几株益生菌对鲤前肠黏液的体外黏附特性

采用体外固定鲤前肠黏液,结合细菌同位素示踪的方法,对来源于鲤肠道的2株肠球菌和1株柠檬酸杆菌以及养殖水体的2株芽孢杆菌的细菌表面凝集素和黏液蛋白受体的化学组成、对病原菌附着鲤前肠黏液的影响等进行了研究。结果表明,5株菌经高碘酸钠和蛋白水解酶修饰后,高碘酸钠能极显著降低芽孢杆菌、肠球菌和柠檬酸杆菌的黏附率(P<0.01),而蛋白水解酶对多数菌株的影响不显著(P>0.05),推测细菌表面的凝集素主要为具糖蛋白性质的物质。黏液蛋白经蛋白水解酶处理后,部分菌株的附着数量显著降低(P<0.05),但经高碘酸钠处理后,5株菌的黏附率均显著上升(P<0.05),提示黏液蛋白上存在的5株细菌的特异性受体物质可能为蛋白质。益生菌通过排斥、竞争和取代作用能够显著降低部分病原菌的黏附率。取代作用对病原菌黏附的抑制效果最好,但具有菌株特异性。     

Effect of dietary administration of probiotics on growth and intestine functionality of juvenile Senegalese sole (Solea senegalensis, Kaup 1858)

The effects of the dietary administration of two bacterial probiotic strains (Ppd11 and Pdp13) from the Alteromonadaceae family for 60 days, were assessed by measuring growth and feed efficiency, activities of leucine aminopeptidase and alkaline phosphatase and structural changes in the intestine of juvenile Senegalese sole. In addition, the profile of intestinal microbiota was studied by Denaturing Gradient Gel Electrophoresis. Growth and nutrient utilization were significantly higher in fish receiving probiotics than in those fed the control diet. No differences were observed in proximal composition between treatments, though higher lipid muscle content was measured in fish receiving Pdp13. Those fish also exhibited higher activities of AP when compared to Ppd11 and control groups. The profile of intestinal microbiota clearly separated those fish receiving probiotics from those of the control group. Microscopical examination revealed accumulation of lipid droplets in the enterocytes of fish receiving the control diet, but not in those fed on probiotics. Interactions between those structural changes and growth performance are discussed.     

Screening of gastrointestinal bacteria of Indian major carps for a candidate probiotic species for aquaculture practices

Probotic selection could be detrimental for the success of aquaculture practices. While, in most of the cases emphasis is given to the production of antibacterial compounds, and in vitro inhibitory activities of a bacterium for choosing a probiotic strain, however, other criteria like non‐antibiotic resistant, non‐haemolytic nature and non‐pathogenic potentiality of probiotic strain are often overlooked. We have screened gastrointestinal bacteria isolated from all the three species of Indian major carps viz., Labeo rohita, Catla catla and Cirrhinus mrigala for a suitable probiotic. Bacteria like Aeromonas (Aeromonas species, A. hydrophila), Micrococcus, Corynebacterium, Plesiomonas, Bacillus (Bacillus species, B. subtilis) and Pseudomonas (Pseudomonas species, P. aeruginosa) exhibited antibacterial activities against different pathogens. Among these, B. subtilis was found to grow at a wider range of temperatures, pH and salt concentrations and was non‐haemolytic, non‐antibiotic resistant, non‐pathogenic and inhibited all the tested primary and secondary pathogens. Dietary supplementation of the bacteria at 10⁸ CFU g^?1 feed also enhanced the growth of Indian major carp, L. rohita. Based upon these characteristics, we aimed to develop a suitable probiotic for aquaculture practices.     

Dietary microorganism and plant effects on the survival and immune response of Litopenaeus vannamei challenged with the white spot syndrome virus

The effect of plants and probiotics on the survival and immune response of Litopenaeus vannamei challenged with the white spot syndrome virus (WSSV) was evaluated. A probiotic mixture (PM), plant extract (PE) or powdered plants (PP) were added to feed with the attractant Dry Oil^®. An experiment was conducted with five treatments in triplicate. Shrimp (weighing 11.70±2.5 g) were cultured in 120 L plastic tanks and fed twice a day with commercial feed plus additives or with commercial feed plus WSSV. Animals were monitored for the occurrence of WSSV using single‐step and nested PCR. The PM and PP added to the commercial feed showed high survival, a decrease in WSSV prevalence in shrimp and an increase in the activity of lysosomal enzymes, N‐acetyl‐β‐glucosaminidase and acid phosphatase. The total haemocyte count in shrimp treated with PM was significantly higher than that in the control group (treatment I) and in shrimp fed with PE. The results of the present work indicate that PP and PM are good candidates for use as feed additives against WSSV in shrimp cultures.     

Carnobacterium maltaromaticum vs. Vibrio (Listonella) anguillarum in the midgut of Atlantic cod (Gadus morhua L.): an ex vivo study

The present investigation addresses whether the midgut (MG) of Atlantic cod (Gadus morhua L.) is an infection route for Vibrio (Listonella) anguillarum serotype 02 β and if Carnobacterium maltaromaticum, a probiotic bacterium, can out‐compete the pathogen and modulate the autochthonous MG microbiota. This was investigated by using an ex vivo method the intestinal sac, utilized previously in studies on Atlantic cod and Atlantic salmon (Salmo salar L.). Exposure of the MG to V. (L.) anguillarum did not reveal any cell damage indicating that the MG does not appear to be an infection route for V. (L.) anguillarum in healthy Atlantic cod. This finding together with previous observations on Atlantic cod and Atlantic salmon indicate that intestine as an infection route might vary between these two species. When the MG was exposed to C. maltaromaticum, no cell damage or cellular disruptions were observed. As budding from the apices of microvilli was observed in all treatments exposed to bacteria, we suggest that budding might be involved in the primary barrier against bacterial infection. However, to clarify this hypothesis, further studies are needed. Exposure of the MG to the probiotics and pathogenic bacteria indicated that C. maltaromaticum, to some extent, is able to out‐compete V. (L.) anguillarum but the topic merits further investigation. Analysis of the MG microbiota after sterile saline solution and bacterial exposure indicates that bacteria related to Staphylococcus sciuri belong to the autochthonous gut microbiota in Atlantic cod.     

Lactic acid bacteria vs. pathogens in the gastrointestinal tract of fish: a review

Intensive fish production worldwide has increased the risk of infectious diseases. However, before any infection can be established, pathogens must penetrate the primary barrier. In fish, the three major routes of infection are the skin, gills and gastrointestinal (GI) tract. The GI tract is essentially a muscular tube lined by a mucous membrane of columnar epithelial cells that exhibit a regional variation in structure and function. In the last two decades, our understanding of the endocytosis and translocation of bacteria across this mucosa, and the sorts of cell damage caused by pathogenic bacteria, has increased. Electron microscopy has made a valuable contribution to this knowledge. In the fish‐farming industry, severe economic losses are caused by furunculosis (agent, Aeromonas salmonicida spp. salmonicida) and vibriosis [agent, Vibrio (Listonella) anguillarum]. This article provides an overview of the GI tract of fish from an electron microscopical perspective focusing on cellular damage (specific attack on tight junctions and desmosomes) caused by pathogenic bacteria, and interactions between the ‘good’ intestinal bacteria [e.g. lactic acid bacteria (LAB)] and pathogens. Using different in vitro methods, several studies have demonstrated that co‐incubation of Atlantic salmon (Salmo salar L.) foregut (proximal intestine) with LAB and pathogens can have beneficial effects, the cell damage caused by the pathogens being prevented, to some extent, by the LAB. However, there is uncertainty over whether or not similar effects are observed in other species such as Atlantic cod (Gadus morhua L.). When discussing cellular damage in the GI tract of fish caused by pathogenic bacteria, several important questions arise including: (1) Do different pathogenic bacteria use different mechanisms to infect the gut? (2) Does the gradual development of the GI tract from larva to adult affect infection? (3) Are there different infection patterns between different fish species? The present article addresses these and other questions.     

Probiotic effect of Bacillus NL110 and Vibrio NE17 on the survival,growth performance and immune response of Macrobrachium rosenbergii (de Man)

Eight hundred and eighty‐five strains of bacterial isolates from various samples associated with the natural habitat of Macrobrachium rosenbergii were screened for their probiotic potential. Two putative probionts namely Bacillus NL110 and Vibrio NE17 isolated from the larvae and egg samples, respectively, were selected for experimental studies and were introduced to the juveniles of M. rosenbergii (0.080±0.001 g) through different modes such as through feed, water and both. The probiotic potential of the above bacteria in terms of improvements in water quality, growth, survival, specific growth rate (SGR), feed conversion ratio and immune parameters was evaluated. The treatment groups showed a significant improvement in SGR and weight gain (P<0.001). Survival among different treatment groups was better than that in the control group. There were also significant improvements in the water quality parameters such as the concentration of nitrate and ammonia in the treatment groups (P<0.05). Improvements in immune parameters such as the total haemocyte count (P<0.05), phenoloxidase activity and respiratory burst were also significant (P<0.001). It is concluded that screening of the natural microflora of cultured fish and shellfish for putative probionts might yield probiotic strains of bacteria that could be utilized for an environment‐friendly and organic mode of aquaculture.     

Effects of probiotic Pediococcus acidilactici on growth performance,survival rate,and stress resistance of Persian sturgeon (Acipenser persicus)

This study was conducted to examine the effect of Artemia urmiana nauplii enriched with Pediococcus acidilactici on growth performances, survival rate, and stress resistance of Persian sturgeon (Acipenser persicus). Artemia nauplii were enriched with P. acidilactici at concentration of 10¹⁰ CFU/ml for 3 h (T3), 6 h (T6), 9 h (T9), and one nonenriched nauplii treatment (control). Since nauplii enriched for 9 h had the most significant CFU/g, fish were fed with T9 for 11 days and compared with the control. To evaluate the effect of probiotic on fish resistance, Persian sturgeon larvae were subjected to osmotic shocks of 15, 25, or 35 ppt; four pH treatments, pH 5, pH 6, pH 8 or pH = 9; and air for 5, 10, 15, or 20 s on the 11th day. There were no significant differences in final weight, weight gain, CF, SGR, and FCR between T9 and control (P > 0.05). However, the survival rate of larvae fed nauplii enriched for 9 h increased significantly in comparison with the control group (P < 0.05). In addition, significant higher resistance to all stress challenges was observed in the T9 group.     

Production of diatom–bacteria biofilm isolated from Seriola lalandi cultures for aquaculture application

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.     

Effect of combined use of Bacillus and molasses on microbial communities in shrimp cultural enclosure systems

To assess the effect of addition of probiotics and carbon source on the microbial community structure within the shrimp culture, an enclosure system simulating a high density, intensive aquaculture of Litopenaeus vannamei was set up in Dianbai, Maoming, Guangdong province, China. The phylogenetic profiles of microbial communities in the enclosure system after addition of Bacillus and molasses were studied in the middle and final culture season (on the 45th, 75th and 95th day) using polymerase chain reaction‐denaturing gradient gel electrophoresis. A total of four groups, each containing a triplicate were set up: Group A (control), Group B (64% of daily feed was provided as molasses with Bacillus sp.), Group C (50% of daily feed amount provided as molasses with Bacillus sp.) and Group D (64% of daily feed amount as molasses alone). The results showed that α‐Proteobacteria, δ‐Proteobacteria, Bacteroidetes and Actinobacteria were the dominant bacteria. Moreover, the relative proportion of α‐Proteobacteria increased and that of Bacteroidetes decreased gradually with time. Pathogenic Flexibacter was the dominant genus in Group A on the 45th day, while the functional strains (Sphingomonas sp., Erythrobacter sp., Roseobacter sp., and Rhodobacter sp.) were detected multiple times in Groups B and C. Bacillus sp. was the cultivable, dominant bacterium in Groups B and C on the 95th day. Therefore, the combined use of Bacillus and molasses helped increase diversity of the microbial community, effectively inhibit pathogens, and promote the formation and development of a beneficial microbial community structure in the high density, intensive aquaculture of L. vannamei.