The group-specificPCR, using a specific primer Vib1F, clustered the Vibrio speciesinto two large groups: Vib1F+ group and Vib1F– group.The PCR-RFLP, using Sca I and Bln I, further dividedthese groups into smaller groups. Finally, 46 Vibrio speciesand eight related species could be clustered into 14 groups usingthis rapid clustering method. Seasonal dynamics of the Vibrio communityin Yoshimi Bay, Hibiki-nada Sea, Japan, were examined based on therapid clustering method. In both seawater and sediments, the groupcomprised Vibrio parahaemolyticus , Vibrio alginolyticus , Vibriocampbellii , Vibrio carchariae , Vibrio harveyi and Vibrionatriegens and was predominant when the seawater temperaturewas above 20°C, whereas the group of Vibrio splendidus biotypeI and Vibrio lentus was abundant when the temperature wasaround or below 20°C.  相似文献   

12.
The growth and mortality of Neptunea arthritica juveniles hatched from different egg masses (controlled conditions and field) reared at 10 and 15 °C, and provided with sardine (D1) and polychaete (D2) as foods were evaluated. Results showed that diet type and water temperature significantly affect the growth and mortality. Interaction between these factors reflected robust relationship among them, but restricted exclusively to growth. Through the study period, juveniles treated at 10 °C showed maximum increments of 2.5 mm (shell length), 2.3 mm (shell width) and 0.25 g (body weight) from their initial size. While maximum values of those treated at 15 °C for the above-mentioned parameters were 7.6 mm, 5.5 mm and 1 g respectively. In both cases, these increments were recorded for individuals fed with sardine. Mortality was significantly affected by diets and water temperature, which significantly increased under D2 [10 °C (20–67%), 15 °C (70–87%)] or 15 °C [D1 (30–83%), D2 (70–87%)]. According to the results, juveniles of N. arthritica could be maintained under controlled conditions, which allows growth improvement. Mortality appears to be the limiting factor, but this could be improved by the implementation of efficient culture method, basically oriented to increase the water quality.  相似文献   

13.
ABSTRACT:   Feeding activity was examined at high temperatures by using a demand feeder for thermally selected and normal rainbow trout Oncorhynchus mykiss strains. When the water temperature was raised in experiment 1 from 17.5 to 25.7°C at 0.3°C/day in 21 days, the average daily food consumption rate in the thermally selected strain rose to 7.1%, which was significantly higher than that of the normal strain (4.1%, P  < 0.05). The corresponding rate was also significantly higher in the thermally selected (0.8%) than in the normal strain (0.2%, P  < 0.05) of fish in experiment 2 where water temperature was raised to 24.1°C in 0.5°C/day increments. When water temperature was raised rapidly in experiment 3 from 16.7 to 21.7°C in one day and gradually to 24.4°C in 28 days at 0.1°C/day, the average daily food consumption rates were 1.0 and 0.1% for the thermally selected and normal strains, respectively, with significant difference ( P  < 0.01). These results suggest that the thermally selected strain has acquired thermal tolerance as a result of artificial selection.  相似文献   

14.
The effects of temperature on growth, food-conversion ratio (FCR) and feeding intensity of juvenile dusky kob Argyrosomus japonicus (23.7±2.6 g fish−1) were assessed over the temperature range 17.5–28.5 °C in a 42-day growth trial. Growth increased with increasing temperature up to an optimum after which it declined. Specific growth rates were 2.05% and 1.2% day−1 for the fastest (25.3 °C) and the slowest (17.5 °C) treatments respectively. Food-conversion ratio peaked at a lower temperature than growth. Optimal (0.72 kg kg gain−1) and least efficient (1.40 kg kg gain−1) FCR were found at 21.7 and 17.5 °C respectively. Feeding intensity was linearly related to temperature within the range of 17.5–28.5 °C. These results corresponded to the thermal preference (25–26.4 °C) and natural temperature distribution (12–28 °C) of South African dusky kob. Determination of the temperature range that does not limit growth is a prerequisite to assess the relationship between growth and environmental variables such as light intensity, feeding regime and stocking density. Consequently, experiments to determine the effects of these environmental variables on growth and aquaculture potential of dusky kob should be conducted at 24–26 °C.  相似文献   

15.
Abstract –  The reproductive activities of two dominant fish species, roach Rutilus rutilus and rudd Scardinius erythrophthalmus were investigated from August 2000 to July 2002 in an oligotrophic lake, Lake Sapanca, in the northwest of Turkey. A total of 907 roach and 593 rudd individuals were examined. Macroscopic appearance of ovary, gonadosomatic index and oocyte diameter frequency distribution suggested that the spawning season of roach and rudd were from early April (13 °C) to late May (18 °C) and from early May (17.5 °C) to late June (23.5 °C), respectively. The spawning cycles of both species showed the same tendency in two successive years. The estimated mean total lengths at first maturity ( L T50) were 12.26 and 14.98 cm for male and female roach, respectively, and 7.12 and 8.10 cm for male and female rudd, respectively. The sizes at 50% maturity corresponded to 3 years for roach and 1 year for rudd, for both sexes. The ratio of males to females was 1.3:1 and 1.2:1 for roach and rudd, respectively. The average condition factor K was 1.21 for rudd and 1.14 for roach, and it was not affected by sex or reproductive effort. Absolute fecundity varied from 7829 to 95,387 eggs between ages 3 and 9 for roach and from 1807 to 35,629 eggs between ages 1 and 7 for rudd. Comparisons of these results with earlier studies from more northern regions revealed that spawning of both species in this area started earlier, but spawning temperature and the duration of spawning was approximately the same.  相似文献   

16.
Out of 19 Vibrio harveyi isolates obtained from a diversity of hosts and geographical locations, 14 were pathogenic to rainbow trout, Oncorhynchus mykiss (Walbaum), and Atlantic salmon, Salmo salar L., with mortalities of up to 100% following intraperitoneal injections of 106 cells fish?1. The extracellular products (ECPs) of only five pathogenic isolates were harmful to fish. Both pathogenic and non‐pathogenic cultures produced ECPs containing caseinase, gelatinase, phospholipase, lipase and haemolysins. Vibrio harveyi VIB 645, which was the most pathogenic isolate, produced ECPs with a maximal effect on salmonids from preparations obtained by using cellophane overlays on tryptone soya agar supplemented with 1% (w/v) sodium chloride with incubation at 28 °C for 24 h. This preparation contained the highest titre of haemolytic activity to Atlantic salmon (1:256) and rainbow trout (1:32) erythrocytes.  相似文献   

17.
This investigation examined the effects of temperature, density and early weaning on the survival and growth of Palaemonetes varians larvae. Survival of larvae raised at 17.5 °C was not significantly different (average + standard deviation) (94 ± 5%) from the survival of those raised at 19.5 °C (95 ± 5%) and at 21.5 °C (94 ± 4%). However, the duration of the larval stage was significantly longer for shrimp reared at 17.5 °C (17.3 ± 0.8 days) compared with shrimp reared at 19.5 °C (14.3 ± 0.7 days) and at 21.5 °C (11.3 ± 0.6 days). No significant differences ( P >0.05) were found in the survival rate, final weight and length of larvae reared at the densities of 5, 10, 20 and 50 larvae L−1. The survival of P. varians larvae fed solely on Artemia was significantly higher ( P <0.05) than larvae weaned with an artificial practical diet from Zoea II stage (94 ± 4% and 82 ± 1%, respectively, for Artemia and artificial diet-fed larvae), but no significant differences ( P >0.05) were observed in the final larval weight or length between these two treatments. The survival and growth of the larvae fed with the practical diet tested is a promising step ahead in the development of the culture of this species as it eliminates both the need for Artemia throughout all larval stages, and the need for more expensive artificial diets.  相似文献   

18.
ABSTRACT:   The optimal water temperature in seed germination and the upper critical water temperature in seedling growth were determined for Zostera marina collected from Ise Bay, Japan. The relationship between the seed germination rates and seed storage period (0, 30 and 60 days at 0°C) was also examined. The optimal water temperature for seed germination was in the range from 10 to 15°C regardless of the storage periods, in which germination rates ranged from 35 to 57%. Seedlings grown from seed up to 10 cm in total length were cultured for 1 week under various water temperatures to measure their relative growth rates. The optimal water temperature in growth was in the range from 20 to 25°C; relative growth rates ranged from 2.0 to 2.6%. Seedlings could survive up to a water temperature of 28°C, but most seedlings withered at 29 or 30°C. The optimal water temperatures for seed germination and seedling growth were related to the seasonal changes of water temperature at the sampling site. Although seedlings were rarely observed in the field in summer, they can grow at temperatures as high as 28°C. Therefore, Z. marina may extend its distribution as far as where the summer water temperature is lower than 28°C.  相似文献   

19.
ABSTRACT: Lethal limits of high temperature were studied to clarify the effects on the survival of the endangered Japanese crayfish species Cambaroides japonicus and the alien species Pacifastacus leniusculus . After the acclimation period for 2 weeks at 16°C, the temperature was raised at a rate of 1°C per week. As a result, the ultimate upper lethal temperatures of C.   japonicus and P.   leniusculus were 27.0 and 31.1°C, respectively, and the lethal temperature for P.   leniusculus was significantly higher than for C.   japonicus . The natural distributions of these two species are discussed in terms of the temperature tolerance.  相似文献   

20.
Ryohei  UENO  Naoto  URANO  Shigeru  KIMURA 《Fisheries Science》2002,68(3):571-578
ABSTRACT: A thermotolerant, fermentative yeast strain named RND13 from a hot spring drainage was evaluated for its ethanol-producing ability at elevated temperatures at a high substrate concentration [15% (w/v) glucose] close to the level reflecting industrial practice. The RND13 was capable of utilizing glucose almost completely at 40°C with increasing inoculum size, producing ethanol up to 6.6% (w/v), which is comparable to levels (7.0–7.2%) at 30°C. The maximum rate of ethanol production by the RND13 was found to be 9.0 g/L per h at 40°C in an inoculum sized 5% (w/v). At 43°C, however, the RND13 could not utilize glucose to completion and showed a slight drop in the extent of produced ethanol [6.0% (w/v)]. Thus, the culture at 40°C with a 5% cell inoculum was considered to be the optimal condition for ethanol production at higher temperatures in terms of batch fermentation. In the phylogenetic analysis based on the small-subunit rDNA sequence, the strain was grouped together with both Candida glabrata and Kluyveromyces delphensis , which are relatively close to Saccharomyces cerevisiae .  相似文献   

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