Bacteriological analysis of the digestive tube of the mud snail (Bullacta exarata Philippi) and its rearing shoal

The bacterial flora in the digestive tube of Bullacta exarata Philippi and its rearing shoal were investigated. A total of 157 strains of heterotrophic bacteria, isolated from crop, stomach intestine and other parts of the digestive tube, mainly belong to the genera Photobacterium, Bacillus, Pseudomonas Vibrio and some genera of the family Enterobacteriaceae. There are significantly more varieties of bacteria in crop than in stomach and intestine. A total of 173 strains of bacteria were isolated from the rearing shoal, belonging to 13 genera. The 5 predominant genera, such as Bacillus and Photobacterium, are the same as those in the digestive tube, but greatly differ in percentages. The number of heterotrophic bacteria and Vibrio in rearing shoal change in line with the alteration of the temperature, and are significantly affected by the use of pesticides.     

马氏珠母贝肠道及其养殖水体可培养细菌群落结构

【目的】了解马氏珠母贝(Pinctada fucata martensii)肠道及其养殖水体可培养细菌的群落组成。【方法】采用2216E平板涂布法研究海区养殖马氏珠母贝肠道与养殖水体的可培养菌群种类及丰度。【结果与结论】马氏珠母贝肠道及其养殖水体的可培养细菌归属于2门(变形菌门和厚壁菌门)3纲7目10科23属56种。属水平上,肠道中以弧菌属(74.7%)和假交替单胞菌属(18.7%)为主;养殖水体中α-变形菌纲的FJ943236_g属(40.7%)丰度最大,弧菌属(16.7%)相对肠道丰度较低。样品共有菌属为弧菌属、假交替单胞菌属、发光杆菌属和芽孢杆菌属;肠道特有菌属为希瓦氏菌属和盐单胞菌属;养殖水体特有菌属主要为FJ943236_g、鲁杰氏菌属和Nautella。在种水平上,7个种为二者共有;马氏珠母贝肠道和养殖水体特异性菌种分别为18个和31个。虽然门水平上马氏珠母贝肠道中可培养细菌群落与其养殖水体中的细菌群落大致相似,但在属、种水平上二者差异明显。     

具有多种胞外酶的对虾肠道黏附菌的分离和鉴定

用对虾饲料培养基从健康凡纳滨对虾肠道分离出500株黏附细菌,以产淀粉酶、脂肪酶和蛋白酶能力为指标,筛选出产该3种消化酶的细菌90株,占总菌株的18%.对其中生长较快的69株进行16S rDNA 基因测序,确定其分类地位.结果显示,69株菌分别属于不动杆菌属(Acinetobacter)、芽孢杆菌属(Bacillus)、葡萄球菌属(Staphylococcus)、假交替单胞菌属(Pseudoalteromonas)、气单胞菌属(Aeromonas)、嗜盐单胞菌属(Halomonas)、利斯顿氏菌属(Listonella)、莫拉氏菌属(Moraxella)等,其中数量最多是芽胞杆菌属,占鉴定细菌总数的53.62%,数量最少是气单胞菌属和嗜盐单胞菌属,均占鉴定细菌总数的2.90%.表明对虾肠道黏附菌群中具有较多能分泌多种消化酶的细菌,可进一步开发为促进对虾消化功能的益生菌     

Contribution of trimethylamine N-oxide on the growth and pressure tolerance of deep-sea bacteria

Trimethylamine N-oxide (TMAO) is widely dispersed in marine environments and plays an important role in the biogeochemical cycle of nitrogen. Diverse marine bacteria utilize TMAO as carbon and nitrogen sources or as electron acceptor in anaerobic respiration. Alteration of respiratory component according to the pressure is a common trait of deep-sea bacteria. Deep-sea bacteria from different genera harbor high hydrostatic pressure (HHP) inducible TMAO reductases that are assumed to be constitutively expressed in the deep-sea piezosphere and facilitating quick reaction to TMAO released from fish which is a potential nutrient for bacterial growth. However, whether deep-sea bacteria universally employ this strategy remains unknown. In this study, 237 bacterial strains affiliated to 23 genera of Proteobacteria, Bacteroidetes, Firmicutes and Actinobacteria were isolated from seawater, sediment or amphipods collected at different depths. The pressure tolerance and the utilization of TMAO were examined in 74 strains. The results demonstrated no apparent correlation between the depth where the bacteria inhabit and their pressure tolerance, regarding to our samples. Several deep-sea strains from the genera of Alteromonas, Halomonas, Marinobacter, Photobacterium, and Vibrio showed capacity of TMAO utilization, but none of the isolated Acinebacter, Bacillus, Brevundimonas, Muricauda, Novosphingobium, Rheinheimera, Sphingobium and Stenotrophomonas did, indicating the utilization of TMAO is a species-specific feature. Furthermore, we noticed that the ability of TMAO utilization varied among strains of the same species. TMAO has greater impact on the growth of deep-sea isolates of Vibrio neocaledonicus than shallow-water isolates. Taken together, the results describe for the first time the TMAO utilization in deep-sea bacterial strains, and expand our understanding of the physiological characteristic of marine bacteria.

     

Analysis of cultivable aerobic bacterial community composition and screening for facultative sulfate-reducing bacteria in marine corrosive steel

Anaerobic, aerobic, and facultative bacteria are all present in corrosive environments. However,as previous studies to address corrosion in the marine environment have largely focused on anaerobic bacteria, limited attention has been paid to the composition and function of aerobic and facultative bacteria in this process. For analysis in this study, ten samples were collected from rust layers on steel plates that had been immersed in seawater for dif ferent periods(i.e., six months and eight years) at Sanya and Xiamen,China. The cultivable aerobic bacterial community structure as well as the number of sulfate-reducing bacteria(SRB) were analyzed in both cases, while the proportion of facultative SRB among the isolated aerobic bacteria in each sample was also evaluated using a novel approach. Bacterial abundance results show that the proportions are related to sea location and immersion time; abundances of culturable aerobic bacteria(CAB) and SRB from Sanya were greater in most corrosion samples than those from Xiamen,and abundances of both bacterial groups were greater in samples immersed for six months than for eight years. A total of 213 isolates were obtained from all samples in terms of CAB community composition,and a phylogenetic analysis revealed that the taxa comprised four phyla and 31 genera. Bacterial species composition is related to marine location; the results show that Firmicutes and Proteobacteria were the dominant phyla, accounting for 98.13% of the total, while Bacillus and Vibrio were the dominant genera,accounting for 53.06% of the total. An additional six facultative SRB strains were also screened from the isolates obtained and were found to encompass the genus Vibrio(four strains), Staphylococcus(one strain),and Photobacterium(one strain). It is noteworthy that mentions of Photobacterium species have so far been absent from the literature, both in terms of its membership of the SRB group and its relationship to corrosion.     

Effects of a Potential Autochthonous Probiotic Bacillus subtilis 2-1 on the Growth and Intestinal Microbiota of Juvenile Sea Cucumber,Apostichopus japonicus Selenka

The effects of Bacillus subtilis 2-1 from the intestine of healthy sea cucumber on the growth, digestive enzyme activities and intestinal microbiota of juvenile sea cucumber(Apostichopus japonicus) were determined in the present study. Sea cucumber was fed with Sargassum thunbergii powder supplemented with B. subtilis 2-1 at different concentrations varying among 0(control), 10~5, 10~7, and 10~9 CFU g~(-1) for 8 weeks. Results showed that the growth performance and intestinal amylase and trypsin activities were significantly increased by dietary B. subtilis 2-1 at 10~9 CFU g~(-1)(P 0.05). However, dietary B. subtilis 2-1 had no significant influence on the lipase activity in sea cucumber(P 0.05). The polymerase chain reaction denaturing gradient gel electrophoresis and 16S rRNA gene sequencing analysis indicated that dietary B. subtilis 2-1 at 105 and 107 CFU g~(-1) inhibited most of the Proteobacteria including those in genus Vibrio. Dietary B. subtilis 2-1 at 10~9 CFU g~(-1) not only decreased the abundance and species of genus Vibrio, but also increased the intensity of genera Psychrobacter and Bacillus. A specific dosage of dietary B. subtilis 2-1 could increase the growth and modulate the intestinal microbiota of sea cucumber; thus it might be a novel probiotic for keeping the health of sea cucumber.     

Diversity of bacteria isolated from crustacea larvae and their rearing water

1　Introduction　Blacktigershrimp (Penaeusmonodon)isthemajorshrimpspeciesculturedinIndonesia .Itsculturehasbecomemoreintensivebecauseofitshighdemandandeconomicvalue .Infact,shrimpexporthasyieldedthehighestIndonesianincomeinfisherysector .Itwasestimatedthattheproductionofculturedshrimpwas 80 0 0 0metrictonnesin 1995 .Consequently ,theneedforthefryforstockingpondculturesteadilyin creased .Thetotalnumberoffryrequiredwasabout 6millionperyear (FAO ,1995 )and 90 %oftherequiredanimalswereobtained…     

Structure and Function of the Digestive System of Solen Grandis Dunker

Structure and function of the digestive system of a bivalve mollusc, Solen grandis, were studied using light microscopy and histochemical methods. The wall of digestive tube consists of four layers: the mucosal epithelium, connective tissue, muscular and fibrosa or serosa (only in the portion of rectum) from the inner to the outer. The ciliated columnar epithelial cells, dispersed by cup-shaped mucous cells, rest on a thin base membrane. There are abundant blood spaces in connective tissue layer. The digestive diverticula are composed of multi-branched duct and digestive tubules. The digestive tubules are lined with digestive and basophilic secretory cells, and surrounded by a layer of smooth muscle fibers and connective tissues. Activities of acid and alkaline phosphatases, esterase and lipase are detected in the digestive cells, and the epithelia of stomach and intestine, suggesting that these cells are capable of intracellular digesting of food materials and absorbing. Besides, acid phosphatase and e     

Structure and function of the digestive system of <Emphasis Type="Italic">solen grandis</Emphasis> dunker

Structure and function of the digestive system of a bivalve mollusc, Solen grandis, were studied using light microscopy and histochemical methods. The wall of digestive tube consists of four layers: the mucosal epithelium, connective tissue, muscular and fibrosa or serosa (only in the portion of rectum) from the inner to the outer. The ciliated columnar epithelial cells, dispersed by cup-shaped mucous cells, rest on a thin base membrane. There are abundant blood spaces in connective tissue layer. The digestive diverticula are composed of multi-branched duct and digestive tubules. The digestive tubules are lined with digestive and basophilic secretory cells, and surrounded by a layer of smooth muscle fibers and connective tissues. Activities of acid and alkaline phosphatases, esterase and lipase are detected in the digestive cells, and the epithelia of stomach and intestine, suggesting that these cells are capable of intracellular digesting of food materials and absorbing. Besides, acid phosphatase and esterase activities are present in the posterior portion of esophagus. Phagocytes are abundant in blood spaces and the lumens of stomach and intestine, containing brown granules derived from the engulfed food materials. The present work indicates that phagocytes play important roles in ingestion and digestion of food materials, which is supported as well by the activities of acid phosphatase, esterase and lipase detected in blood spaces.     

Histological Study on the Digestive System Development of Takifugu rubripes Larvae and Juvenile

The digestive tract of Takifugu rubripes during early life stages was studied with light microscopy. At the beginning of hatching, the digestive tract is represented by a simple and undifferentiated straight tube and does not communicate with the exterior, as the mouth and anus are not open yet. At 2 d after hatching, a constriction between intestine and rectum that will become the intestino-rectal valve is visible. During the endogenous feeding phase, the yolk sac is resorbed and the digestive tract becomes functional and differentiated. The liver and pancreas also become apparent at this time. At onset of exogenous feeding （3 d after hatching）, yolk sac resesves are not completely depleted, suggesting a period of mixed nutrition. The digestive tract differentiates fully into b.uccopharynx, esophagus, stomach, intestine and rectum. At 9 d after hatching, the yolk sac reserves are completely depleted. The most noticeable events occurring from 5 to 17 d after hatching are the transformation of the epithelia type, the differentiation of the pneumatic sac, the epithelial cell, gut convolution, mucosal fold increase, and the growth of liver and pancreas. From 18 to 27 d after hatching, the numbers of intestine folds and mucus cells increase progressively. From then on, morphological changes of digestive features are almost completed. At 27 d after hatching, the morphology and function of digestive system are similar to that of the adult.     

Heterotrophic bacterial flora in aquaculture area around Xuejiadao

From Oct., 1999 to Oct., 2000, the heterotrophic bacterial flora in the aquaculture area around Xuejiadao was investigated. The result shows that the populations of the heterotrophic bacteria are heavier in summer and autumn than those in winter and spring. The average populations in seawater, sediment, the surface of seaweed and the surface of fish are 1.4×10⁴cfu mL⁻¹, 5.4×10⁶cfu g⁻¹, 1.5×10⁶cfu g⁻¹ and 1.8×10³cfu cm⁻², respectively. A total of 301 strains were isolated, among them 259 were Gram-negative. All the Gram-negative bacteria belong to 13 genera and some genera of Enterobacteriaceae. The communities of bacteria are slightly different among the samples. In the body surface of fish, Genus vibrio is dominant. In the remaining samples, dominant genus is Aeromonas.     

Heterotrophic bacterial floras in industrial area and unpolluted marine environments around Qingdao

From Oct. 1999 to Oct. 2000, the heterotrophic bacterial floras in the industrial marine environment around the Qingdao Power Plant (QPP) and in the unpolluted marine environments were investigated. The results showed that the numbers of the heterotrophic bacteria around QPP were much higher than those in unpolluted environments, and the average numbers in QPP Seawater, QPP Sediment, Unpolluted Seawater and Unpolluted Sediment were 5.4×10⁴cfu(mL)⁻¹, 5.0×10⁵cfug⁻¹, 3.0×10²cfu(mL)⁻¹ and 1.3×10⁵cfug⁻¹ respectively. Totally, 118 strains were isolated from QPP and 99 of them were Gram-negative. One hundred and twenty one strains were isolated from the unpolluted environments and 104 of them were Gram-negative. All the Gram-negative bacteria belonged to 13 genera. The distribution of the bacteria was varied in different marine environments. The results showed that the unpolluted marine environments contained much more Vibrio than seawater and sediment around QPP.     

Isolation and identification of bacteria associated with the surfaces of several algal species

We conducted this study to assess the diversity of bacteria associated with the surfaces of algae based on 16S rDNA sequence analyses.Twelve strains of bacteria were obtained from the surfaces of the following four species of algae:Gracilaria textorii,Ulva pertusa,Laminaria japonica,and Polysiphonia urceolata.The isolated strains of bacteria can be divided into two groups:Halomonas and Vibrio,in physiology,biochemical characteristics and 16S rDNA sequence analyses.The phylogenetic tree constructed based on ...     

Isolation and characterization of AHL-degrading bacteria from fish and pond sediment

Quorum sensing(QS)disruption is considered as a potential alternative strategy to combat bacterial diseases in aquaculture.In this study,we isolated and identified bacteria degrading QS molecules from pond sediment and fish intestine.A total of 132 strains were obtained in the enrichment culture,of which two strains were identified as Enterobacter sp.f003 and Staphylococcus sp.sw120,being isolated from the fish intestine and pond sediment,respectively.We found that strains f003 and sw120 could degrade acyl-homoserine lactones(AHLs)and cause no hemolysis of sheep red blood cells.The AHL lactonase(aiiA)homologous gene in the two strains was detected in PCR amplification and the high-degrading activity to N-hexanoyl-L-homoserine lactone(C6-HSL)and AHLs secreted from pathogenic Aeromonas hydrophila was assessed.Meanwhile,the artificial infection of cyprinid Carassius auratus gibelio with intraperitoneal injection showed that the two strains were avirulent.Therefore,the obtained indigenous bacteria are candidate probiotics against pathogenic A.hydrophila in aquaculture.     

饥饿和再投喂对尼罗罗非鱼蛋白酶活性的影响

测定了饥饿和再投喂的尼罗罗非鱼胃、肠、肝胰脏蛋白酶活性。结果表明:(1)尼罗罗非鱼蛋白酶的活性以胃为最高,肠为最低,且胃蛋白酶的活性极显著高于肠、肝胰脏(P<0.01),肠和肝胰脏之间差异不显著(P>0.05);(2)饥饿后,蛋白酶活性呈降低的趋势,饥饿处理25d后,胃、肠、肝胰脏的蛋白酶活力分别降低到饥饿处理前的42.7%、41.0%、61.5%,均与对照组差异极显著(P<0.01);(3)再投喂后,饥俄15d以内的鱼蛋白酶活性上升迅速,经过15d恢复投喂,基本上就恢复到了正常水平,且消化酶活性甚至高于一直投喂对照组,而饥饿20、25d的鱼蛋白酶活性虽缓慢上升,但仍无法恢复到正常水平,与对照组差异显著(P<0.05)。     

Influence of Probiotics on the Growth and Digestive Enzyme Activity of White Pacific Shrimp （Litopenaeus vannamel）

The influence of Bacillus probiotics on the digestive enzyme activity and the growth of Litopenaeus vannamei were de-termined in this study. The shrimp was treated with five percentages (1.5, 3.0, 4.5, 6.0 and 7.5) of probiotics (Bacillus spp.) supple-mented to the feed and cultured for 45d. The growth measured as the weight gain at the end of culturing was significantly (P<0.05) higher in probiotic-treated shrimps than that of the control (without receiving probiotics). Activities of protease and amylase, two digestive enzymes of the midgut gland and the intestine were significantly (P<0.05) higher in probiotic-treated shrimp than in the control.     

近江牡蛎肠道细菌及其产酶能力

从近江牡蛎肠道中分离出21株菌,其中, 9株为革兰阳性菌, 12株革兰阴性菌。为寻找能分泌多种酶的有益微生物,研究了它们蛋白酶、脂肪酶、淀粉酶、纤维素酶的产酶能力。结果表明,有61 .9%的菌株能分泌蛋白酶或淀粉酶(各有13株菌);有52. 4%的菌株能分泌脂肪酶(11株菌);有33. 3%的菌株能产纤维素酶(7株菌)。在这21株菌中,产4种酶的有5株菌,产3种酶的3株菌,产两种酶的5株菌,产1种酶的3株菌,不产酶的仅有5株菌。不产酶的菌株仅占细菌总数的23. 8%,而各种产酶菌则高达76. 2%,由此表明细菌在近江牡蛎对食饵消化过程的重要性。     

Comparative analysis on microbial community associated with different gastrointestinal regions of wild northern snakehead Channa argus Cantor, 1842

Microbial communities in different gastrointestinal regions(stomach, foregut, midgut, and hindgut) of the northern snakehead C hanna argus(Cantor, 1842) were compared by polymerase chain reaction and partial 16 S r DNA sequencing. A total of 194, 140, 212, and 122 OTUs were detected in the stomach, foregut, midgut, and hindgut, respectively. Significant dif ferences were found in the Sobs, ACE, Shannon, and Simpson indices among samples(P 0.05). The gastrointestinal microbial community of C. argus consisted predominantly of Proteobacteria with either H alomonas, Shewanella, Plesiomonas, or Sphingomonas. Fusobacteria, Firmicutes, and Bacteroidetes also existed in the gastrointestinal tracts. However, significant diff erences were found in the compositions of microbial community among the four regions( P 0.05). Cyanobacteria and Spirochetes were significantly higher in the midgut and hindgut( P 0.05). Fusobacteria and Firmicutes were dominant in the hindgut and foregut, respectively(P 0.05). Proteobacteria was the lowest in the hindgut( P 0.05). At genus level, C etobacterium and Plesiomonas were significantly higher in the hindgut than in the other three samples( P 0.05). Clostridium and Prevotella were the highest in the midgut(P 0.05). Halomonas, Shewanella, and S phingomonas were the highest in the foregut( P 0.05). Paracoccus and Vibrio were the highest in the stomach. Several genera were only detected in certain regions, as follows: stomach, P aracoccus and Vibrio; foregut, Halomonas, Shewanella, and Sphingomonas; midgut, Clostridium and Prevotella; and hindgut, C etobacterium and Plesiomonas( P 0.05). At the species level, A cinetobacter rhizosphaerae was only detected in the stomach. P revotella copri and Clostridium perfring were not detected in the foregut and midgut, respectively, whereas Prevotella copri and Faecalibacterium pra were not detected in the hindgut. These findings provide valuable information on the microbial community in each gastrointestinal region of C. argus. Moreover, this study indicated that microbial community was not only related to rearing environment but also to the physico-chemical characteristics of each gastrointestinal region.     

Digestive enzyme activities in mudskipper Boleophthalmus pectinirostris and Chinese black sleeper Bostrichthys sinensis

The mudskipper Boleophthalmus pectinirostris and Chinese black sleeper Bostrichthys sinensis occupy the intertidal zone. However, both species have their own unique diet. The former is an herbivore and the latter is a carnivore. In order to reveal the relationship between digestive enzyme activities and diets in the two species, the activities of protease (P), non-specific bile salt-activated lipase (BAL) and α-amylase (A) were determined in the stomach and intestine of adult mudskipper B. pectinirostris and Chinese black sleeper B. sinensis. The results showed that the activities of protease, BAL and α-amylase in the intestine of B. pectinirostris were significantly (P<0.05) higher than those in the stomach. In B. sinensis, gastric protease activity was not different from the intestinal protease (P>0.05), while BAL and α-amylase activities of the intestine were significantly (P<0.05) higher than those of the stomach. The activity of gastric protease in B. sinensis was significantly (P<0.05) higher than that in B. pectinirostris, while the activities of intestinal protease were not different between the two fish species (P>0.05). BAL activities of the stomach and intestine in B. sinensis were significantly (P<0.05) higher than those in B. pectinirostris, while α-amylase activities of the stomach and intestine in B. pectinirostris were significantly (P<0.05) higher than those in B. sinensis. The ratios of P/BAL, A/P and A/BAL of the digestive tract in B. pectinirostris were 1.5, 107.3 and 158.6, respectively; and those in B. sinensis were 0.2, 1.6 and 0.2, respectively. It can be concluded that food digestion in the adult B. pectinirostris is mainly carried out in the intestine, whereas in the adult B. sinensis it is initiated in the stomach and finishes in the intestine. The activities of BAL and α-amylase in B. pectinirostris and B. sinensis are well correlated with their diets. However, a clear-cut correlation between protease activity and diets is not found in these two species.     

Fucoxanthin Isolated from Undaria pinnatifida Can Interact with Escherichia coli and lactobacilli in the Intestine and Inhibit the Growth of Pathogenic Bacteria

Fucoxanthin is a xanthophyll-type carotenoid that provides many benefits to human health. However, the mechanism by which fucoxanthin interacts with microbes and inhibits pathogenic bacteria is unknown. In this study, we investigated the effects of fucoxanthin isolated from the edible seaweed Undaria pinnatifida on pathogenic bacteria Escherichia coli and lactobacilli both in vitro and in vivo. Fucoxanthin strongly inhibited the growth of Gram-positive pathogenic bacteria but was less effective against Gram-negative bacteria. Fucoxanthin extracted from the crude mixture had a recovery rate of 93.38% and a purity of 82.70%, which were higher than those of fucoxanthin extracted using a previous method. Fucoxanthin also promoted the growth of intestinal microbes in mice. Fucoxanthinol, a metabolite of fucoxanthin, was generated in the culture media. Fucoxanthin can be deacetylated into fucoxanthinol not only by conventional digestive enzymes in the digestive tract, but also by E. coli and lactobacilli in the intestine. These results indicate that fucoxanthin interacts with and influences E. coli and lactobacilli in the intestine. Therefore, fucoxanthin isolated from Undaria pinnatifida possibly can be applied in human health maintenance.