Étude de bactéries associées aux élevages de larves de bivalves marinsStudy of bacteria associated with the rearing of larvae of marine bivalves

Eighty-one bacterial strains associated with the rearing of marine bivalve larvae have been isolated from different origins: unicellular algal cultures; filtered sea water; unhealthy larvae; Mytilus edulis spawn, and contaminating strains from the laboratory atmosphere.The ecological, biochemical and nutritional characteristics and the sensitivity to antibiotics of these strains have been studied with the aid of 115 tests. This population appears to be similar to comparable populations studied by other authors.However, complementary studies taking into account the quantitative and qualitative variations of each source of bacterial contamination have been shown to be necessary, and are now in progress. When complete, more efficient bacterial control of the breeding of marine bivalve larvae will be possible.     

硅烷化表面海洋细菌对厚壳贻贝稚贝附着的影响

为探讨厚壳贻贝稚贝对自然微生物膜中海洋细菌的附着行为反应,本论文研究了厚壳贻贝（Mytilus coruscus）稚贝附着与硅烷基化附着基表面、微生物膜密度以及细菌种属系统发育之间的相互关系。研究表明所测海洋细菌均能显著促进厚壳贻贝稚贝的附着;其中Staphylococcus sp. 1和Cobetia sp. 1表现出较低诱导活性,且这两株细菌形成的微生物膜密度与稚贝附着率之间无显著相关性;其他7株海洋细菌均表现出中等程度诱导活性,且所形成的微生物膜密度与稚贝附着率之间呈显著相关性。系统发育分析表明所测海洋细菌对厚壳贻贝稚贝附着的诱导活性与细菌种属无关。因而,硅烷基化表面的海洋附着细菌对厚壳贻贝附着有着显著性促进作用,本研究将为后续开展厚壳贻贝稚贝附着机制奠定了基础。     

Relationships between intracellular bacteria and the bivalve killer dinoflagellate Heterocapsa circularisquama (Dinophyceae)

ABSTRACT: Relationships between the growths of five different clonal strains of Heterocapsa circularisquama and intracellular bacteria were investigated using culture experiments. Although each H. circularisquama strain culture was established from one cell by repeated and careful washings with micropipettes, intracellular and extracellular bacteria in each strain culture were still observed under a epifluorescence microscope using the DAPI (4',6-diamidino-2-phenylindole) staining technique. The extracellular bacteria are derived, presumably, from the inside of algal cells after the death and collapse of algal cells in culture. Using an electron microscope, bacteria were constantly observed in the cytoplasm and food vacuoles of H. circularisquama cells. The growth of five algal strains containing bacteria was compared with that of a bacteria-free strain using culture experiments under combined conditions of five different light intensities and five different strengths of culture medium. Bacteria showed no significant effect on the growth or survival of the algal cells. During the algal exponential growth phase, the intracellular bacterial cell numbers per algal cell decreased, whereas the total bacterial cell density in each algal culture increased. Final cell yield (total number) of the intracellular and extracellular bacteria varied considerably according to the algal strains. These results suggest that the intracellular bacteria of H. circularisquama grow or survive depending on the host alga, and that the alga can grow independently.     

Seasonal variations in biochemical composition and condition index of cultured mussels (Mytilus galloprovincialis Lmk) in the Lagoon of Venice (North Adriatic)

Seasonal changes in the main biochemical components and in the condition index of cultured mussels (Mytilus galloprovincialis Lmk) from the Venice Lagoon have been followed during the period October 1979–August 1981.Carbohydrates (2–32%) show winter minima and summer maxima, while proteins (30–52%) have a rather irregular pattern. Lipids (2–13%) and condition index exhibit an annual cycle with two minima (late spring; autumn-winter) and two maxima (early spring; summer).Seasonal variations of biochemical composition and condition index are discussed in connection with the reproductive cycle and some environmental parameters such as temperature and phytoplankton availability.The comparison between the present data and those from other geographic areas confirms the peculiarity of the environment of the Venice Lagoon.     

Humoral factors important in resistance of salmonid fish to bacterial disease. I. Serum antibody protection of rainbow trout (Salmo gairdneri) against vibriosis

Serum antibody is shown by passive immunization and absorption experiments to be an important component of defense mechanisms against experimentally induced vibriosis in rainbow trout (Salmo gairdneri). Antigen from heat-killed Vibrio anguillarum injected into trout with Freund's complete adjuvant induced much higher agglutinin titers than formalin-killed bacteria with or without adjuvant, or heat-killed bacteria without adjuvant. Passively transferred trout anti-Vibrio anguillarum serum provided protection against experimental vibriosis in trout for more than 2 months. Passively transferred anti-Vibrio anguillarum rabbit serum also protected trout.     

中湿度表面的海洋细菌对厚壳贻贝稚贝附着的影响

为研究自然微生物膜中海洋细菌对厚壳贻贝稚贝附着的影响,通过海洋贝类生物学、分子微生物生态学等手段调查附着基表面湿度、微生物膜的密度以及细菌种属系统发育与厚壳贻贝(Mytilus coruscus)稚贝附着关系。结果表明,所有测试海洋细菌形成微生物膜的最终密度随初始密度的增加而增加。所测海洋细菌均能不同程度地诱导厚壳贻贝稚贝的附着,其中Cobetia sp.3形成的微生物膜显示出最高诱导活性,其诱导的稚贝附着率为(70%±3%);Nautella sp.2、Pseudoalteromonas sp.9、Pseudoalteromonas sp.10、Bacillus sp.5和Pseudoalteromonas sp.11等5株细菌表现中等程度的诱导活性,其诱导的附着率范围为51%~60%。所有测试菌株所形成的微生物膜密度与稚贝附着均呈显著相关,尤其是Pseudoalteromonas sp.9和Cobetia sp.3诱导活性与附着率相关性极强,其相关性系数分别为0.774 1和0.723 3。系统发育分析结果表明,微生物膜密度和厚壳贻贝稚贝的附着率显著相关,然而海洋细菌诱导活性与细菌种属无关。因而,中湿度表面的海洋附着细菌对厚壳贻贝的附着有着明显的促进作用,本研究为后续开展厚壳贻贝稚贝的附着机制提供了理论依据。     

Distribution of three algicidal <Emphasis Type="Italic">Alteromonas</Emphasis> sp. strains in seagrass beds and surrounding areas in the Seto Inland Sea,Japan

Harmful algal blooms (HABs) often cause great damage to the fish aquaculture industry in the western part of Japan. Bacteria that effectively kill such phytoplankton are found in coastal seawater and are considered to influence the occurrence of HABs. Algicidal bacteria are found in abundance in seagrass beds; however, the distribution of particular bacterial strains has not yet been clarified. In this study, we determined the abundance of three algicidal Alteromonas sp. strains that were isolated as Chattonella antiqua-killing bacteria in seagrass beds and their surrounding areas in summers of 2013–2015. The strains were detected using a quantitative polymerase chain reaction (qPCR) amplification method. Two of the three algicidal bacterial strains were remarkably abundant in the seawater of an enclosed seagrass bed compared to the surrounding areas. In addition, the abundance of the three algicidal bacteria decreased when the eelgrass withered in late summer. These results suggest that growth of these algicidal bacteria was stimulated by eelgrass. The bacterial abundance estimated by qPCR was much greater than that determined by the culture-dependent method in June 2015 when the eelgrass grew thickly. This implies that the qPCR assay could be a sensitive tool to evaluate algicidal bacteria in natural environments.     

Elevage des larves de Loup (Dicentrarchus labrax, L.) a l'aide d'organismes du zooplancton congele: Resultats preliminairesRearing of sea-bass (Dicentrarchus labrax L.) larvae on frozen zooplankton: Preliminary results

Larvae (fry) of sea bass (Dicentrarchus labrax L.) were successfully reared from the mouth-opening stage to the age of 120 days being fed only on frozen zooplankton (Rotifera, Copopoda and Daphnia). This inert food is well accepted when left in the water less than 4 h after being thawed, but is rejected afterwards. The larval development can be compared to that obtained by artificial food and the survival rate is similar to that obtained by living plankton.     

Supplemental feeds containing coffee pulp for rearing Tilapia in Central AmericaAlimento suplementario conteniendo pulpa de café para la crianza de Tilapia en Centro America

A supplementary feed containing 30% coffee pulp was evaluated for use in the culture of Tilapia aurea in El Salvador, Central America. A comparison of the coffee pulp feed with a feed containing all of the same ingredients except coffee pulp was made with T. aurea raised in 1.0-m³ cages suspended in a fertilized earthen pond. Survival was high in all treatments and there was no significant difference in average weight gain between groups of fish receiving the two experimental feeds. Production trials were conducted in 100-m² fish pens and in 0.05-ha earthen ponds. Pen-raised fish receiving coffee pulp feed grew faster throughout the experiment, and total production was approximately twice that in control treatments. Highest production in pens was 1.25 kg/m² per year. Results of production pond trials using T. aurea at 9 000/ha and stocked with the piscivorous Cichlasoma managuense yielded an estimated 3 392 kg/ha per year in fed treatments and 2 049 kg/ha per year in controls. Low feed conversion (1.92) and low feed cost resulted in an increase in net annual earnings of $251.00/ha.     

Les antibiotiques dans les elevages de larves de bivalves marinsAntibiotics in larval rearing of marine bivalves

Fifty publications (1956–1976) concerning the use of antibiotics in rearing marine bivalve larvae are analysed. Most of the existing antibiotics have been tested alone or in combination, very frequently in an arbitrary fashion and without preliminary study to determine the most suitable antibiotic. The results obtained as regards to the survival of larvae or their growth have been variable, at times contradictory and not repeatable.The problems posed by the utilisation of antibiotics in rearing bivalve larvae are discussed and a standard routine for the use of antibiotics is proposed.     

Replacement of living microalgae with spray-dried diets for the marine mussel Mytilus galloprovincialis

Growth experiments were carried out in which juvenile mussels Mytilus galloprovincialis were fed diets composed of spray-dried algal products (Sanders Brine Shrimp) which included Schizochytrium sp. (SZ; Docosa Gold), Spirulina platensis (Sp) [=Arthrospira platensis] and Hematococcus pluvialis (H) either alone or in combination with partial rations of living microalgae. Growth of mussels fed 1/4 ration of living algae plus a mixed supplement of 50/50% w/w SZ:Sp grew significantly (SNK; P<0.05) faster than mussels fed a full live algal ration. Furthermore, growth (both wet weight and dry organic weight increase) of mussels fed diets composed of either 70/30% w/w SZ: ground Sp (GSp; <20 μm) or 25/50/25% w/w H:SZ:GSp was significantly (PLSD; P<0.05) greater than that of mussels fed an equal ration (by dry weight) of living Tahitian Isochrysis galbana.     

Probiotic bacteria <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> influences the blood profile in rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> (Walbaum)

This paper reports the effect of feeding probiotic diets on blood profiles in rainbow trout. Two experiments were performed: in the first, fish of average weight 75 g were offered either a commercial feed or the same incorporated with 10⁹ CFU g⁻¹ of lactic acid bacteria Lactobacillus rhamnosus for 30 days; in the second study performed for a similar duration, fish of average weight 126 g were offered formulated diets that either contained the same bacteria in heat-killed or freeze-dried form (nearly 10¹¹ CFU g⁻¹), or the basal diet without the bacteria. Blood samples were collected at different times after commencement of probiotic feeding to determine the total cholesterol, triglyceride contents, the plasma alkaline phosphatase activity, plasma protein and hematocrit value. The plasma cholesterol significantly increased upon probiotic feeding in the first experiment. A significant elevation (P < 0.05) of plasma cholesterol and triglyceride and alkaline phosphatase activity level was found in the freeze-dried probiotic fed groups at 20 and 30 days postfeeding. This was concomitant with the increased plasma protein and hematocrit values in FD group at 20 and 30 days. Likewise, the heat-killed probiotic fed group registered significantly high values of triglycerides, alkaline phosphatase activity, and plasma protein compared to the control diet fed groups after 20 days of feeding. Thus, alterations in the blood profiles could serve as supplementary information when examining the benefits of probiotics for fish.     

Etude de la croissance des juveniles de Chrysichthys walkeri (Günther) en etang en fonction de la densiteA study of the growth of juveniles of Chrysichthys walkeri (Günther) in ponds in relation to density

A study of the growth of Chrysichthys walkeri juveniles in relation to density was made with 2520 fry distributed in nine small square ponds. The study was successful in 5 ponds with 1490 individuals.The oxygen level of the water was low especially during the night. The temperature ranged between 28 and 30°C. The fry were stocked in ponds just after yolk vesicle resorption at a weight of 5.4 mg. After 4 weeks, the ponds stocked lightly showed better results (greater weight of the fishes) than the more densely stocked ponds. After 7 weeks, the growth difference between lots was very significant.The fishes were fed with pellets from the first day of the experiment. The total mortality was 38%.     

Use of a probiotic for the culture of larvae of the Pacific oyster (Crassostrea gigas Thunberg)

Additions of bacteria (strain CA2) as a food supplement to xenic larval cultures of the oyster Crassostrea gigas consistently enhanced growth of larvae during different seasons of the year. Bacterial enhancement of larval growth occurred when either Isochrysis galbana (ISO), I. aff. galbana (T-ISO) or Pseudoisochrysis paradoxa (VA-12) were used as algal foods. Additions of CA2 bacteria at 10⁵ cells ml⁻¹ to cultures of algal-fed larvae increased larval growth, the proportion of larvae that set to produce spat, and the subsequent size of spat. A lower proportion of slow-growing larvae in populations receiving additions of CA2 bacteria compared with populations of larvae fed only algae, suggests a bacterial nutritional contribution to larval growth. Manipulation of bacterial populations present in bivalve larval cultures is a potentially useful strategy for the enhancement of oyster production.     

Influence of Algal Exudates and Bacteria on Growth and Survival of White Shrimp Larvae Fed Entirely on Microencapsulated Diets

Intact microencapsulated protozoeal diets (MED), added to 5 m filtered hatchery water at the recommended concentration of 4 mg L^-1 gave no significant difference in bacterial numbers after 6 h, over a 48 h period compared to control groups in autoclaved hatchery water. Disruption of the MED resulted in a significant increase in bacterial numbers up to 12 h compared to controls with intact microcapsules. Protein leach rates were similar in sterile water and 5 m filtered water, indicating that leaching was independent of microbial activity. MED leached significantly less dietary solubles compared to a microparticulate diet (MPD). There was no evidence to suggest significant attachment of bacteria to diet particles for at least 48 h.In low-quality water after algal blooms, survival of white shrimp (Penaeus indicus) larvae was poor when reared in both sterile and 5 m filtered water. The addition of bacteria filtered from xenic algal cultures together with the algal exudates to such water gave significant improvements in survival to larvae fed MED in 5 m filtered water but not in autoclaved culture water. Live algal diets promote high larval survival and growth irrespective of the inclusion of bacteria.It is suggested that algal exudates act as selective bacteriocides and total replacement of algae with MED requires a balanced bacterial community which can be attained at times from natural sea water, or when the natural bacterial community balance is disturbed, corrected by the use of algal exudates.     

Evaluation of microalgae and industrial cheese whey as diets for Tapes decussatus (L.) seed: effects on water quality, growth, survival, condition and filtration rate

As reference diets for bivalve molluscs are missing, this study first evaluated the best mixed algal diet (Tetraselmis suecica Butch/Phaeodactylum tricornutum Bohlin) concentration for 4‐mm‐length Tapes decussatus (L.) clams. It also assessed the performance of an industrial by‐product (cheese whey) as a single diet or supplement for the best algal ration previously determined. Growth, survival, condition index and filtration rate were used to compare the different diets. Implications of rations tested on rearing water quality were also investigated, especially ammonium and heterotrophic bacteria levels. With a food concentration as algal dry weight per total seed live weight of 0.7% day^?1, seed for grow‐out (7 mm) could be obtained within 1 month. A mixture of 25% algal ration and 75% cheese whey gave the best performances. Artificial diets resulted in lower growth rates than live food (20 % to 40% of the best algal ration), but could be considered good maintenance rations in energetic terms. Using cheese whey, and without any algae, hatchery produced seed can be kept economically indoors for at least 30 days without loss in condition.     

添加外源益生菌对大菱鲆育苗生物饵料菌群结构的影响

为评价在大菱鲆育苗生产中添加外源益生菌对生物饵料轮虫和卤虫微生物菌群结构的影响,运用基于illumina HiSeq平台的高通量测序技术对添加益生菌和按照生产流程正常强化的轮虫、卤虫进行了菌群结构的分析和对比。添加外源益生菌的实验组轮虫和卤虫中的菌群物种多样性均明显高于对照组。在轮虫强化过程中,不同时期的对照组样品中菌群结构差异较明显,优势细菌种类变化较大。而添加外源益生菌后的各时期实验组轮虫菌群结构很相似,优势菌群的种类更为丰富,Lactococcus sp.、Pseudoalteromonas sp.和Alteromonas sp.等一直是各实验组中的优势细菌。在卤虫强化过程中,各对照组样品的菌群结构高度相似,优势细菌Cobetia sp.的相对丰度高达54%~65.2%。而在添加益生菌后,各实验组中的菌群结构仍高度相似,但Cobetia sp.的比例下降至4.3%~25.3%,最优势的细菌为Pseudoalteromonas sp.和Alteromonas sp.等几种,菌群结构中的物种均匀度更好。研究表明,在轮虫和卤虫强化过程中添加外源益生菌,能够改变生物饵料的菌群结构,使生物饵料中的细菌种类均匀度更好,并使菌群结构趋于稳定。     

Premiers resultats de l'elevage en cages de Tilapia nilotica (L.) dans le Lac Kossou,Cote d'IvoirePreliminary results of cage rearing Tilapia nilotica (L.) in Lake Kossou,Ivory Coast

The experimental rearing of T. nilotica in 1-m³ cages floating in Lake Kossou, Ivory Coast, was performed between March 1974 and April 1975. The fish were fed water-stable pellets (24.7% protein), the daily ration generally varying between 6 and 3.4% of the ichthyomass present. Fingerlings (9–55 g each) were stocked at densities of 200–400 fish/m³. They reached commercial size (more than 200 g) within 4–6 months, depending on the biomass present in the cage (stocking density and average individual weight of fish), as well as on limological conditions. The average annual mortality was 5.9%. Annual production varied from 36 to 64 kg/m³. The feed conversion rate averaged 2.8. A reduction of the daily ration for fish larger than 40 g reduced this value.Fingerlings should average at least 20–30 g in weight at initial stocking of the cage. Two crops a year can then be easily realized. If only male fish are reared, three harvests per year appear to be possible, increasing the annual cage production of fish close to 200 kg/m³ with proper management.The possible development of T. nilotica cage culture is actually hampered due to the lack of adequate fish feed available commercially, and to the lack of sufficient quantities of calibrated fingerlings throughout the year.     

Use of a <Emphasis Type="Italic">Noctiluca</Emphasis>-killing bacterium <Emphasis Type="Italic">Marinobacter salsuginis</Emphasis> strain BS2 to reduce shrimp mortality caused by <Emphasis Type="Italic">Noctiluca scintillans</Emphasis>

The ability of an algicidal bacterium Marinobacter salsuginis strain BS2, isolated from shrimp pond water, to reduce shrimp mortality was investigated under laboratory conditions. When two species of shrimp (Penaeus monodon and Litopenaeus vannamei) (body length 1.5–1.8 cm) were cultured together with the dinoflagellate Noctiluca scintillans, nearly 80 % of the shrimps died within 7 days. However, when bacterial strain BS2 was also added to the culture, N. scintillans was killed within 48 h, and shrimp survival rates on the 7th day improved from 23 to 87 % for both P. monodon and L. vannamei. The bacterium BS2 alone had no effect on shrimp condition. Under conditions of increased dissolved oxygen, the effect of using BS2 was greater, and shrimp survival rates improved even more dramatically, from 26 to 98 %. These studies provide the first evidence that the use of killing bacteria, isolated from shrimp culture water, can suppress harmful algal blooms (HABs) and thus restore the efficiency of shrimp production. The control of HABs in this way in shrimp culture farms would be a major benefit for shrimp production.     

Characterisation of the microbiota associated with intestine of Atlantic cod (Gadus morhua L.): The effect of fish meal, standard soybean meal and a bioprocessed soybean meal

Populations of aerobic heterotrophic bacteria present in the gastrointestinal (GI) tract of healthy Atlantic cod (Gadus morhua L.) fed three different diets, fish meal, standard or a bioprocessed soybean meal (BPSBM), were estimated using the dilution plate technique. A total of 944 isolates were characterised by biochemical and physiological properties and 425 isolates were identified further by sequencing the 16S rRNA genes. Our results showed that gut microbiota were affected by dietary manipulation. The GI tract of fish fed fish meal was dominated by Gram-positive bacteria of the genera Brochothrix and Carnobacterium. The Gram-negative bacteria Chryseobacterium spp. and Psychrobacter glacincola, and Gram-positive bacteria belonging to Carnobacterium, dominated in the digestive tract of fish fed soybean meal. In contrast to these results, genus Psychrobacter dominated in the GI tract when fish were fed BPSBM.Until recently, it was generally suggested that the gut microbiota of fish were less diverse than in homoeothermic animals. However, the present study identified several “new” bacterial species isolated from the alimentary tract of Atlantic cod. These “new” bacterial species are not normally isolated from the GI tract of fish. Based on our finding we suggest that the GI tract microbiota of fish might not be as simple as believed.Antagonistic activity of carnobacteria regarding inhibition of growth of two fish pathogens (Aeromonas salmonicida ssp. salmonicida and Vibrio anguillarum) was observed. However, some difference in the antibacterial activity of Carnobacterium spp. was observed. Whether this antagonistic activity has any effect in challenge studies will be discussed, especially in relation to the finding that the digestive tract is one of the major infection routes in fish.