16S rRNA metagenomic analysis of the symbiotic community structures of bacteria in foregut,midgut, and hindgut of the wood-feeding termite <Emphasis Type="Italic">Bulbitermes</Emphasis> sp.

The termite gut is a highly structured microhabitat with physicochemically distinct regions. It is generally separated into the foregut, midgut and hindgut. The distribution of gut microbiota is greatly influenced by varying physicochemical conditions within the gut. Thus, each gut compartment has a unique microbial population structure. In this study, the bacterial communities of foregut, midgut and hindgut of wood-feeding higher termite, Bulbitermes sp. were analyzed in detail via metagenomic sequencing of the 16S rRNA V3-V4 region. While the microbiomes of the foregut and midgut shared a similar taxonomic pattern, the hindgut possessed more diverse bacterial phylotypes. The communities in the foregut and midgut were dominated by members of the group Bacilli and Clostridia (Firmicutes) as well as taxon Actinomycetales (Actinobacteria). The main bacterial lineage found in hindgut was Spirochaetaceae (Spirochaetes). The significant difference among the three guts was the relative abundance of the potential lignin-degrading bacteria, Actinomycetales, in both the foregut and midgut. This suggests that lignin modification was probably held in the anterior part of termite gut. Predictive functional profiles of the metagenomes using 16S rRNA marker gene showed that cell motility, energy metabolism and metabolism of cofactors and vitamins were found predominantly in hindgut microbiota, whereas xenobiotics degradation and metabolism mostly occurred in the foregut segment. This was compatible with our 16S rRNA metagenomic results showing that the lignocellulose degradation process was initiated by lignin disruption, increasing the accessibility of celluloses and hemicelluloses.     

草鱼肠道黏膜厌氧细菌的分离与鉴定

研究以草鱼(Ctenopharyngodon idellus)为实验对象, 运用厌氧培养的方法, 研究了饥饿状态下草鱼肠道黏膜固有微生物的类群及其在不同肠段的分布。实验结果显示草鱼前肠、中肠与后肠中细菌的数量分别是3.17×103、1.63×104和1.79×107 cfu/g。研究共分离到274株单菌落, 经16S rRNA鉴定, 分别属于拟杆菌属(Bacteroides spp.)、鲸杆菌属(Cetobacterium spp.)、梭形杆菌属(Fusobacterium spp.)、气单胞菌属(Aeromonas spp.)、希瓦氏菌属(Shewanella spp.)、芽孢杆菌属(Bacillus spp.)、泛菌属(Pantoea spp.)和柠檬酸杆菌属(Citrobacter spp.)8个种类, 其中专性厌氧细菌的数量占9.1%, 兼性厌氧细菌的数量占90.9%。进一步分析发现, 前肠中只分离到兼性厌氧细菌, 中肠与后肠专性厌氧细菌和兼性厌氧细菌都有分布。专性厌氧细菌Bacteroides paurosaccharolyticus和Bacteroides luti在中肠与后肠都有分布, 而Cetobacterium somerae和Fusobacterium ulcerans只在后肠有发现。兼性厌氧细菌是草鱼肠道黏膜的优势菌群, 其中嗜水气单胞菌Aeromonas hydrophila占73.7%。草鱼肠道不同部位固有厌氧微生物组成存在差异, 细菌数量也明显不同, 后肠中具有更高的细菌丰度和多样性。     

Immunofluorescence assay for effects on field abundance of a naturally occurring pseudomonad during passage through the gut of a marine deposit feeder,Abarenicola pacifica

In a seasonal study we used immunofluorescence to follow a specific bacterial population, as well as total numbers, through the fore-, mid-, and hindgut of a deposit feeder, Abarenicola pacifica. We chose a pseudomonad because of its high ambient abundance. On five dates, we collected A. pacifica gut contents, with concurrent measurements of sedimentary food quality (chlorophyll a, protein, bacterial abundance), animal egestion rates (inversely proportional to gut residence time), and temperature. Increasing bacterial numbers from ingested sediment to foregut contents, and decreases from foregut to midgut indicate significant selection and digestion, respectively, of both the pseudomonad and the total bacterial community. Inverse correlations between egestion rate and digestive removal of bacteria offer some support for the prediction that digestion of bacteria is proportional to time spent exposed to digestive enzymes, although the significance of the associated statistical tests is marginal. No hindgut growth of the pseudomonad was observed, likely due to the short gut residence time of A. pacifica. The pseudomonad showed variation of less than a factor of 3 in its ambient sedimentary abundance over the year. Off print requests to: C. Plante.     

The functional morphology of the alimentary canal of larval stages of the parasitic copepod Lepeophtheirus salmonis

The functional morphology of the alimentary canal of copepodite and chalimus stages of Lepeophtheirus salmonis (Krøyer, 1837) is described and compared with that found in other copepods studied to date.
The buccal cavity passes into a gut comprising three major regions: foregut (oesophagus), midgut and hindgut. The foregut and hindgut both posscss a cuticular lining whereas the midgut is lined with specialized epithelial cells. The midgut is divided into three recognizable zones, namely anterior midgut caecum, anterior midgut and posterior midgut. Three main types of epithelial cell are recognizable in the midgut: vesicular cells, microvillous cells and basal cells which correspond to the cell types normally described in other parasitic and free-living copepod species.
Digestion is thought to occur in the midgut and be mediated by the epithelial cells that line it. Although several glands appear to discharge into the area of the buccal cavity, none was seen to interface to any other area of the gut. There was no evidence for the involvement of commensal gut bacteria in food digestion.     

The origin and distribution of cellulase in the termites,Nasutitermes exitiosus and Coptotermes lacteus

The cellulase of the higher termite Nasutitermes exitiosus was located in the foregut (19%), the midgut (59%), the mixed segment (14%) and in the hindgut (8%). Removal of the gut flora by feeding tetracycline or by starving the termite did not affect the activity of the enzymes indicating that the termite secretes its own cellulase and is not dependent on its gut flora for the digestion of cellulose. The cellulase of the lower termite Coptotermes lacteus was distributed through the foregut (19%), midgut (32%) and hindgut (49%). Removal of the gut flora and fauna, left the specific activity of the cellulase of the fore- and mid-gut largely unaffected, but led to a 20% decrease in the specific activity of the cellulase in the hindgut. In starved C. lacteus the distribution of cellulase activity was foregut, 44%; midgut, 30% and hindgut, 26%. These results indicate that C. lacteus synthesises its own cellulase in addition to using the gut protozoa for cellulose digestion.     

Carbohydrases and carbohydrate digestion in the gut of Locusta migratoria

The digestion of various carbohydrates and synthetic substrates by the gut of Locusta migratoria was analysed quantitatively. Maltose, starch, and sucrose were found to be hydrolysed most rapidly, whereas the splitting of cellobiose, trehalose, lactose, and melecitose took place at much slower rates.The absolute carbohydrase activities in foregut and midgut are nearly equal. However, specific activities are much higher in the foregut. Only low activities were found in extracts from the hindgut and salivary glands. The latter show a pattern of sugar splitting which is different from that found in gut preparations.The distribution of carbohydrase activities between the epithelia and lumina of the foregut, midgut, and hindgut and between soluble and particulate fractions were studied. The midgut epithelium is shown to have a particularly high content of enzymes, although some carbohydrases are rather active also in the epithelium of the hindgut. During hunger periods the relative enzymatic activities of the epithelium are distinctly increased.The isolation and purification of the carbohydrases were attempted and a partial separation of individual enzymes was obtained by gel-filtration. These results indicate the presence of at least seven distinct carbohydrases in the locust gut. The molecular weights of the enzymes were estimated by gel-filtration, and K_M values and pH-optima are reported.     

Abundance and distribution of the gut flora of the desert locust,Schistocerca gregaria

The abundance of bacteria in the gut of Schistocerca gregaria was determined. A large population of Enterobacteriaceae was found with numbers increasing posteriorly from foregut to rectal sac. In the hindgut, Enterobacter agglomerans was the dominant organism. Streptococci were also present but they were 10- to 100-fold less numerous than the Enterobacteriaceae. The distribution of the microflora was investigated using light microscopy and scanning electron microscopy. Bacteria in the anterior regions of the gut were restricted to the lumen and inside of the peritrophic membrane. However, in the hindgut, bacteria were also associated with the cuticular lining.     

Lactic acid bacteria associated with the digestive tract of Atlantic salmon (Salmo salar L.)

The present study reports the effect of excessive handling stress and starvation on the lactic acid bacteria associated with the digestive tract of Atlantic salmon (Salmo salar L.). A relatively low population level (approximately 2 x 103 bacteria per gram wet tissue) of viable adherent heterotrophic bacteria was associated with the digestive tract (foregut, midgut and hindgut). Of the 752 bacterial isolates isolated from diet, water and the digestive tract, 201 isolates belonged to the carnobacteria. Of these isolates, one from the diet, one from the rearing water and 80 from the gastrointestinal tract, were further identified on the basis of 16S rDNA sequence analysis. All these isolates were identified as being Carnobacterium piscicola-like. Daily repeated stress and starvation of the fish over 11 d had no influence on the total culturable bacterial numbers or population level of C. piscicola associated with the digestive tract. C. piscicola-like isolates colonizing the various intestinal regions (foregut, midgut and hindgut) were also screened for their ability to produce growth inhibitory compounds active against the fish pathogen Aeromonas salmonicida. Of the 199 C. piscicola isolates tested, 139 inhibited growth of the pathogen.     

Microbial Ecology of the Gut in Laboratory Stocks of the Migratory Grasshopper, Melanoplus sanguinipes (Fab.) (Orthoptera: Acrididae)

Mean pH values in pooled samples of foregut, midgut, and hindgut from adult Melanoplus sanguinipes, which had been raised in the laboratory on barley shoots and wheat bran, were 5.15, 6.39, and 5.98, respectively. Homogenates of midgut/hindgut sections and frass (feces) yielded colony counts of bacteria by the spread plate method of 5.7 to 5.9 and 5.3 to 5.5 log₁₀ colonies per mg, respectively; there were no significant differences (P > 0.05) between counts obtained on several media or on media incubated aerobically or anaerobically. There was no evidence of significant populations of protozoa, fungi, or obligately anaerobic bacteria associated with the gut. A total of 168 pure strains of bacteria isolated from the gut sections were characterized and assigned to 11 taxonomic groups, including Enterococcus spp., Serratia liquefaciens, Pseudomonas spp., and Enterobacter spp. Numbers of Enterococcus spp. in the gut were 2 to 3 orders of magnitude higher than those of the other genera. Strains representing only four of the groups were recovered from bran fed to the grasshoppers; the barley shoots, which were raised in sterile soil, appeared virtually sterile. Examination of the gut wall by scanning electron microscopy revealed the presence of epimural bacteria in the foregut and hindgut but not in the midgut. The distribution of epimural cocci and bacilli differed with the gut section examined. Numerous spherical to ovoid structures up to 10 μm in diameter, which were not identified, were associated with the microvillous surface of the midgut epithelium. Acetate was present in gut, hemolymph, and frass, and it was shown that representative isolates of Enterococcus spp. and Enterobacter agglomerans produced acetate when incubated in an aqueous suspension of bran. The egestion time of solid digesta, as measured with methylene blue-stained barley shoots, was 3.0 to 5.7 h. The results show that M. sanguinipes supported extensive indigenous populations of luminal and epimural bacteria in the gut which were composed predominantly of facultatively anaerobic species; the relatively short egestion time, indicating rapid passage of digesta through the gut, was consistent with the microscopic appearance of digesta residues in frass and could account, at least in part, for the absence of a significant population of obligately anaerobic bacteria from the gut.     

The Gut Bacterial Communities Associated with Lab-Raised and Field-Collected Ants of Camponotus fragilis (Formicidae: Formicinae)

Camponotus is the second largest ant genus and known to harbor the primary endosymbiotic bacteria of the genus Blochmannia. However, little is known about the effect of diet and environment changes on the gut bacterial communities of these ants. We investigated the intestinal bacterial communities in the lab-raised and field-collected ants of Camponotus fragilis which is found in the southwestern United States and northern reaches of Mexico. We determined the difference of gut bacterial composition and distribution among the crop, midgut, and hindgut of the two types of colonies. Number of bacterial species varied with the methods of detection and the source of the ants. Lab-raised ants yielded 12 and 11 species using classical microbial culture methods and small-subunit rRNA genes (16S rRNAs) polymerase chain reaction-restriction fragment-length polymorphism analysis, respectively. Field-collected ants yielded just 4 and 1–3 species using the same methods. Most gut bacterial species from the lab-raised ants were unevenly distributed among the crop, midgut, and hindgut, and each section had its own dominant bacterial species. Acetobacter was the prominent bacteria group in crop, accounting for about 55 % of the crop clone library. Blochmannia was the dominant species in midgut, nearly reaching 90 % of the midgut clone library. Pseudomonas aeruginosa dominated the hindgut, accounting for over 98 % of the hindgut clone library. P. aeruginosa was the only species common to all three sections. A comparison between lab-raised and field-collected ants, and comparison with other species, shows that gut bacterial communities vary with local environment and diet. The bacterial species identified here were most likely commensals with little effect on their hosts or mild pathogens deleterious to colony health.     

草鱼肠道纤维素降解细菌的分离与鉴定

为更好地弄清草鱼（Ctenopharyngodon idella）肠道纤维素降解细菌的种类,采用羧甲基纤维素（CMC）作为唯一碳源的选择性培养基,分别从草鱼肠道内容物和肠道黏膜中分离到了40株产纤维素酶细菌。16S rRNA基因序列的分析结果显示,大多数产纤维素酶细菌为气单胞菌属（Aeromonas）的种类,其次为肠杆菌属（Enterobacter）的细菌以及未经分离纯培养的细菌（Uncultured bacterium）。进一步研究细菌产纤维素酶能力发现,纤维素酶活性显著性高于其他菌株的分别是A. veronii MC2、A. veronii BC6、肠杆菌科（Enterobacteriaceae）中一种未经分离纯培养的细菌BM3（Uncultured bacterium BM3）和A. jandaei HC9。草鱼肠道中简答气单胞菌（A. jandaei）、类志贺邻单胞菌（Plesiomonas shigelloides）、阴沟肠杆菌（E. cloacae）以及产气肠杆菌（E. aerogenes）是被作为产纤维素酶细菌的首次报道。       

Gut milieu shapes the bacterial communities of invasive silver carp

Silver carp is an invasive fish present in the Gobindsagar reservoir, India and has a profound impact on aquaculture. Understanding taxonomic diversity and functional attributes of gut microbiota will provide insights into the important role of bacteria in metabolism of silver carp that facilitated invasion of this exotic species. Microbial composition in foregut, midgut, hindgut and water samples was analysed using 16S rRNA gene amplicon sequencing. The bacterial communities of water samples were distinct from gut microbiota, and unique microbial assemblages were present in different regions of gut depicting profound impact of gut environment on microflora. Proteobacteria was the most abundant phyla across all samples. Ecological network analysis showed dominance of competitive interactions within posteriors region of the gut, promoting niche specialization. Predictive functional profiling revealed the microbiota specialized in digestive functions in different regions of the gut, which also reflects the dietary profile of silver carp.     

Effect of host diet on production of organic acids and methane by cockroach gut bacteria.

The effect of high-fiber diets on microbial populations and processes in cockroach guts was investigated by feeding American cockroaches (Periplaneta americana) milled cereal leaves, milled corn cob, or commercial bran-type breakfast cereal in place of the commonly used laboratory diet of dog chow. The activities and numbers of specific gut bacteria varied significantly with the insect's diet and developmental stage. Acetate and lactate were the principal organic acids present in the gut fluid of adult cockroaches and occurred at concentrations of up to 17 and 8 mM, respectively. These acids were most abundant in the gut fluid of dog chow-fed insects, and the greatest amounts were generally found in the foregut and midgut regions. Foreguts of dog chow-fed cockroaches contained an abundant population of lactic acid bacteria that formed acetate and lactate from endogenous hexoses present in the foregut. When adult cockroaches were fed dog chow amended with antibacterial drugs, (i) the concentrations of acetate, lactate, and total hexoses in gut fluid decreased significantly, (ii) the numbers of lactic acid bacteria in the foregut also decreased significantly, and (iii) the production of acetate and lactate by foregut homogenates was suppressed. It was estimated that acetate and lactate produced by bacteria in the foregut of dog chow-fed adult P. americana could support up to 14% of the insect's respiratory requirement if taken up and used by the animal. When insects were fed high-fiber diets of bran cereal, cereal leaves, or corn cob, bacterial production of acetate and lactate in the foregut diminished.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of host diet on production of organic acids and methane by cockroach gut bacteria.

The effect of high-fiber diets on microbial populations and processes in cockroach guts was investigated by feeding American cockroaches (Periplaneta americana) milled cereal leaves, milled corn cob, or commercial bran-type breakfast cereal in place of the commonly used laboratory diet of dog chow. The activities and numbers of specific gut bacteria varied significantly with the insect's diet and developmental stage. Acetate and lactate were the principal organic acids present in the gut fluid of adult cockroaches and occurred at concentrations of up to 17 and 8 mM, respectively. These acids were most abundant in the gut fluid of dog chow-fed insects, and the greatest amounts were generally found in the foregut and midgut regions. Foreguts of dog chow-fed cockroaches contained an abundant population of lactic acid bacteria that formed acetate and lactate from endogenous hexoses present in the foregut. When adult cockroaches were fed dog chow amended with antibacterial drugs, (i) the concentrations of acetate, lactate, and total hexoses in gut fluid decreased significantly, (ii) the numbers of lactic acid bacteria in the foregut also decreased significantly, and (iii) the production of acetate and lactate by foregut homogenates was suppressed. It was estimated that acetate and lactate produced by bacteria in the foregut of dog chow-fed adult P. americana could support up to 14% of the insect's respiratory requirement if taken up and used by the animal. When insects were fed high-fiber diets of bran cereal, cereal leaves, or corn cob, bacterial production of acetate and lactate in the foregut diminished.(ABSTRACT TRUNCATED AT 250 WORDS)     

松墨天牛成虫室内外种群肠道细菌的多样性及功能分析

[目的]为明确松墨天牛Monochamus alternatus成虫中、后肠细菌群落结构,探索肠道细菌的潜在功能.[方法]分别提取室外和室内饲养的松墨天牛成虫各15头(室内和室外各15个中肠、15个后肠)的肠道DNA,利用二代测序技术对松墨天牛成虫肠道细菌的16S rDNA V3-V4区序列进行测序,统计操作分类单元(...     

Enteric luminous microflora of the pond-cultured milk fishChanos chanos (Forskal)

Qualitative and quantitative investigations were made on the luminous bacteria associated with the gut of pond cultured milk fishChanos chanos. Significant differences in luminous bacterial numbers were found between gut and pond water and between gut and pond sediment, but not between pond water and sediment. No significant variation in luminous bacterial population among the gut regions was observed. The quantity of ingesta in the fish gut does not appear to influence the biomass of luminous bacteria.Vibrio harveyi andV. fischeri were the 2 most commonly encountered species, and of the 2 luminous species,V. harveyi was predominant.     

Planktonic Luminous Bacteria of Vellar Estuary,South India

Distribution of planktonic luminous bacteria in relation to environmental parameters was investigated at two stations located in the Vellar Estuary. Luminous microflora showed a pronounced seasonal cycle with very low counts during monsoon months followed by an increase in post monsoon and peak counts during summer. The population density of these procaryotes was remarkably high ranging from 3.5 to 33.1 colony forming units per ml. They constituted 2.1 to 52.1 % of the total bacterial counts. Salinity appeared to govern the distribution of luminous procaryotes as their counts corresponded well with fluctuations in salinity. Taxonomic affiliation of the isolates revealed predominance of Vibrio harveyi. Vibrio fischeri and Photobacterium leiognathi exhibited sparse distribution.     

Physicochemical conditions and microbial activities in the highly alkaline gut of the humus-feeding larva of Pachnoda ephippiata (Coleoptera: Scarabaeidae)

The soil macrofauna plays an important role in the carbon and nitrogen cycle of terrestrial ecosystems. In order to gain more insight into the role of the intestinal microbiota in transformation and mineralization of organic matter during gut passage, we characterized the physicochemical conditions, microbial activities, and community structure in the gut of our model organism, the humus-feeding larva of the cetoniid beetle Pachnoda ephippiata. Microsensor measurements revealed an extreme alkalinity in the midgut, with highest values (pH > 10) between the second and third crown of midgut ceca. Both midgut and hindgut were largely anoxic, but despite the high pH, the redox potential of the midgut content was surprisingly high even in the largest instar. However, reducing conditions prevailed in the hindgut paunch of all instars (E(h) approximately -100 mV). Both gut compartments possessed a pronounced gut microbiota, with highest numbers in the hindgut, and microbial fermentation products were present in high concentrations. The stimulation of hindgut methanogenesis by exogenous electron donors, such as H(2), formate, and methanol, together with considerable concentrations of formate in midgut and hemolymph, suggests that midgut fermentations are coupled to methanogenesis in the hindgut by an intercompartmental transfer of reducing equivalents via the hemolymph. The results of a cultivation-based enumeration of the major metabolic groups in midgut and hindgut, which yielded high titers of lactogenic, propionigenic, and acetogenic bacteria, are in good agreement not only with the accumulation of microbial fermentation products in the respective compartments but also with the results of a cultivation-independent characterization of the bacterial communities reported in the companion paper (M. Egert, B. Wagner, T. Lemke, A. Brune, and M. W. Friedrich, Appl. Environ. Microbiol. 69:6659-6668, 2003).     

Barophilic Bacteria Associated with Digestive Tracts of Abyssal Holothurians

Abyssal holothurians and sediment samples were collected at depths of 4,430 to 4,850 m in the Demerara abyssal plain. Bacterial concentrations in progressive sections of the holothurian digestive tract, as well as in surrounding surface sediments, were determined by epifluorescence microscopy. Total bacterial counts in sediments recently ingested by the animals were 1.5- to 3-fold higher than in surrounding sediments at the deepest station. Lowest counts were observed consistently in the foregut, where the digestive processes of the holothurian are believed to occur. In most animals, counts increased 3- to 10-fold in the hindgut. Microbial activity at 3°C and in situ and atmospheric pressure were determined for gut and sediment samples by measuring the utilization of [¹⁴C]glutamic acid, the doubling time of the mixed-population of culturable bacteria, and the percentage of the total bacterial count responsive to yeast extract in the presence of nalidixic acid, using epifluorescence microscopy. A barophilic microbial population, showing elevated activity under deep-sea pressure, was detected by all three methods in sediments removed from the hindgut. Transmission electron micrographs revealed intact bacteria directly associated with the intestinal lining only in the hindgut. The bacteria are believed to be carried as an actively metabolizing, commensal gut flora that transforms organic matter present in abyssal sediments ingested by the holothurian. Using data obtained in this study, it was calculated that sediment containing organic matter altered by microbial activity cleared the holothurian gut every 16 h, suggesting that abyssal holothurians and their associated gut flora are important participants in nutrient cycles of the abyssal benthic ocean.     

黑水虻消化道形态及组织结构的观察

本文比较了不同发育阶段黑水虻Hermetia illucens消化道的形态学差异,掌握了幼虫消化系统的组织学特征。利用体视镜观察黑水虻5龄幼虫、预蛹及成虫的消化道形态,利用光学显微镜和扫描电镜观察幼虫消化道各段(前肠、中肠、后肠)的显微及超微结构。结果表明：黑水虻幼虫及预蛹的消化道均由前肠(食道和前胃)、中肠及后肠组成,从幼虫到成虫,消化道的长度不断缩短。与幼虫和预蛹相比,成虫消化道形态变化明显,前胃消失,出现了嗉囊及胃盲囊,中肠进一步缩短,后肠分化为回肠、结肠和直肠。组织学观察结果显示,幼虫的唾液腺开口于口腔,由膨大的管状腺体和腺管组成。食道由特化为角质刺突的内膜层及发达的肌层组成,其末端延伸至前胃。前胃膨大为球状,包括三层组织结构。根据上皮细胞形态的差异,中肠可分为四个区段。后肠薄,肠腔内褶丰富,肠壁可见数量较多的杆状细菌。马氏管开口于中、后肠交界处,包括4支盲管,管内壁密布微绒毛。黑水虻消化道形态随发育阶段的变化,反映了各阶段摄食及消化生理的差异。幼虫消化道各段具有各自典型的组织学特征,其前、中、后肠可能分别承担了食物接纳与初步消化、消化与吸收以及重吸收功能。本研究结果为进一步了...