Anaerobic bacteria as indicators of faecal pollution

Author(s):  
D. W. F. Wheater ◽  
D. Mara ◽  
A. Opara ◽  
P. Singleton

SynopsisIn several well-authenticated instances intestinal pathogens, includingSalmonellaspecies, have been isolated from water in the absence of bacteria, such asEscherichia coli,commonly used to detect faecal pollution. The present study examines certain anaerobic, non-sporing commensals of the intestinal tract as alternative ‘indicator’ bacteria. The numbers of bifidobacteria andBacteroides fragilisrecovered from faecal specimens were between 109 and 1010 per gram while the Gram-positive, anaerobic cocci numbered only about 106per gram. In sewage, this numerical difference was eliminated by a rapid loss of viability of bifidobacteria andB. fragilisso that the counts of all three types of bacteria approximated to those ofE. coli.Storage tests with aerated and non-aerated sewage established that further loss of viability of bifidobacteria andB. fragilisoccurred only fairly slowly. In samples of water from the Dighty Water and River Tay Estuary, shown to be faecally polluted, the three anaerobic types of bacteria were recovered in numbers roughly equal to those ofE. coliand their persistence in the surface of the bed of the Dighty Water was also at least equal to that ofE. coli.These results demonstrated that, in spite of their strictly anaerobic growth requirements, Bifidobacteria,B. fragilisand the Gram-positive, anaerobic cocci persist in aerobic, aqueous environments. If their habitat can be shown to be reasonably restricted to the intestinal tract they are likely, under special circumstances, to be useful indicators of faecal pollution.

2011 ◽  
Vol 77 (14) ◽  
pp. 4894-4904 ◽  
Author(s):  
Cong T. Trinh ◽  
Johnny Li ◽  
Harvey W. Blanch ◽  
Douglas S. Clark

ABSTRACTFermentation enables the production of reduced metabolites, such as the biofuels ethanol and butanol, from fermentable sugars. This work demonstrates a general approach for designing and constructing a production host that uses a heterologous pathway as an obligately fermentative pathway to produce reduced metabolites, specifically, the biofuel isobutanol. Elementary mode analysis was applied to design anEscherichia colistrain optimized for isobutanol production under strictly anaerobic conditions. The central metabolism ofE. coliwas decomposed into 38,219 functional, unique, and elementary modes (EMs). The model predictions revealed that during anaerobic growthE. colicannot produce isobutanol as the sole fermentative product. By deleting 7 chromosomal genes, the total 38,219 EMs were constrained to 12 EMs, 6 of which can produce high yields of isobutanol in a range from 0.29 to 0.41 g isobutanol/g glucose under anaerobic conditions. The remaining 6 EMs rely primarily on the pyruvate dehydrogenase enzyme complex (PDHC) and are typically inhibited under anaerobic conditions. The redesignedE. colistrain was constrained to employ the anaerobic isobutanol pathways through deletion of 7 chromosomal genes, addition of 2 heterologous genes, and overexpression of 5 genes. Here we present the design, construction, and characterization of an isobutanol-producingE. colistrain to illustrate the approach. The model predictions are evaluated in relation to experimental data and strategies proposed to improve anaerobic isobutanol production. We also show that the endogenous alcohol/aldehyde dehydrogenase AdhE is the key enzyme responsible for the production of isobutanol and ethanol under anaerobic conditions. The glycolytic flux can be controlled to regulate the ratio of isobutanol to ethanol production.


2017 ◽  
Vol 199 (16) ◽  
Author(s):  
Brian M. Meehan ◽  
Cristina Landeta ◽  
Dana Boyd ◽  
Jonathan Beckwith

ABSTRACT Disulfide bonds are critical to the stability and function of many bacterial proteins. In the periplasm of Escherichia coli, intramolecular disulfide bond formation is catalyzed by the two-component disulfide bond forming (DSB) system. Inactivation of the DSB pathway has been shown to lead to a number of pleotropic effects, although cells remain viable under standard laboratory conditions. However, we show here that dsb strains of E. coli reversibly filament under aerobic conditions and fail to grow anaerobically unless a strong oxidant is provided in the growth medium. These findings demonstrate that the background disulfide bond formation necessary to maintain the viability of dsb strains is oxygen dependent. LptD, a key component of the lipopolysaccharide transport system, fails to fold properly in dsb strains exposed to anaerobic conditions, suggesting that these mutants may have defects in outer membrane assembly. We also show that anaerobic growth of dsb mutants can be restored by suppressor mutations in the disulfide bond isomerization system. Overall, our results underscore the importance of proper disulfide bond formation to pathways critical to E. coli viability under conditions where oxygen is limited. IMPORTANCE While the disulfide bond formation (DSB) system of E. coli has been studied for decades and has been shown to play an important role in the proper folding of many proteins, including some associated with virulence, it was considered dispensable for growth under most laboratory conditions. This work represents the first attempt to study the effects of the DSB system under strictly anaerobic conditions, simulating the environment encountered by pathogenic E. coli strains in the human intestinal tract. By demonstrating that the DSB system is essential for growth under such conditions, this work suggests that compounds inhibiting Dsb enzymes might act not only as antivirulents but also as true antibiotics.


2000 ◽  
Vol 66 (4) ◽  
pp. 1292-1297 ◽  
Author(s):  
Doo Hyun Park ◽  
J. Gregory Zeikus

ABSTRACT Neutral red (NR) was utilized as an electron mediator in microbial fuel cells consuming glucose to study both its efficiency during electricity generation and its role in altering anaerobic growth and metabolism of Escherichia coli andActinobacillus succinogenes. A study of chemical fuel cells in which NADH, NR, and ferricyanide were the electron donor, the electronophore, and the electron acceptor, respectively, showed that electrical current produced from NADH was proportional to the concentration of NADH. Fourfold more current was produced from NADH in chemical fuel cells when NR was the electron mediator than when thionin was the electron mediator. In microbial fuel cells in which E. coli resting cells were used the amount of current produced from glucose when NR was the electron mediator (3.5 mA) was 10-fold more than the amount produced when thionin was the electron mediator (0.4 mA). The amount of electrical energy generated (expressed in joules per mole of substrate) and the amount of current produced from glucose (expressed in milliamperes) in NR-mediated microbial fuel cells containing either E. coli or A. succinogeneswere about 10- and 2-fold greater, respectively, when resting cells were used than when growing cells were used. Cell growth was inhibited substantially when these microbial fuel cells were making current, and more oxidized end products were formed under these conditions. When sewage sludge (i.e., a mixed culture of anaerobic bacteria) was used in the fuel cell, stable (for 120 h) and equivalent levels of current were obtained with glucose, as observed in the pure-culture experiments. These results suggest that NR is better than other electron mediators used in microbial fuel cells and that sludge production can be decreased while electricity is produced in fuel cells. Our results are discussed in relation to factors that may improve the relatively low electrical efficiencies (1.2 kJ/mol) obtained with microbial fuel cells.


2019 ◽  
Author(s):  
Theodoulakis Christofi ◽  
Stavria Panayidou ◽  
Irini Dieronitou ◽  
Christina Michael ◽  
Yiorgos Apidianakis

AbstractGut microbiota acts as a barrier against intestinal pathogens, but species-specific protection of the host from infection remains relatively unexplored. Taking a Koch’s postulates approach in reverse to define health-promoting microbes we find thatEscherichia colinaturally colonizes the gut of healthy mice, but it is depleted from the gut of antibiotic-treated mice, which become susceptible to intestinal colonization byPseudomonas aeruginosaand concomitant mortality. Reintroduction of fecal bacteria andE. coliestablishes a high titer ofE. coliin the host intestine and increases defence againstP. aeruginosacolonization and mortality. Moreover, diet is relevant in this process because high sugars or dietary fat favoursE. colifermentation to lactic acid andP. aeruginosagrowth inhibition. To the contrary, low sugars allowP. aeruginosato produce the oxidative agent pyocyanin that inhibitsE. coligrowth. Our results provide an explanation as to whyP. aeruginosadoesn’t commonly infect the human gut, despite being a formidable microbe in lung and wound infections.Author SummaryHere we interrogate the conundrum as to whyPseudomonas aeruginosais not a clinical problem in the intestine as opposed to other tissues.P. aeruginosainteracts with Neisseria, Streptococcus, Staphylococcus and Actinomyces species found in the human lung. These are predominantly gram-positive bacteria that induceP. aeruginosavirulence. Moreover, peptidoglycan, which is abundant in gram-positive bacteria, can directly trigger the virulence ofP. aeriginosa. We reasoned thatP. aeruginosamight be benign in the human gut due to the inhibitory action of benign gram-negative intestinal bacteria, such asEscherichia coli. Therefore, we dissected the antagonism betweenE. coliandP. aeruginosaand the effect of a conventional, a fat-, a carbohydrate-and a protein-based diet in intestinal dysbiosis. Our findings support the notion that an unbalanced diet or antibiotics induces gut dysbiosis by the elimination of commensalE. coli, in addition to lactic acid bacteria, imposing a gut environment conducive toP. aeruginosainfection. Moreover, commensalE. coliprovides an explanation as to whyP. aeruginosadoesn’t commonly infect the human gut, despite being a formidable microbe in lung and wound infections.


1989 ◽  
Vol 35 (7) ◽  
pp. 698-705 ◽  
Author(s):  
Françoise Rieu ◽  
Gérard Fonty ◽  
Philippe Gouet

Characterization and enumeration of the adherent epimural community of the rumen wall of young, conventionally reared lambs were carried out from 2 to 21 days after birth. Three hundred strains were isolated by anaerobic procedures from three sites: dorsal, ventral, and caudal sacs, and from the sloughed epithelial cells. The population of epimural bacteria was very dense from the first days of the lamb's life. This population increased slightly with age. During the first week the counts were similar in the dorsal and ventral sacs, but they were 10 to 100 times lower in the caudal sac. Total counts for anaerobic bacteria were higher than the counts for aerobic bacteria. The isolated strains were distributed into 19 groups: 11 groups included aerotolerant strains, and 8 others, strictly anaerobic strains. During the first week the facultative microflora was mainly composed of Escherichia coli and Streptococci. Later, the epimural community was more complex and included Staphyloccus, Micrococcus, and Gaffkya. The strictly anaerobic microflora was mainly composed of Clostridium, Peptostreptococcus, Veillonella, Propionibacterium, and Acidaminococcus. Some of these strains appeared to be similar to those previously isolated from the rumen fluid of young lambs; however, the genera Micrococcus, Veillonella, Gaffkya, and Acidaminococcus, and E. coli seemed to be specific of the rumen wall tissues.Key words: rumen, lamb, microflora, rumen wall.


PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e3244 ◽  
Author(s):  
Thomas J. Finn ◽  
Sonal Shewaramani ◽  
Sinead C. Leahy ◽  
Peter H. Janssen ◽  
Christina D. Moon

BackgroundMany bacteria are facultative anaerobes, and can proliferate in both anoxic and oxic environments. Under anaerobic conditions, fermentation is the primary means of energy generation in contrast to respiration. Furthermore, the rates and spectra of spontaneous mutations that arise during anaerobic growth differ to those under aerobic growth. A long-term selection experiment was undertaken to investigate the genetic changes that underpin how the facultative anaerobe,Escherichia coli, adapts to anaerobic environments.MethodsTwenty-one populations ofE. coliREL4536, an aerobically evolved 10,000th generation descendent of theE. coliB strain, REL606, were established from a clonal ancestral culture. These were serially sub-cultured for 2,000 generations in a defined minimal glucose medium in strict aerobic and strict anaerobic environments, as well as in a treatment that fluctuated between the two environments. The competitive fitness of the evolving lineages was assessed at approximately 0, 1,000 and 2,000 generations, in both the environment of selection and the alternative environment. Whole genome re-sequencing was performed on random colonies from all lineages after 2,000-generations. Mutations were identified relative to the ancestral genome, and based on the extent of parallelism, traits that were likely to have contributed towards adaptation were inferred.ResultsThere were increases in fitness relative to the ancestor among anaerobically evolved lineages when tested in the anaerobic environment, but no increases were found in the aerobic environment. For lineages that had evolved under the fluctuating regime, relative fitness increased significantly in the anaerobic environment, but did not increase in the aerobic environment. The aerobically-evolved lineages did not increase in fitness when tested in either the aerobic or anaerobic environments. The strictly anaerobic lineages adapted more rapidly to the anaerobic environment than did the fluctuating lineages. Two main strategies appeared to predominate during adaptation to the anaerobic environment: modification of energy generation pathways, and inactivation of non-essential functions. Fermentation pathways appeared to alter through selection for mutations in genes such asnadR, adhE, dcuS/R, andpflB. Mutations were frequently identified in genes for presumably dispensable functions such as toxin-antitoxin systems, prophages, virulence and amino acid transport. Adaptation of the fluctuating lineages to the anaerobic environments involved mutations affecting traits similar to those observed in the anaerobically evolved lineages.DiscussionThere appeared to be strong selective pressure for activities that conferred cell yield advantages during anaerobic growth, which include restoring activities that had previously been inactivated under long-term continuous aerobic evolution of the ancestor.


2005 ◽  
Vol 52 (1-2) ◽  
pp. 101-106 ◽  
Author(s):  
R. Villemur ◽  
R. Beaudet ◽  
M. Lanthier ◽  
A. Gauthier ◽  
A. Boyer ◽  
...  

Desulfitobacterium are Gram positive, spore-forming, strictly anaerobic bacteria, that belong to the Firmicutes, Clostridia, Clostridiales, and Peptococcaceae. Most known members of the genus Desulfitobacterium have the ability to dechlorinate several halogenated compounds by a mechanism of reductive dehalogenation and use them as electron acceptors to generate energy (halorespiration). Desulfitobacteria are therefore perfect candidates to be used in bioremediation treatments of environment polluted with halogenated compounds. Understanding the physiology and the molecular mechanisms of these bacteria will help to develop better bioremediation systems. This report summarizes works that have been done in our laboratories with D. frappieri PCP-1 on reductive dehalogenases, genes encoding these dehalogenases and their expression, and the development of lab-scale PCP-degrading reactors using this bacterium.


2020 ◽  
pp. 16-18
Author(s):  
V. M. Lykhman ◽  
O. M. Shevchenko ◽  
Ye. O. Bilodid ◽  
Igor Vladimirovich Volchenko ◽  
I. A. Kulyk ◽  
...  

Among urgent surgical diseases of abdominal cavity, an acute intestinal obstruction is the most difficult to be diagnosed and treated. Leading factor, determining the development of pathophysiological processes is considered to be the progressive manifestations of enteric insufficiency syndrome, resulting in intestinal barrier impairment, negative changes in ecology of intestinal flora, increased endotoxins. To identify the small intestine microflora in acute intestinal obstruction and determine the role of dysbiotic disorders in clinical manifestations of main pathological process, a study was conducted in 60 patients with mechanical intestinal obstruction. The small intestine has a relatively rare microflora, consisting mainly of gram−positive facultative aerobic microorganisms, streptococci, lactobacilli. The distal ileum in nearly 30−55 % of healthy people contains scanty microflora, and yet the flora of this area differs from the microbial population of the higher gastrointestinal tract due to higher concentration of gram−negative bacteria. Optional−anaerobic coliform bacilli, anaerobic bifidobacteria and fusobacteria, bacteroids, the number of which starts exceeding the one of gram−positive species, are presented in significant quantities. Distal to the ileocecal valve there are significant changes in the microflora quantitative and species composition. Obligatory anaerobic bacteria become the predominant part of microflora, exceeding the number of aerobic and facultative anaerobic bacteria. The bacterial flora in different parts of gastrointestinal tract has its own specifics and is quite constant, as a result of the interaction of many factors, regulating the bacterial population in small intestine. The most important among them are: acidity of gastric juice, normal peristaltic activity of the intestine, bacterial interactions and immune mechanisms. Disorders of the intestine motor and evacuation function with its obstruction lead to slow passage of the chyme and contamination of the upper gastrointestinal tract with new types of microbes. There is a syndrome of small intestine excessive colonization, which means an increased concentration of bacterial populations in it, similar in species composition to the colon microflora. Pathological intra−intestinal contents become a source of endogenous infection and re−infection of the patient, leads to internal digestive disorders, which is manifested by syndrome of malabsorption of proteins, carbohydrates and vitamins. Key words: acute intestinal obstruction, small intestinal microflora, conditionally pathogenic microorganisms, intestinal biocenosis.


1997 ◽  
Vol 35 (11-12) ◽  
pp. 157-163 ◽  
Author(s):  
G. J. Medema ◽  
I. A. van Asperen ◽  
A. H. Havelaar

As part of a prospective cohort study among triathletes to determine a relationship between the microbiological quality of fresh bathing water and the risk of acquiring an intestinal infection, the exposure of the triathletes to microbiological contaminants was assessed. Waters were collected at seven triathlons (swimming course 1–1.5km) held in the summer of 1993 and 1994 to have a range of water qualities. All were influenced by sewage effluents, most also by agricultural run-off. Samples were collected several weeks before the event to establish a sampling programme (1993) and during the actual exposure of the triathletes (1993 and 1994) and examined for thermotolerant coliforms alone (samples preceding the event) and for E. coli, faecal enterococci, Staphylococcus aureus, F-specific RNAphages, enteroviruses (1993 and 1994) and for thermophilic Campylobacter, Salmonella, Aeromonas, Plesiomonas shigelloides and Pseudomonas aeruginosa (1993). The samples taken in the weeks before the exposure showed significant differences in thermotolerant coliform concentration between locations, depths and times. Also during swimmer exposure, significant differences occurred in microorganism levels at the different sampling points over the swimming course. As the triathletes swam as a group, they were exposed to approximately the same water at the same time. The geometric mean concentration was used to characterise each site. In the epidemiological study, the risk of an intestinal infection correlated with the concentration of thermotolerant coliforms and E. coli but not with the other parameters. The geometric mean concentration of thermotolerant coliforms at the triathlons ranged from 11–330/100mL and 54–1,200/100mL E. coli. Ranking of the seven sites by faecal pollution level, based on the geometric mean concentration of a faecal indicator, resulted in a different ranking for each indicator. At the fresh water sites studied, only the ratio between the geometric mean density of E. coli and thermotolerant coliforms was constant. The ratio between the other parameters related to faecal pollution (faecal enterococci, F-specific RNA phages, enteroviruses) varied considerably. Water quality standards relating to faecal pollution can only be based on parameters that show a significant correlation with risk of intestinal illness.


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