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1.
To select and characterize potential probiotic bacteria from the gut microbiota of Siberian sturgeon (Acipenser baerii), 129 strains isolated from the hindgut were screened for antagonistic activity against five fish pathogens. Ten isolates showed antagonism towards three or more pathogens. Nine of these isolates were Gram‐positive, belonging to Lactococcus (seven) and Bacillus (two), and a single strain belonging to the Gram‐negative Citrobacter. These inhibitory isolates were identified using genetic, phentotypic and biochemical traits, and further characterized by in vitro tests assessing the adhesion and growth in mucus and resistance to gastric and intestinal fluids. The candidate probiotics were determined to be non‐pathogenic through an in vivo study. Based on these assays, Lactococcus lactis ssp. lactis STG45 and STG81 showed the broadest inhibitory potential, a high viability in simulated gastrointestinal juice and the highest adhesion capacity to mucus. They were therefore selected as the most promising candidate probiotics. This is the first study screening probiotics among the gut microflora of Siberian sturgeon.  相似文献   

2.
The autochthonous microbiota in the foregut, midgut and hindgut of juvenile grouper Epinephelus coioides following the dietary administration of probiotic Bacillus clausii for 60 days were assessed using polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE). A complex and generally similar bacterial composition along the digestive tract of E. coioides was detected in the DGGE profiles, while several bacteria showed regional specialization. Similarity dendrogram revealed that the bacterial composition of the foregut was more similar to the midgut than the hindgut. Samples collected from the probiotic group and the control group showed generally similar DGGE patterns, while no significant difference in the total number of bands and Shannon index were observed between the probiotic group and the control group, suggested that probiotic B. clausii exerted no significant effect on the gut microbiota of E. coioides. However, various potentially beneficial bacteria, such as Enterococcus sp.‐like and Bacillus pumilus‐like bacterium were selectively stimulated by probiotic B. clausii, while some potential harmful species, like Staphylococcus sp.‐like and Vibrio ponticus‐like bacterium were depressed. These indicated that the gut microbiota was modified to some degree by probiotic B. clausii. Sequences analysis showed that the autochthonous gut bacteria of E. coioides could be classified into four groups, i.e. Proteobacteria, Firmicutes, Actinobacteria and unclassified bacteria.  相似文献   

3.
Probiotics renowned as valuable microbes serve as a potential alternative to control diseases in aquaculture and are considered as an efficient and environment‐friendly approach to reduce the use of antibiotics. The present study aims at the isolation of putative probiotic bacteria from the intestinal tract of different fish species from the Doaba region of Punjab, India. In this study, isolated bacterial strains were characterized based on their morphological, biochemical and molecular characterization by 16S rRNA gene sequencing, followed by in vitro evaluation of different selection parameters described in FAO/WHO guidelines. A total of 169 different bacterial strains were isolated from the gastrointestinal tract of 52 different fish species. After in vitro evaluation, out of 169 bacterial strains only five bacteria (S3, S7, BDK2', BDK7 and BDK9) identified as Enterococcus and Bacillus species showed antagonistic activity against the fish pathogen Pseudomonas aeruginosa (MTCC 4 673). These isolates were screened based on their response towards bile tolerance, pH tolerance, adhesion and drug susceptibility to different antibiotic discs. And, the in vivo results indicated improved growth and survival against the infection (P. aeruginosa) after oral administration of the probiotics. The observations of in vitro and in vivo evaluation indicate that these isolated probiotic strains serve as effective probiotics and can be used as a novel and safe treatment to cure current issues in aquaculture.  相似文献   

4.
Enzymes synthesized by the gut microbiome are recognized as vital for assisting host metabolism. This study was conducted to screen and identify bacterial isolates with enzyme-producing abilities from the intestine of milkfish Chanos chanos. Overall, 114 isolates were collected, of which 58% showed clear zones indicating various enzyme activities via plate-based assays. Specifically, 65 isolates were amylolytic, 1 isolate was cellulolytic, and 18 isolates exhibited both activities. The two most promising isolates—CC8 and CC27—were further subjected to spectrophotometric enzyme assays to determine their amylase, cellulase and protease activities. Results showed that amylase, cellulase and protease activities (6.00 ± 0.01, 0.52 ± 0.02 and 18.62 ± 0.16 U mg?1 protein, respectively) were higher in CC8 than in CC27. Using 16S rRNA gene sequencing, the isolates were putatively identified as Vibrio sp. and Bacillus cereus respectively. At a suspension (mL) to feed (g) ratio of 3:25, dietary supplementation of putative probiotics was carried out to evaluate the growth performance, survival and enzymatic activities of juvenile Nile tilapia Oreochromis niloticus. Results revealed that probiotic-supplemented fish exhibited better growth performance—final average body weight, weight gain, specific growth rate and feed conversion ratio—along with increased intestinal enzymatic activities compared with the control (p < 0.05). Survival rates ranged from 80% to 90% and were statistically similar in all treatments (p > 0.05). This study concludes that the application of Vibrio sp. CC8 and Bacillus cereus CC27, supplemented either in monostrain or multistrain preparation, can promote growth and enzymatic activities in juvenile Nile tilapia.  相似文献   

5.
The aim of this study was to evaluate probiotic properties and the aflatoxin B1 adsorption ability of yeasts isolated from rainbow trout intestine and fish feed to assess their use in the formulation of feed additives. Growth at pH 2, bacterial pathogens inhibition, bacterial pathogens co‐aggregation, autoaggregation, homologous and heterologous inhibition against lactic acid bacteria were evaluated. Moreover, aflatoxin B1 adsorption was tested. All strains were able to maintain viable (107 cells/ml) at pH 2. All strains isolated from intestine were identified as Kazaschtania exigua, while strains isolated from feed were all identified as Debaryomyces hansenii. Kazaschtania exigua RC035 and RC037 showed the strongest antimicrobial activity while K. exigua RC037 and RC038 were the most efficient co‐aggregating bacterial pathogens. All strains exhibited strong autoaggregation. None of the tested yeast strains showed homologous inhibition towards other yeasts and heterologous inhibition towards lactic acid bacteria strains. Debaryomyces hansenii RC031 demonstrated aflatoxin B1 adsorption capacity (21%). The results of the present study indicate that select strains of Kazaschtania exigua and D. hansenii showed potential to improve the health of rainbow trout by inhibiting pathogens and binding AFB1 and their use as probiotics may improve the production of rainbow trout in aquaculture systems.  相似文献   

6.
Isolation and enumeration of amylase, cellulase and protease‐producing autochthonous bacteria in the proximal intestine (PI) and distal intestine (DI) of three species of Indian major carps, catla (Catla catla), mrigal (Cirrhinus mrigala) and rohu (Labeo rohita), were investigated using the conventional culture‐based technique. Population levels of amylolytic strains were the highest in the PI of catla and the lowest in the DI of rohu. The highest viable count of cellulase and protease‐producing bacteria was recorded in the DI and PI of mrigal respectively. Among the bacteria isolated, 10 strains (five from PI and five from DI) were selected as potent enzyme producers according to a quantitative enzyme assay. The chosen strains were further identified by 16S rRNA gene sequence analysis. The five strains isolated from catla showed high similarity to Citrobacter sp. clone W2, Enterobacter sp. JA24, Bacillus coagulans strain TR, uncultured bacterial clone Hel3bc04 and Bacillus cereus strain UST2006‐BC004. The four strains isolated from mrigal were most closely related to Bacillus sp. KCd2, uncultured bacterial clone Hel3bd09, B. cereus strain BU040901‐020 and Citrobacter freundii strain YRL11, while the strain isolated from rohu probably belonged to Bacillus sp. GV.  相似文献   

7.
In this study, a total of 98 lactic acid bacteria isolated from rainbow trout intestines were screened for their probiotic properties. The isolates were tested for their ability to inhibit growth of Vagococcus salmoninarum and Lactococcus garvieae. Based on in vitro antagonism, 10 isolates were selected and evaluated pathogenicity in rainbow trout. Isolates were further investigated for hydrophobicity, bile salts and acid tolerance. These isolates were able to survive low pH and high bile concentrations and showed good adherence characteristics. Isolates were characterized phenotypically, and then, 16S rRNA gene sequence analysis was used for confirmation. Selected strains were administered orally at 108 cfu/g feed, and fish were challenged with V. salmoninarum and L. garvieae. The fish fed with lactic acid bacteria supplemented diets did not improve protection against V. salmoninarum. However, administration of Lactococcus lactis subsp. lactis M17 2‐2 and Lactobacillus sakei 2‐3 resulted in a significant reduction in mortality due to L. garvieae when compared to the control fish. RPS values were calculated as 80 and 53% in fish fed with L. sakei 2‐3 and L. lactis subsp. lactis M17 2‐2, respectively. Our results suggest that these strains could provide an alternative for lactococcosis control in aquaculture.  相似文献   

8.
The disease outbreaks in aquaculture system of wetlands are the major cause of fish mortality. Among various bacterial septicaemic diseases, fish mortality caused by Acinetobacter spp. is recently reported in different fish species. Fish disease outbreak was investigated in a wetland of West Bengal, India to identify the aetiological factors involved. The moribund fish were examined and subjected to bacterial isolation. Two bacterial causative agents were identified as Acinetobacter junii and Acinetobacter pittii by biochemical characterization and 16S rRNA gene amplification. Both the isolates were oxidase‐negative, nitrate‐negative, catalase‐positive and indole‐negative. The molecular identification using 16S rRNA gene sequencing and phylogenetic tree analysis further confirmed the two Acinetobacter spp. with 97%–99% similarity. The antibiotic resistance patterns of these two bacteria revealed that both of them were resistant to β‐lactam, cefalexin, cephalothin, amoxyclav, cefuroxime, cefadroxil, clindamycin, vancomycin and penicillin. In addition, A. pittii was also resistant to other antibiotics of cephams group such as ceftazidime and cefotaxime. In the challenge experiment, both A. junii and A. pittii were found to be pathogenic with LD50 of 1.24 × 105 and 1.88 × 107 cfu/fish respectively. Histopathological examination of gill, liver and kidney revealed prominent changes supporting bacterial septicaemia. The investigation reports for the first time on concurrent infection by A. junii and multidrug‐resistant (MDR)‐A. pittii as emerging fish pathogens to cause severe mortality in Labeo catla and Hypophthalmichthys molitrix in a freshwater wetland.  相似文献   

9.
We evaluated the effects of Bacillus cereus, as an additive in water and feed, on the gut microbiota and immunological parameters of Nile tilapia (Oreochromis niloticus) fingerlings. Experiments were performed in tanks and net cages respectively. Experiment 1: Tilapia were housed in tanks for 42 days, and Bcereus was added to the water at 1.0 × 104 cfu mL?1 (Treatment 1) and 1.0 ×105 cfu mL?1 (Treatment 2) weekly. For the control, no probiotic was added. Experiment 2: Tilapia were housed in cages for 42 days, and the feed was supplemented with Bcereus at 1.0 × 107 cfu g?1 (Treatment 1) and 1.0 × 108 cfu g?1 (Treatment 2) weekly. For the control, no probiotic was added. Each treatment contained three replicates, with 50 male tilapias per replicate. The fish from the probiotic treatments in both tank and cage experiments had significantly higher serum lysozyme and peroxidase activities than the control. In the cage experiment, alkaline phosphatase and total superoxide dismutase activities in tilapia were significantly higher in probiotic treatments compared with the control. The results of polymerase chain reaction‐denaturing gradient gel electrophoresis showed that B. cereus supplementation in the feed and water affected the autochthonous gut bacteria community of tilapia and stimulated various potentially beneficial bacteria. Therefore, B. cereus, as a water or feed additive, could enhance the immune status and affect the gut microbiota of tilapia. Bacillus cereus was more effective as a feed supplement rather than a water additive for enhancing the immune status of tilapia.  相似文献   

10.
Streptococcus agalactiae infections in fish are predominantly caused by beta‐haemolytic strains of clonal complex (CC) 7, notably its namesake sequence type (ST) 7, or by non‐haemolytic strains of CC552, including the globally distributed ST260. In contrast, CC23, including its namesake ST23, has been associated with a wide homeothermic and poikilothermic host range, but never with fish. The aim of this study was to determine whether ST23 is virulent in fish and to identify genomic markers of fish adaptation of S. agalactiae. Intraperitoneal challenge of Nile tilapia, Oreochromis niloticus (Linnaeus), showed that ST260 is lethal at doses down to 10cfu per fish, whereas ST23 does not cause disease at 10cfu per fish. Comparison of the genome sequence of ST260 and ST23 with those of strains derived from fish, cattle and humans revealed the presence of genomic elements that are unique to subpopulations of S. agalactiae that have the ability to infect fish (CC7 and CC552). These loci occurred in clusters exhibiting typical signatures of mobile genetic elements. PCR‐based screening of a collection of isolates from multiple host species confirmed the association of selected genes with fish‐derived strains. Several fish‐associated genes encode proteins that potentially provide fitness in the aquatic environment.  相似文献   

11.
12.
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   

13.
In this study, 318 bacterial strains were isolated from the gastrointestinal (GI) tracts of 29 rainbow trout, Oncorhynchus mykiss (Walbaum). These bacteria were screened in vitro for their ability to inhibit growth of Flavobacterium psychrophilum, the causative agent of coldwater disease. Bacteria observed to inhibit F. psychrophilum growth were further screened against rainbow trout bile, as an indicator of their ability to survive in the GI tract. This screening resulted in narrowing the pool to 24 bacterial isolates. Those 24 isolates were then tested for pathogenicity in rainbow trout by intraperitoneal injection. Following a 28‐day challenge, eight isolates were shown to cause direct mortality and were eliminated from further study. As a result, 16 bacterial isolates were identified as probiotic candidates with the potential to control or reduce disease caused by F. psychrophilum.  相似文献   

14.
Carps are the most diversified freshwater fish belonging to family Cyprinidae. Numerous probiotic and pathogenic lactic acid bacteria (LAB) have been characterized from carps. However, the diversity of these ecologically important bacteria is entirely unknown in freshwater fish of Pakistan. The present study aimed to characterize and identify the lactic acid bacteria from two carps viz. Laboe rohita and Cirrhinus mrigala and determine their antagonistic activity. Seventeen bacterial isolates were purified from the gastrointestinal tract and gills of these fish and characterized morphologically. Initially, seven isolates were screened as LAB using agar supplemented with CaCO3. Subsequently, only two isolates CILB2 and RIL10 were selected as LAB after high‐performance liquid chromatography analysis for lactic acid production. Isolates CILB2 and RIL10 were genetically identified as Enterococcus faecalis and Weissella sp., respectively after 16S rRNA gene sequence analysis. Both strains exhibited significant antagonistic activity against common fish pathogens Streptococcus agalactiae, Staphylococcus aureus and Escherichia coli. Enterococcus faecalis CILB2 and Weissella sp. RIL10 were also found negative for haemolysis and gelatinase activities and were sensitive to ampicillin, amoxicillin, doxycycline, erythromycin, chloramphenicol and co‐trimoxazole antibiotics. The identified LAB strains may further be investigated for their potential probiotic application in fish feed and food preservation techniques.  相似文献   

15.
The effect of dietary β‐glucan on the bacterial community in the gut of common carp (Cyprinus carpio) was examined after oral application of Aeromonas hydrophila. Carp received either feed supplemented with 1% MacroGard®, a β‐1,3/1,6‐glucan, or a β‐glucan‐free diet. Fourteen days after feeding, half of the carp from each group were intubated with 109 colony‐forming units (CFU) of a pathogenic strain of A. hydrophila. Gut samples were taken 12 hr to 7 days after application and analysed using microbiological and molecular biological techniques (NGS, RT‐PCR‐DGGE). The reaction of the mucosa and the microbiota to an A. hydrophila intubation differed in carp fed with β‐glucan compared to carp from the control group. In β‐glucan fed carp, the total bacterial amount was lower but the number of bacterial species was higher. Bacterial composition was different for carp from both treatment groups. The number of mucin filled goblet cells was reduced in carp fed the β‐glucan diet. Mucus was obviously released from the goblet cells and was probably washed out of the gut together with high numbers of bacteria. This might be protective against pathogenic bacteria and, therefore, feeding with β‐glucan may provide protection against infections of the gut in carp.  相似文献   

16.
A study was conducted to characterize the autochthonous gut microbiota present in the pyloric caeca (PC), anterior mucosa (AM) and posterior mucosa (PM) of brown trout (Salmo trutta). Total viable counts (TVC) bacterial populations were enumerated using tryptone soy agar, lactic acid bacteria (LAB) levels were enumerated on de Man, Rogosa & Sharpe agar and PCR‐DGGE was employed as a culture‐independent method to assess the total communities. No significant differences were observed between the different gut regions for TVC or LAB levels. 16S rRNA sequencing identified all LAB isolates as Carnobacterium maltaromaticum. In contrast, the TVC community was more diverse; Firmicutes and Bacteroidetes were present but all gut regions were dominated by Proteobacteria, accounting for 88.4–92.6% of the communities. Citrobacter freundii was the dominant species and accounted for 51.0–57.8% of the isolates. Complex bacterial communities were observed using PCR‐DGGE and a trend towards the reduction in the number of operational taxonomic units (OTUs), microbial richness and microbial diversity was observed from the PC to the PM. The similarity between regions was low (52–68%) and cluster analysis revealed that the communities grouped into two distinct clusters; one dominated by the PM samples and the other contained the AM and PC samples. OTUs from the DGGE were identified as members of the phyla Proteobacteria and Firmicutes. Many OTUs were detected in all gastrointestinal regions, however, some OTUs showed regional specialization. Further studies are required to elucidate the activity of these genera in situ and how their actions impact the host.  相似文献   

17.
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   

18.
The purpose of the present study is to profile the microfloral structure in the intestines of the white shrimp, Litopenaeus vannamei, after being fed a control diet, in comparison with the diets containing Bacillus subtilis E20‐fermented soybean meal (FSBM) or an antimicrobial peptide (AMP) isolated from B. subtilis E20‐FSBM (diets designated FSBMD and AMPD, respectively) for 60 days. Metagenomic data showed detection of eight phyla, 20 classes, 40 orders, 68 families and 96 genera. Despite no statistically significant difference, an evaluation of microbial diversity recorded higher species richness, Shannon–Weaver diversity index and evenness in the AMPD and FSBMD groups, compared to the control diet group. Venn diagrams showed that 58 of the operational taxonomic units (OTUs) were shared among the control, FSBMD and AMPD groups, but additional beneficial microbes were only found in the FSBMD and AMPD groups. In addition to the augmentation of beneficial bacteria in the FSBMD and AMPD groups, the abundance of potential pathogens, Vibrio and Flavobacterium, was lower in the gut of shrimp fed the FSBMD and AMPD. The results clearly suggest that the FSBMD and AMPD have the capability to change the microfloral structure of shrimp intestines and could be used for disease prevention in shrimp aquaculture.  相似文献   

19.
Duplicate groups of Atlantic salmon (Salmo salar L.), kept in seawater, were fed fish meal‐based cold‐pelleted diets. Diets with non‐starch polysaccharides (NSP), either cellulose, purified soybean NSP or extruded purified soybean NSP at a dietary level of 100 g kg?1, were compared with a diet without supplemental NSP and a diet with soybean meal in a 28‐day feeding trial. Isolation and characterizations were limited to culturable bacteria and population levels of aerobic and facultative aerobic heterotrophic autochthonous (adherent) and allochthonous (transient) bacteria present in the mid and distal intestines of Atlantic salmon fed the five different diets estimated using traditional bacteriological techniques. The presence of an autochthonous microbiota was demonstrated using transmission electron microscopy. No significant effects of diet composition were observed on total population levels of culturable bacteria present in the digestive tract, but the study showed that the composition of the gut microbiota (autochthonous or allochthonous) was sensitive to dietary changes. A total of 752 culturable isolates from the intestines were characterized by biochemical and physiological properties. Of these, 188 isolates were further characterized by partial sequencing the 16S rRNA genes. Among these, 146 isolates belonged to 31 phylotypes that were >94% identical to previously described species. However, 42 isolates showed similarity <94% to species available at the National Center of Biotechnology Information. Several of the phylotypes identified in the present study have not been reported previously in the gastrointestinal (GI) tract of fish, including the Gram‐negative bacteria Gelidibacter salicanalis, Pseudoalteromonas elyakovii, Psychrobacter aquimaris, Psychrobacter cibarius, Psychrobacter fozii, Psychrobacter maritimus, Psychrobacter okhotskensis and Psychrobacter psychrophilus. Among the Gram‐positive bacteria identified were Arthrobacter bergeri, Arthrobacter psychrolactophilus, Arthrobacter rhombi, Bacillus pumilus, Bacillus subtilis, Exiguobacterium spp., Microbacterium oxydans, Planococcus maritimus, Sporosarcina ginsengisoli and several bacteria that have been described as unculturable previously. In addition, we identified Carnobacterium inhibens, a lactic acid bacterium that is not frequently isolated from the GI tract of fish. Psychrobacter cibarius was the dominant bacterial species and was isolated from the digestive tract of all fish investigated.  相似文献   

20.
The antibacterial effects of organic salts, chemical disinfectants and antibiotics were evaluated on cultures of Aeromonas hydrophila C03, Aeromonas sobria C26, A. sobria C29, Aeromonas caviae C24 and Acinetobacter sp. SH‐94B, the pathogens that cause black disease found in fairy shrimps (Streptocephalus sirindhornae Sanoamuang et al. (2000) and Branchinella thailandensis Sanoamuang, Saengphan & Murugan) of Thailand. The minimal inhibitory concentrations (MICs) of organic salts (sodium chloride and potassium chloride) and antibiotics (oxytetracycline dihydrate, streptomycin sulphate, kanamycin monosulphate, chloramphenicol and ampicillin) were determined using the agar‐dilution method. The effect of chemical disinfectants (sodium hypochlorite and chlorine dioxide) was evaluated by exposing bacteria to different concentrations of these chemicals for different periods of time. Interestingly, all strains were intrinsically resistant to 0.25–3% sodium chloride and potassium chloride. The effect of sodium hypochlorite was greater than that of chlorine dioxide, and 5–20 μg mL?1 of sodium hypochlorite was sufficient to inhibit the growth of these bacteria, but the exposure time varied, depending on the bacterial species. Of the antibiotics tested, chloramphenicol and oxytetracycline dihydrate completely inhibited the selected strains. Chloramphenicol showed the highest antibacterial effect against all pathogenic species – the MIC and minimal bactericidal concentration (MBC) ranged from 0.8 to 3.1 μg mL?1 from 3.1 to 6.25 μg mL?1, respectively. To achieve control of black disease during cultivation of fairy shrimp, data derived from this study can be used as a basis for further toxicity tests in vivo.  相似文献   

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