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Intestinal microbial flora after feeding phytohemagglutinin lectins (Phaseolus vulgaris) to rats.

Incorporation of purified phytohemagglutinin (PHA) lectins derived from red kidney beans (Phaseolus vulgaris) in the diet of weanling rats will cause growth failure, malabsorption of nutrients, and bacterial overgrowth in the small intestine. These effects are not caused by feeding a similar quantity of PHA to germfree rats. To define the morphological and bacterial changes on the mucosal surfaces of the jejunum, ileum, and cecum in greater detail, we pair fed two groups of weanling rats isocaloric, isonitrogenous diets with or without 0.5% PHA protein. On the jejunal surfaces of control rats, the mucous layer was a confluent covering with sparsely scattered bacteria and protozoa. In PHA-treated rats, the mucous layer was thin and discontinuous, and the microvillous surface of the tissue was extensively populated by bacterial cells of two distinct morphotypes--a gram-negative rod and a gram-positive coccobacillus. In all PHA-treated animals, these bacteria formed adherent monospecific or mixed adherent microcolonies on the tissue surface. Tissue damage was observed in PHA-exposed jejunal tissue as evidenced by vesiculation of the microvillous plasma membrane and by damage to the brush border membrane. On the ileal surfaces of control rats, there was a thick mucous layer within which small numbers of bacteria and protozoa were seen. Segmented filamentous bacteria were anchored in the tissue surface. In PHA-treated rats, the ileal surface was only incompletely covered by a mucous layer, and the overlying mucosal surface was extensively covered by large numbers of protozoan cells (predominantly Hexamita muris). Most of the ileal surfaces not covered by the mucous layer were occupied and virtually occluded by an overgrowth of these protozoan cells with occasional cells of Giardia muris and the tissue-associated segmented bacillus. In the ceca of control rats, the mucosa was incompletely covered by a discontinuous mucous layer and colonized by an unnamed Spirillum sp., other bacteria, and occasional protozoa. The cecal surfaces of PHA-treated rats retained most of their incomplete overlying mucous layer, which was heavily colonized by the same type of Spirillum sp. seen in untreated animals; intestinal crypts were colonized. These descriptive morphological studies demonstrate that exposure to purified PHA in the diet caused characteristic changes in the microbial ecology of the small intestine. The changes in microbial flora contributed to the malabsorption of nutrients in the small intestines of PHA-fed animals.

Animals↗

Evaluation of antimicrobial activity in Paenibacillus spp. strains isolated from natural environment.

AIMS: Paenibacillus isolates were selected to test antimicrobial activity against bacteria, filamentous fungi and yeasts isolates, with the purpose of finding new bacterium species for microbiological control. METHODS AND RESULTS: Fifty-five strains belonging to 15 species of Paenibacillus were inoculated on trypticase soya agar, potato dextrose agar and sabouraud agar plates in order to evaluate their antimicrobial activity against 16 indicator bacteria, 14 filamentous fungi and six yeasts isolates, both reference and field strains. After these screening, culture supernatant of 17 isolates was prepared. Twenty-five Paenibacillus isolates presented antimicrobial activity, where seven species (Paenibacillus chibensis; P. koreensis; P. illinoiensis; P. validu; P. pabuli; P. brasilensis and P. peoriae) stood out inhibiting at least 13 of the 16 indicator bacteria. Only 14 of the 55 isolates exhibited antifungal activity. P. peoriae inhibited 13 among the 14 filamentous fungi and all yeasts indicator strains. Fourteen isolates produced culture supernatant with antimicrobial activity. CONCLUSIONS: Among the 55 isolates analysed, 25 exhibited a broad inhibition spectrum against bacteria and pathogenic fungi. P. validus, P. chibensis, P. koreensis and P. peoriae isolates proved to be the subject matter for studies on the production of antimicrobial agents. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study revealed other two species with antimicrobial activity: P. validus and P. chibensis, and it contributed to enhance Paenibacillus biocontrolling potential, proving that it exhibit a broad action spectrum.

Anti-Infective Agents↗

Development and application of 16S rRNA-targeted probes for detection of iron- and manganese-oxidizing sheathed bacteria in environmental samples.

Comparative sequence analysis of the 16S rRNA genes from several Leptothrix and Sphaerotilus strains led to the design of an oligonucleotide probe (PS-1) based on a sequence within the hypervariable region 1 specific for four Leptothrix strains and for one of the four Sphaerotilus natans strains examined. Another probe (PSP-6) was based on a sequence within the hypervariable region 2. PSP-6 was specific for one of the two evolutionary lineages previously described for Leptothrix spp. (P. L. Siering and W. C. Ghiorse, Int. J. Syst. Bacteriol. 46:173-182, 1996). Fluorescein-labeled oligonucleotide probes were synthesized, and their specificity for fluorescence in situ hybridization identification was confirmed by a laser scanning microscopy technique (W. C. Ghiorse, D. N. Miller, R. L. Sandoli, and P. L. Siering, Microsc. Res. Tech. 33:73-86, 1996) to compare whole-cell hybridizations of closely related bacteria. Probe specificity was also tested in dot blot against total RNA isolated from four Leptothrix strains, four Sphaerotilus strains, and 15 other members of the class Proteobacteria. When the probes were tested on samples from the Sapsucker Woods wetland habitat where Leptothrix spp. are thought to play a role in manganese and iron oxidation, positive signals were obtained from several sheathed filamentous bacteria including some that were morphologically similar to previously isolated strains of "Leptothrix discophora." Other unknown filamentous sheathed bacteria also gave strong positive signals. This work provides a foundation for future studies correlating the presence of members of the Leptothrix-Sphaerotilus group of sheathed bacteria with manganese and iron oxidation activity in habitats where biological iron and manganese oxidation are important environmental processes.

Bacteria↗

Porcine colonic spirochetosis: a retrospective study of eleven cases.

This retrospective study of porcine colonic spirochetosis was done in order to characterize the clinical signs, the macroscopical and microscopical changes, and the bacteriological results of cases observed in Quebec. Necropsy records of all cases of colitis were reviewed. Eleven cases with filamentous bacteria colonizing the colonic epithelium were selected. This condition was only observed in weaned piglets, and was associated with mild persistent diarrhea and growth retardation. Macroscopic changes were generally limited to the presence of soft to liquid colonic contents. Adherence of filamentous helicoidal bacteria to the apical surface of the colonic epithelium and mild diffuse infiltration of the colonic lamina propria by mononuclear cells were the main histological findings. Weakly beta-hemolytic spirochetes were isolated in 6 cases. This condition seems to be underestimated for various reasons, and it is possible that some cases diagnosed as nonspecific colitis or superficial colitis, in fact, represent later stages of porcine colonic spirochetosis.

Animals↗

[A study of the content of pigments in the light-harvesting antenna of the green bacterium from the new family Oscillochloridaceae].

The properties of the light-harvesting superantenna of the photosynthesizing bacteria from the new family of green filamentous bacteria Oscillochloridaceae were investigated by optical spectroscopy. The antenna of Oscillochloris trichoides consists of peripheral chlorosomal and membrane subantennas. A method of isolation of Osc. trichoides chlorosomal antenna was developed using the chaothropic agent sodium thiocyanate, which simultaneously acts to stabilize chlorosomal activity. An analysis of the second derivatives of the absorption spectra of isolated chlorosomes and their acetone-methanol extracts suggested that BChl c was a predominant light-harvesting pigment in Osc. trichoides chlorosomes. Besides, it was found that, in addition to the BChl c-antenna, chlorosomes contain a minor BChl a-antenna. It was shown that the membrane BChl a-subantenna is a light-harvesting complex with absorption maxima in the near infrared region at 805 and 860 nm. Analysis of the spectral data obtained suggested that the Osc. trichoides chlorosomal antenna resembles those from Chlorobiaceae species, whereas the membrane B805-860 BChl a antenna of Osc. trichoides is close to the membrane B808-866 BChl a antenna of Chloroflexaceae species.

Bacteriochlorophylls↗

Gill diseases of cultured salmonids in Ontario.

Between 1977 and 1981, the Fish Pathology Laboratory of the Ontario Veterinary College received 239 cases from trout farms of southern Ontario, 51 (21.3%) of which had diseased gills. Branchial lesions in 86.3% of these 51 cases were characterized by marked lamellar epithelial hyperplasia with epithelial hypertrophy and lamellar fusion. Filamentous bacteria were seen on the surface of the branchial filaments and lamellae in 68.6% of the cases. Our observations highlight the importance of gill diseases as a production problem of farmed salmonids in southern Ontario.

Animals↗

Microbial Ecology of the Mango Phylloplane.

Bacteria, filamentous fungi, and yeasts were enumerated on the mango phylloplane by indirect leaf impression and washing- and dilution plating. The phylloplane microbial community was qualitatively and quantitatively related to leaf age, position in the tree canopy, seasonality, and chemical spraying. Filamentous fungi and yeasts were more abundant during winter and spring, whereas bacterial population densities increased during autumn. Community density and diversity increased progressively with leaf age. The western tree aspect sustained the least diverse bacterial, filamentous fungal, and yeast communities. Chemical sprays reduced bacterial, filamentous fungal, and yeast community diversities. Gram-positive bacteria (Bacillus spp. and coryneform spp.) exceeded gram-negative bacteria. The most common fungal genera isolated were Cladosporium and Alternaria. Yeasts prevalent in the mango phylloplane were of the genera Aureobasidium, Cryptococcus, and Sporobolomyces.

Journal Article↗

Ecological differentiation within a cosmopolitan group of planktonic freshwater bacteria (SOL cluster, Saprospiraceae, Bacteroidetes).

Members of the monophyletic SOL cluster are large filamentous bacteria inhabiting the pelagic zone of many freshwater habitats. The abundances of SOL bacteria and compositions of SOL communities in samples from 115 freshwater ecosystems around the globe were determined by fluorescence in situ hybridization with cluster- and subcluster-specific oligonucleotide probes. The vast majority (73%) of sampled ecosystems harbored SOL bacteria, and all three previously described SOL subclusters (LD2, HAL, and GKS2-217) were detected. The morphometric and chemicophysical parameters and trophic statuses of ecosystems were related to the occurrence and subcluster-specific composition of SOL bacteria by multivariate statistical methods. SOL bacteria did not occur in acidic lakes (pH < 6), and their abundance was negatively related to high trophy and pH. The subcluster-specific variation in the compositions of SOL communities could be related to the pH, electrical conductivity, altitude, and trophic status of ecosystems. All three known SOL subclusters differed in respect to their tolerated ranges of pH and conductivity. Complete niche separation was observed between the vicarious subclusters GKS2-217 and LD2; the former occurred in soft-water lakes, whereas the latter was found in a broad range of hard-water habitats. The third subgroup (HAL) showed a wide environmental tolerance and was usually found sympatrically with the LD2 or GKS2-217 subcluster. Ecological differentiation of SOL bacteria at the subcluster level was most probably driven by differential adaptation to water chemistry. The distribution of the two vicarious taxa seems to be predominantly controlled by the geological backgrounds of the catchment areas of the habitats.

Animals↗

Differentiation and developmental pathways of uropathogenic Escherichia coli in urinary tract pathogenesis.

Uropathogenic Escherichia coli (UPEC) are capable of forming complex intracellular bacterial communities (IBC) within the superficial umbrella cells of the bladders of C3H and BALB/c mice. By using time-lapse fluorescence videomicroscopy to observe infected mouse bladder explants, we discovered that IBCs formed by uropathogenic E. coli progressed through four distinct developmental stages that differed with respect to growth rate, bacterial length, colony organization, motility, and its eventual dispersal. In the first phase, bacteria in the IBC were nonmotile, rod shaped, and grew rapidly in loosely organized colonies free in the cytoplasm of the bladder superficial umbrella cells. In the second phase, the loose collection of bacteria in the IBC matured into a slower growing, highly organized biofilm-like community consisting of coccoid bacteria that ultimately filled most of the cytoplasm. In the third phase, bacteria in the biofilm-like state in the IBC switched to a motile rod-shaped phenotype allowing detachment from the community and eventual fluxing out of the host cell. During the fourth phase, the bacteria filamented. Filamentation appeared to be in response to a Toll-like receptor 4-mediated innate defense mechanism. Bacteria that fluxed out of the superficial umbrella cells were able to reenter the IBC developmental cascade but with slower kinetics and ultimately a quiescent reservoir was established. Intracellular growth and filamentation provided an advantage to the bacteria in evading infiltrating polymorphonuclear leukocytes. This work has developed a technique to observe live infected organs and revealed a complex differentiation pathway that facilitates bacterial persistence in the urinary tract.

Animals↗

Filamentous fungi and bacteria in macaroni and spaghetti products.

Filamentous fungi, especially Aspergillus flavus-oryzae, and bacteria were present in all samples of macaroni and spaghetti products tested. No aflatoxin was detected in the three samples, all relatively high in A. flavus-oryzae, that were tested for it. Five of the nine isolates of A. flavus-oryzae from macaroni and sphaghetti grown in corn, incorporated into an otherwise balanced ration, and given to ducklings resulted in death of one to five ducklings within six days.

Aflatoxins↗

Light and electron microscopic observations of a segmented filamentous bacterium attached to the mucosa of the terminal ileum of pigs.

Segmented filamentous bacteria were seen attached to apical villous enterocytes of the terminal ileum in 15 of 2,766 live pigs submitted for necropsy over a 6-year period. Infected pigs ranged in age from 2 to 13 weeks. All pigs except 2, however, were greater than 4 weeks old and had been weaned. All infected pigs came from conventional commercial herds with intensive, all-indoor, confinement rearing management systems. The bacteria were gram negative or gram variable and were not associated with any clinical disease. Bacteria were more commonly attached to epithelium on the dome villi in the ileum. Electron microscopic examination revealed organisms composed of a cranial segment with a nipple-like appendage and several other segments making up elongated filaments of various lengths. Each filament was divided into segments by transverse septa. The nipple-like appendage served as an attachment apparatus by indenting the enterocyte surface without physically penetrating it. Occasionally, the colonizing bacterium was itself colonized by small rod-shaped bacteria that completely surrounded the filament.

Animals↗

Comparison of techniques for culturing corneal ulcers.

All consecutive cases of presumed microbial keratitis were cultured with both a platinum spatula and calcium alginate swab moistened with trypticase soy broth. Ten (77%) of the 13 culture-proven ulcers in this series showed a greater number of colonies and more confluent growth on the agar plates inoculated with a moistened swab compared with a much lower yield obtained using a platinum spatula. All culture-positive bacterial ulcers showed growth when cultured with a swab. Two ulcers that were partially treated before culturing yielded very light growth from those rows inoculated with a swab and no growth from those rows inoculated with a spatula. Two ulcers showed equal growth between the two methods. One ulcer (Actinomycetes-Streptomyces/Nocardia) had greater growth obtained with a spatula compared with a swab. This series demonstrated a significantly greater retrieval of organisms from bacterial keratitis when a calcium alginate swab moistened with trypticase soy broth was used to rub the ulcer and directly inoculate the solid agar media. The single case of filamentous bacteria suggests that a spatula may yield better organism retrieval from ulcers caused by filamentous organisms.

Bacteria↗

Filamentous "Epsilonproteobacteria" dominate microbial mats from sulfidic cave springs.

Hydrogen sulfide-rich groundwater discharges from springs into Lower Kane Cave, Wyoming, where microbial mats dominated by filamentous morphotypes are found. The full-cycle rRNA approach, including 16S rRNA gene retrieval and fluorescence in situ hybridization (FISH), was used to identify these filaments. The majority of the obtained 16S rRNA gene clones from the mats were affiliated with the "Epsilonproteobacteria" and formed two distinct clusters, designated LKC group I and LKC group II, within this class. Group I was closely related to uncultured environmental clones from petroleum-contaminated groundwater, sulfidic springs, and sulfidic caves (97 to 99% sequence similarity), while group II formed a novel clade moderately related to deep-sea hydrothermal vent symbionts (90 to 94% sequence similarity). FISH with newly designed probes for both groups specifically stained filamentous bacteria within the mats. FISH-based quantification of the two filament groups in six different microbial mat samples from Lower Kane Cave showed that LKC group II dominated five of the six mat communities. This study further expands our perceptions of the diversity and geographic distribution of "Epsilonproteobacteria" in extreme environments and demonstrates their biogeochemical importance in subterranean ecosystems.

Base Sequence↗

Elimination of murine viral pathogens from the caecal contents of mice by anaerobic preparation.

The aim of this study was to investigate methods to eliminate pathogenic viral agents while preserving the 'normalizing' properties of the gut microflora of mice. Mouse hepatitis virus A59 (MHV), Reo 3 virus and Theiler GD VII virus were added to the caecal contents of 'normal' mice and following dilution, with or without subsequent culturing, given to germ-free mice. Four weeks later antibody titres against these and other viruses were determined. MHV and Theiler CD VII virus survived dilution but were eliminated during culturing. Reo-virus survived the 10(-1) dilution-culture step. All dilutions and dilution-cultures of caecal contents resulted in 'normalization' in germ-free recipients of the relative caecal weight, percentage faecal fusiform-shaped bacteria, faecal bile acids and colonization of small intestine by segmented filamentous bacteria.

Animals↗

Biological denitrification in a sequencing batch reactor.

Biological denitrification was studied in an SBR to investigate the performance. The experimental results showed that granular sludge with high activity and good settling ability can be developed smoothly in thereactor. With the granular sludge, high nitrate loading rate (0.48 kg NO3-N dm(-3) d(-1)) and removal efficiency (more than 99%) can be achieved. Scanning electronic microscope examination showed that only rod-shape bacteria existed on and within the granules, which confirmed indirectly that filamentous bacteria do not utilize nitrate and nitrite as electron acceptors. SBR is an alternative and effective technology for biological denitrification.

Ammonia↗

Filamentous bacterial population dominance in activated sludges subject to stresses.

Evolution of a filamentous bacterial population was monitored on four wastewater treatment pilot plants subject to stresses which have consisted of oxygen deficiencies and/or loading shocks. Fluorescent in situ hybridization (FISH) was used to perform filaments identification and quantification. Results obtained on the different pilot plants have led us to conclude that independently of the nature of the stresses, a single filamentous bacteria species was involved in the increase of the sludge volume index associated to the filamentous growth. In addition, when serial stresses were used, substitutions in dominant filamentous populations occurred: if another filament began to proliferate it caused the regression of the one which was formerly dominant.

Bacteria↗

16S rRNA sequence analysis of bacteria present in foaming activated sludge.

The formation of a stable viscous foam on activated sludge aeration basin surfaces is a world wide problem. Foam is usually comprised of filamentous bacteria occurring predominantly in the mixed liquor. The lack of availability of pure cultures of most filaments and the limited amount of characterisation data available for them, means that our current understanding of their taxonomic position is very poor and their relationship to other bacteria are not known. 16S rDNA sequencing was used in an attempt to identify the bacteria in a foaming sample from the Centurion Waste Water Treatment Plant in Gauteng South Africa.

Actinobacteria↗

Causes and control of filamentous growth in aerobic granular sludge sequencing batch reactors.

Poor long-term stability of aerobic granules developed in sequencing batch reactors (SBRs) remains a limitation to widespread use of aerobic granulation in treating wastewater. Filamentous growth has been commonly reported in aerobic granular sludge SBR. This review attempts to address the instability problem of aerobic granular sludge SBR from the perspective of filamentous growth in the system. The possible causes of filamentous growth are identified, including long retention times of solids, low substrate concentration in the liquid phase, high substrate gradient within the granule, dissolved oxygen deficiency in the granule, nutrient deficiency inside granule, temperature shift and flow patterns. Because of cyclic operation of aerobic granular sludge SBR and peculiarities of aerobic granules, various stresses can be present simultaneously and can result in progressive development of filamentous growth in aerobic granular sludge SBR. Overgrowth of filamentous bacteria under stress conditions appears to be a major cause of instability of aerobic granular sludge SBR. Specific recommendations are made for controlling filamentous growth.

Bacteria, Aerobic↗