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1.
As adhesion and translocation through fish gut enterocytes of the pathogen Vibrio (Listonella) anguillarum are not well investigated, the effective cause of disease and mortality outbreaks in larval sea bass, Dicentrarchus labrax, suffering from vibriosis is unknown. We detected V. anguillarum within the gut of experimentally infected gnotobiotic sea bass larvae using transmission electron microscopy and immunogold labelling. Intact bacteria were observed in close contact with the apical brush border in the gut lumen. Enterocytes contained lysosomes positive for protein A‐gold particles suggesting intracellular elimination of bacterial fragments. Shed intestinal cells were regularly visualized in the gut lumen in late stages of exposure. Some of the luminal cells showed invagination and putative engulfment of bacterial structures by pseudopod‐like formations. The engulfed structures were positive for protein A‐colloidal gold indicating that these structures were V. anguillarum. Immunogold positive thread‐like structures secreted by V. anguillarum suggested the presence of outer membrane vesicles (MVs) hypothesizing that MVs are potent transporters of active virulence factors to sea bass gut cells suggestive for a substantial role in biofilm formation and pathogenesis. We put forward the hypothesis that MVs are important in the pathogenesis of V. anguillarum in sea bass larvae.  相似文献   

2.
Larval mortalities occurring in molluscan hatcheries have often been associated with bacterial contamination, and more specifically with vibrios. Although batches of oyster and clam larvae have been routinely reared in the hatchery of Argenton (North Brittany, France) without antibiotics, high larval mortalities have been recorded with the great scallop, Pecten maximus, under similar conditions. For this species, an addition of antibiotics was found necessary and chloramphenicol was used at a concentration of 8 mg l–1. However, this chemical has now been banned in Europe, making either substitution products or an improvement in the rearing procedures essential. Studies carried out have shown that neither a decrease in larval density (to 1 larva ml–1) nor an increase in seawater change frequency (to one per day) had any positive effects. Furthermore, elective substances such as sugars were not suitable and the use of another antibiotic, erythromycin, led to inconsistent results. The only positive effects were obtained with lower levels of chloramphenicol, which does not resolve the problem. Because no alternative solutions have as yet been found, further research needs to be undertaken.  相似文献   

3.
During a series of experiments, bacteriological elements in scallop larval rearing were investigated: larvae susceptibility to pathogens as a function of their age, and the use of probiotic bacteria during larviculture. Younger larvae (d5 PF) were highly more susceptible to pathogenic‐challenge than their older siblings, which were challenged at an older age (d15 PF). A challenge with 104 CFU mL?1 of V. pectenicida killed 100% of d5 PF larvae 7 days following challenge, yet killed only 9% of d15 PF larvae 9 days following challenge. Use of the probiotics Phaeobacter gallaeciensis, Alteromonas macleodii 0444 and Neptunomonas sp. 0536, provided for larger larvae, a high yield of competent larvae and, perhaps more importantly, protection against pathogen‐challenge similar to levels achieved from antibiotic use. When challenged with V. pectenicida, d29 survivals were 20.3%, 85.1% and 75.0% respectively for control (no probiotic), antibiotic treated, and ‘probiotic mix’ administered larvae. Use of potential probiotic Pseudoalteromonas sp. D41 appeared to hinder scallop larvae. Future use of probiotics in scallop larval rearing would benefit from combined use of P. gallaeciensis, A. macleodii 0444 and Neptunomonas sp. 0536.  相似文献   

4.
The development of sustainable methods for sea urchin juvenile production is currently constrained by high mortality rates during larval growth and the high costs of larval rearing systems management. With the aim of developing a method for the production of juveniles of the purple sea urchin Paracentrotus lividus in a medium‐scale recirculating system, the present study focused on the effects of high stocking densities on larval growth. Plutei larvae were reared at three different densities (up to 7 ind/ml) in a semi‐static culture system. Larval survival and metamorphosis success were evaluated in order to identify the most effective density range. The highest metamorphosis rates (80%–95%) were obtained at 4 and 7 larvae/ml. These results are comparable with (and in some cases higher than) those reported for the same species at much lower larval densities. In conclusion, the rearing conditions tested here show for the first time that a significantly higher (4 ind/ml) stocking density than those of traditional P. lividus rearing methods (based on large volumes and low densities) can be adopted, thus supporting the feasibility of an increase in the final output of competent larvae with no increase in rearing volumes.  相似文献   

5.
Surveys for bacteriological analysis of larval samples to isolate the associated vibrios were carried out during 1985–1992, 2001 and 2002 in three different hatcheries located on the southwest coast of India. Vibrio isolates were examined for their species diversity, virulence based on haemolysis in prawn blood agar, lipolysis, proteolysis and chitinolysis and antibiotic sensitivity. Vibrio cholerae was the predominant species in the apparently healthy larval samples, whereas V. alginolyticus and V. vulnificus dominated during disease and morbidity. No correlation was found between the hydrolytic properties and haemolytic activity of the vibrios associated with the larvae. All isolates were resistant to erythromycin and resistance to oxytetracycline, ampicillin and streptomycin sulphate was prevalent among the larger section of the Vibrio population. This suggested that antibiotic application may not be of much use to protect the larvae from vibriosis. This is the first report on the diversity of Vibrio species associated with Macrobrachium rosenbergii larvae and their virulence characteristics based on haemolysis in prawn blood agar.  相似文献   

6.
Controlled incorporation of selected microalgae and bacteria in aquaculture systems can be beneficial because they can act as microbiological control. That is why the characteristics of biofilm generated naturally in Seriola lalandi culture cages were analysed, their potential benefit to the growth of larvae was studied, and their controlled use for improving the larval viability and as a vector to improve incorporation of previously studied probiotic bacteria was tested. According to biodiversity results, these biofilms are composed of a diatom–bacteria mix showing a decrease in biodiversity in laboratory culture conditions being dominated by Navicula phyllepta and bacteria of the family Rhodobacteraceae. This can be produced on mesh substrates incorporated in bioreactors with rapid growth rate and adhesiveness. Preliminary results from the addition of substrates with this specific biofilm in larvae culture systems showed that it is consumed by the larvae without negative effects, while positive effects on the viability of larvae in combination with probiotics were observed. Considering preliminary results, the addition of these specific substrates with diatom–bacteria biofilms could be a good improvement for aquaculture systems and together with the use of probiotics can contribute to maintaining a stable and controlled system improving the viability of the larval fish culture in its early stages.  相似文献   

7.
Veliger larvae of the mussel Mytilus edulis from the Menai Strait, North Wales, Great Britain, were reared for 15 days in the presence of copper and plastic strips painted with one of four paint types. Severe larval mortalities occurred when the paints had only been allowed to leach in sea water for 3 days prior to the rearing trials. However, when the paints had been leached for 30 days, there were no significant effects on growth or mortality of the larvae. None of the paints allowed copper ions to permeate into the sea water of the larval cultures.  相似文献   

8.
A bacteriological study was carried out at a scallop (Pecten maximus) hatchery near Bergen, western Norway following a severe increase in mortality rates during the larval stages of the scallops. No larvae survived to settling, except for those in groups treated prophylactically with chloramphenicol. In order to identify pathogenic strains of bacteria, we performed a challenge test on 10- to 16-day-old larvae using isolated bacterial strains from the hatchery. Infection with six of these strains produced mortalities that were not statistically different from that resulting from infection with the known pathogen Vibrio pectenicida. However, about 5% of the strains tested in the challenge experiment produced higher motility rates than found in the unchallenged control group, indicating a possible probiotic effect. On the basis of 16S rDNA analysis on these strains, the phylogenetic tree indicated two groups of apparent pathogens: (1) one strain, LT13, grouped together with Alteromonas/Pseudoalteromonas; (2) a cluster of strains grouped together with Vibrio splendidus (LT06, LT21, LT73, PMV18 and PMV19). Strain LT13 was isolated from cultures of the microalga Chaetoceros calcitrans used for feed, while the other strains were isolated from larval cultures. Transmission electron microscopy showed intracellular bacteria that resembled bacteria in the groups Chlamydiaceae and Rickettsiaceae.  相似文献   

9.
Egg capsules of Concholepas concholepas were analysed microbiologically, showing the presence of bacteria inside the capsules. The bacteria were isolated and identified, and their probiotic potential on early larvae of C. concholepas was evaluated. From 32 capsules that were examined, 45 bacterial strains were isolated, of which 67% belonged to the genus Bacillus. Survival bioassays were carried out with all the isolated bacterial strains on C. concholepas larvae. It was found that 71% of the bacteria had positive effects on larval survival. This research determined that bacteria of the genera Bacillus and Vibrio present in the capsules have probiotic potential. The bacteria that led to the greatest larval survival were Bacillus pumilus, which may be used in the cultivation of C. concholepas, because one of the limiting factors for the cultivation of this species is larval survival; thus, we propose that the probiotic strains identified in this study will be used during the cultivation of C. concholepas to enhance the chances of larval survival.  相似文献   

10.
隋炜金  王鸿霞  刘保忠 《水产学报》2023,47(6):069413-069413
为研究文蛤育苗过程中幼虫病害及其主要致病菌,实验通过构建细菌16S rDNA克隆文库、病原菌分离纯化、人工感染和药敏实验等方法对育苗场发病的浮游期文蛤壳顶幼虫样品进行了系统分析。结果显示,发病的文蛤壳顶幼虫细菌群落多样性低,地中海弧菌占比高达75%以上,推断其可能为引发此次幼虫发病的主要致病菌。从发病幼虫的匀浆组织中分离获得该优势菌株,测序及系统发育鉴定为地中海弧菌。人工感染实验确定了其致病性,菌株US2-01在1.0×106 CFU/mL的菌液浓度下浸泡感染文蛤浮游幼虫,96 h累计死亡率为84%。药敏实验表明,地中海弧菌菌株US2-01在12种抗生素的测试中对常用的青霉素、氨苄西林、红霉素等抗生素具有一定的耐药性,对四环素和多西环素中度敏感,对头孢他啶、庆大霉素、卡那霉素等其余7种抗生素呈现高度敏感。本研究首次报道了地中海弧菌为文蛤浮游期幼虫致病的一种潜在病原,研究结果可为文蛤幼虫疾病研究及贝类苗种培育过程中的病害防控提供科学依据。  相似文献   

11.
A major concern in larvae production is a mass mortality caused by fish diseases. In larvae production, pumped‐up natural seawater filtered through a sand filter system is used for fish rearing, and microalgae and rotifer cultures. Here, we investigated the community structures of eukaryotic microbes, as well as total bacteria and vibrios, in various processed ‘waters’ used in a larvae production site. We observed that ultraviolet irradiation of seawater was effective to reduce not only total bacteria and vibrios but also eukaryotic microbes. Moreover, the community structures of total bacteria and vibrios in rearing waters for fish larvae were different from those in rotifer cultures fed with Chlorella, but rather similar to those in natural seawater and microalgae cultures. These results suggest that the bacterial community in rearing waters may originate mainly from natural seawater and then be selected by microalgae in rearing water. Overall, this study provides useful information for avoiding the risk of fish disease outbreaks in a larvae production site.  相似文献   

12.
Vibrionaceae infections are a major obstacle for marine larviculture; however, little is known about virulence differences of Vibrio strains. The virulence of Vibrio strains, mostly isolated from vibriosis outbreaks in farmed fish, was tested in larval challenge trials with cod (Gadus morhua), turbot (Scophthalmus maximus) and halibut (Hippoglossus hippoglossus) using a multiwell dish assays with single‐egg/larvae cultures. The strains differed significantly in virulence as some caused a high mortality of larva reaching 100% mortality after a few days, while others had no or only marginal effects on survival. Some Vibrio strains were pathogenic in all of the larva species, while some caused disease only in one of the species. Twenty‐nine of the Vibrio anguillarum strains increased the mortality of larvae from at least one fish species; however, pathogenicity of the strains differed markedly. Other Vibrio species had no or less pronounced effects on larval mortalities. Iron uptake has been related to V. anguillarum virulence; however, the presence or absence of the plasmid pJM1 encoding anguibactin did not correlate with virulence. The genomes of V. anguillarum were compared (D. Castillo, P.W. D'Alvise, M. Middelboe & L. Gram, unpublished data) and most of the high‐virulent strains had acquired virulence genes from other pathogenic Vibrio.  相似文献   

13.
An alternative larval shrimp feeding regime, in which umbrella‐stage Artemia were constituting the first zooplankton source was evaluated in the culture of Litopenaeus vannamei. In a preliminary experiment, umbrella‐stage Artemia were fed to larvae from zoea 2 (Z2) to mysis 2 (M2) stages to identify the larval stage at which raptorial feeding starts and to determine daily feeding rates. The following experiment evaluated the performance of two feeding regimen that differed during the late zoea/early mysis stages: a control treatment with frozen Artemia nauplii (FAN), and a treatment with frozen umbrella‐stage Artemia (FUA). The ingestion rate of umbrella‐stage Artemia increased from nine umbrella per larvae day?1 at Z2 stage to 21 umbrella per larvae day?1 at M2. A steep increase in ingestion and dry weight from Z3 to M2 suggests a shift to a raptorial feeding mode at the M1 stage. Treatment FUA exhibited a significantly higher larval stage index (P < 0.05) during the period that zoea larvae metamorphosed to the mysis stage, and a higher final biomass, compared with treatment FAN. Based on these results and on practical considerations, a feeding regime starting with umbrella‐stage Artemia from Z2 sub‐stage can be recommended for L. vannamei larvae rearing.  相似文献   

14.
Larval growth, age, growth effect and instantaneous mortality were estimated in anchoveta, Engraulis ringens, collected biweekly during the austral winter of 2014 in nearshore waters off Bay of Antofagasta (23°41′W–70°30′S), northern Chile. Through measuring standard length (SL) and sagitta microstructure analysis, it was estimated that the growth rate of E. ringens larvae decreased from June (0.85 mm day?1) to August (0.50 mm day?1). However, the water temperature was homogeneous during the sampling dates (14.6, 15.2, 14.4, and 14.6°C), suggesting that the decelerating larval growth was not linked to changes in sea temperature. Additionally, larvae with slow growth have larger otoliths compared with conspecifics with fast growth (growth effect). Larval mortality rates tended to decrease until the middle of July (0.18 per day) but increased to 0.25 per day in early August, which coincided with lower food availability (i.e., chlorophyll‐a, 2.7–5.6 mg m?3) and a high occurrence of smaller larvae (1.58–11.5 mm). Partial least squares analysis indicates low covariance between the biological and oceanographic variables (PLS: 11.71%), suggesting that other factors, such as parental effects, may explain the abrupt decrease in the larval growth rates.  相似文献   

15.
An optimal chemical shell marking protocol was developed for the New Zealand green‐lipped mussel, Perna canaliculus with a view to its future use in larval tracking experiments. Larval P. canaliculus aged either 10, 15 or 19 days post fertilization were immersed in treatments of 50, 100 and 200 mg L?1 of calcein for a period of 24 h before measurements of shell mark brightness were taken. There was 100% marking success in all calcein treatments for all age classes, with 19‐day larvae immersed in 200 mg L?1 calcein producing the brightest mark. Growth was not affected by calcein immersion; however, 10‐day larvae exhibited significantly higher levels of mortality compared with 15‐ and 19‐day larvae suggesting a reduced resilience to the marking protocols in younger larvae. In a mass staining experiment, a solution of 100 mg L?1 calcein was used to successfully stain15.6 million hatchery reared P. canaliculus larvae. Calcein, therefore, offers a low impact method with which to stain the sensitive early life stages of this species thus providing a rapid method for identifying individuals of interest, i.e. individuals released in the wild or specific family lines within a hatchery environment.  相似文献   

16.
《水生生物资源》1999,12(1):37-44
Both in Indonesia and Vietnam, larval rearing of Pangasius hypophthalmus remains problematic due to the variable and generally low survival rates obtained. The first week represents the most critical period for these larvae and up to now, cannibalistic behaviour was considered as the main cause of mortality. In the present study, two experiments were carried out in order to better understand the evolution and causes of mortality of P. hypophthalmus larvae from hatching up to 8 d of age and to find measures to improve survival rates. The first experiment was designed to evaluate the importance of cannibalism and differences in mortality when larvae from two different females were reared either in groups of 30 individuals or in isolated condition (30 larvae reared separately). In both cases, the culture was carried out either with or without antibiotic. The aim of the second experiment was to test oxytetracycline and different disinfectants (chloramine-T, formalin and ‘formalin + malachite green oxalate’) at different dosages in order to prevent bacterial outbreaks in the culture. The results indicated that the survival rates of P. hypophthalmus larvae was dependent on the initial quality of larvae or eggs and that larval mortality was more a consequence of pathogenic infection than a direct effect of cannibalism. The present study demonstrated that the survival rates of larvae were systematically improved when rearing was carried out in water containing antibiotic (oxytetracycline at a dose of 5 to 20 mg·L1). Survival rate and final mean body weight of larvae as high as those obtained using antibiotic were also reached with applications of disinfectants such as chloramine-T and formalin. The use of these disinfectants is recommended for commercial P. hypophthalmus hatcheries.  相似文献   

17.
Amphidromous fish species reproduce in rivers, and their larvae immediately drift to the sea after hatching. Most of these larvae survive in freshwater for a few days only, rapidly reaching sea water is thus essential. Being of small size, especially among species of the Sicydiinae subfamily, the larvae possess poor swimming abilities; their drift dynamics is considered to be mainly passive and influenced by flow conditions. The influence of daily fluctuations in flow regime on Sicyopterus lagocephalus (Sicydiinae) larval drift patterns was studied using drift nets at three sites along the dammed Langevin River in Reunion Island. The river flow was stable at the upstream site when frequent anthropogenic river flow fluctuations were observed at the two sites downstream to the dam. Weak diel larval drift dynamics were detected at the upstream site and the first site under anthropogenic river flow conditions. In contrast, larval drift dynamics at the most downstream site was strongly influenced by anthropogenic daily fluctuations of discharge: the abundance of drifting larvae increased with peaks of discharge, regardless of the time. As higher mortality rates of drifting larvae were observed during discharge peaks, the benefit of a more rapid travel to the ocean associated with these peaks should be lower than expected. It is concluded that peaks of discharge increase the number of larvae drifting to the sea probably due to higher physical constraints on the egg clutches, but also decrease the chance of survival for larvae that may be nonfully developed.  相似文献   

18.
The European clam, Ruditapes decussatus is a species with high commercial importance in Portugal and other Southern European countries. However, the development of R. decussatus culture has been limited by the highly variable patterns of natural recruitment. The development of hatchery technology will provide an alternative source of spat. The effect of six nutritional regimes on the survival, growth and biochemical composition of R. decussatus larvae were evaluated, aiming to provide crucial information on its nutritional requirements. A holistic approach incorporating all physiological response showed that the bispecific diet I. aff galbana and C. calcitrans (60/40 cell μL?1) was the more adapted throughout larval development. Moreover, the monospecific diet I. aff galbana provided an overall good performance. Larvae cannot use C. calcitrans at early stages of development; however, the inclusion of these microalgae improved late larval development. The results obtained constitute an important first step in the hatchery R. decussatus larval nutrition and a prerequisite for future work on the improvement of larval development and the optimization of feeding practices that will maximize larvae yield and minimize cost in aquaculture hatcheries.  相似文献   

19.
Hatchery culture of the razor clam, Solen marginatus (Pulteney), has recently been developed in Galicia (NW Spain). However, recurrent episodes of mortalities of larval and post‐larval cultures have been recorded during the course of various studies. The disease signs were similar to those described for other bivalve species in outbreaks caused by bacteria of the genus Vibrio. In this article, we present the results of microbiological monitoring of two batches of razor clams with different survival rates. All fermentative isolates were identified as members of the Splendidus clade within the genus Vibrio. Some of these isolates, identified as Vibrio splendidus‐like, were clearly associated with the batch suffering mortalities, indicating their possible role as pathogens. Similar strains were found in the broodstock, suggesting vertical transmission of these bacteria. This is the first study of the microbiota associated with hatchery culture of S. marginatus, and the results will provide useful information for the optimization of a protocol for hatchery culture of this bivalve species.  相似文献   

20.
The quality of the microalgae provided on Paracentrotus lividus larvae rearing is a primordial factor having a direct (nutritional properties) and indirect (water quality) impact on growth, competence and survival. Skeletonema costatum is a diatom commonly used in the bivalve cultivation. However, the use of this diatom in P. lividus larval cultivations is poorly known. The Rhodomonas spp. is a microalgae commonly used in sea urchin larvae culture. Three different diets were tested on P. lividus larvae and post‐larvae cultivation (D1—Rhodomonas marina, D2—S. costatum, D3—mixture of both algae). Larvae fed with the D2 diet (55.8%) and D3 (39.9%) had a survival at 15 DAH higher than D1 (5.5%). The low survival in D1 could be due to the higher microbiological load on microalgae (Vibrio alginolyticus and V. pectenicide). Larvae fed with S. costatum (D2) showed a lower development than other diets. The competency index was lower for larvae fed with the D2. These results show that microalgae diversified diets contribute to a better development of P. lividus larvae. During the settlement and post‐settlement phase, there was also a lower growth of the sea urchin fed with the D2 and a higher survival for D3.  相似文献   

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