scholarly journals Adaptive strategies of the candidate probioticE. coliNissle in the mammalian gut

2018 ◽  
Author(s):  
Nathan Crook ◽  
Aura Ferreiro ◽  
Andrew J. Gasparrini ◽  
Mitchell Pesesky ◽  
Molly K. Gibson ◽  
...  

SummaryProbiotics are living microorganisms that are increasingly used as gastrointestinal therapeutics by virtue of their innate or engineered genetic function. Unlike abiotic therapeutics, probiotics can replicate in their intended site, subjecting their genomes and therapeutic properties to natural selection. By exposing the candidate probioticE. coliNissle (EcN) to the mouse gastrointestinal tract over several weeks, we uncovered the consequences of gut transit, inter-species competition, antibiotic pressure, and engineered genetic function on the processes under selective pressure during both within-genome and horizontal evolutionary modes. We then show the utility of EcN as a chassis for engineered function by achieving the highest reported reduction in serum phenylalanine levels in a mouse model of phenylketonuria using an engineered probiotic. Collectively, we demonstrate a generalizable pipeline which can be applied to other probiotic strains to better understand their safety and engineering potential.

Author(s):  
Malireddy S Reddy

The worldwide popularity of Dr. M.S. Reddy’s Multiple Mixed Strain Probiotic Therapy to treat or prevent the hospital acquired infections (nosocomial infections) arose a great interest in the medical community around the world (Reddy and Reddy, 2016; 2017). The following questions were raised on this subject: Does Multiple Mixed Strain Probiotics directly inhibit the pathogenic bacteria (C. diff) in the gastrointestinal tract or indirectly through modulation of the host immune system or both? To be more specific, what is the exact and/or hypothetical mechanism at molecular level behind the breakthrough discovery of Dr. M.S. Reddy’s Multiple Mixed Strain Probiotic Therapy?  To answer these questions, the specific immunomodulation regulatory functions of the individual Probiotic strains (on host) have beenresearched, investigated andoutlined in this article.  A detailed explanation(s) and hypotheses have been proposed outlining the possible cumulativedirect bacteriological and indirect immunomodulatory effects (at the molecular level) of the Multiple Mixed Strain Probiotics used in Dr. M.S. Reddy’s Multiple Mixed Strain Probiotic Therapy to successfully treat C. diff infection.  A detailed scientific and research attempts were made to correlate the Probiotic induced immune activities in relation to the reduction of the symptoms associated with the hospital acquired Clostridium difficile infection during and after the Multiple Mixed Strain Probioitc Therapy.  Results of the clinical trials, microbiological tests on feces, and the clinical blood tests significantly revealed that the reasons for the success of Dr. Reddy’s Multiple Mixed Strain Probiotic Therapy are multifold. Presumably, it is predominantly due to the immunomodulatory effect they have exerted on the host immune system along with the direct inhibition of C. diff bacteria by multiple Probiotics, due to the production of bacteriocins, lactic acid and nutritional competency.In addition, the size of the individual cells of the Probiotic strains in the Multiple Mixed Strain Probiotics and their significant effect on immunomodulation has been thoroughly discussed. Results clearly proved that if Probiotics are absent in the GI tract during C. diff infection, the chances of patient survival is zero.  This is because of the excess immune stimulation and incurable damage to the epithelial cell barrier of the gastrointestinal tract caused by C. diff bacteria.  The results also revealed, without any doubt, as of to-datethe latest discovery of Dr. M.S. Reddy’s Multiple Mixed Strain Probiotic Therapy is the best way to cure the deadly hospital acquired infections affecting millions of people around the world, with high degree of mortality.  This has been attested by several practicng medical professionals and scientists around the world (Reddy and Reddy, 2017).


2020 ◽  
Vol 41 (2) ◽  
pp. 105
Author(s):  
Javier Fernando Melo-Bolívar ◽  
Ruth Yolanda Ruiz-Pardo ◽  
Michael E Hume ◽  
Hanna E Sidjabat ◽  
Luisa Marcela Villamil-Diaz

Probiotic products are viewed as an alternative to the use of antibiotics in freshwater fishes farming. Probiotic organisms include bacteria, yeast, and filamentous fungi offering different benefits to fish including growth promotion, inhibition of pathogen colonisation, and improvement of nutrient digestion, water quality, and stress tolerance, as well as enhancement of reproduction. For these reasons, this review aims to identify the main trends in probiotic amendment in freshwater fishes. Strategies to incorporate the probiotic strains in the fish feed or pellets to allow optimal viability of the strains as they reach the fish gastrointestinal tract (GIT) are crucial in probiotic research and commercial applications for freshwater fish.


1980 ◽  
Vol 29 (3) ◽  
pp. 1073-1081
Author(s):  
Rodney D. Berg

Escherichia coli C25 maintained population levels of 10 9 to 10 10 per g of cecum and translocated to 100% of the middle mesenteric lymph nodes in gnotobiotic mice monoassociated with E. coli C25. Intragastric inoculation of these mice with the cecal contents from specific-pathogen-free mice reduced the population levels of E. coli C25 to 10 6 per g of cecum and completely inhibited translocation to the mesenteric lymph nodes. Intragastric inoculation with heat-treated, Formalintreated, or filtered cecal contents did not reduce the population levels of E. coli C25 or reduce the incidence of translocation of E. coli C25 to the mesenteric lymph nodes. Thus, viable bacteria apparently are required in the cecal contents inocula to reduce the population levels and the incidence of translocation of E. coli C25. Treatment with streptomycin plus bacitracin decreased the anaerobic bacterial levels in these gnotobiotic mice, allowing increased population levels of E. coli C25 and increased translocation to the mesenteric lymph nodes. E. coli C25 also translocated to the mesenteric lymph nodes of specific-pathogen-free mice treated with streptomycin and bacitracin before colonization with E. coli C25. The high cecal population levels of E. coli C25 in these antibiotic-decontaminated specific-pathogen-free mice apparently overwhelm any barrier to translocation exerted by the immunologically developed lamina propria of the specific-pathogen-free mice. Inoculation of gnotobiotic mice with a cecal flora also reduced the population levels of an indigenous strain of E. coli with a concomitant inhibition of translocation of the indigenous E. coli to the mesenteric lymph nodes. Thus, bacterial antagonism of the gastrointestinal population levels of certain indigenous bacteria, such as E. coli , by other members of the normal bacterial flora appears to be an important defense mechanism confining bacteria to the gastrointestinal tract.


2021 ◽  
Vol 17 (2) ◽  
pp. 189-195
Author(s):  
TR Setyawati ◽  
AH Yanti ◽  
R. Kurniatuhadi

The bacterial isolates NrLtF1, NrLtF4, NrLtF5, and NrLtG2 isolated from fecal pellets and gastrointestinal tract of nypha worms (Namalycastis rhodochorde) have cellulolytic, proteolytic activity and produce organic acids. The four isolates have the potency to be developed as probiotics in nypha worm cultivation feed. This study aims to determine the probiotics potency and identify the species of NrLtF1, NrLtF4, NrLtF5, and NrLtG2 isolate based on 16srDNA sequence. The probiotic potency was carried out by the acid tolerance assays on distilled water and 0.3% acid bile media, and the antimicrobial testing against Escherichia coli (MF exp21.12). Bacterial identification was carried out by sequencing of 16sDNA sequence based on GeneBank data. The results showed that the bacterial isolates of NrLtF1, NrLtF4, NrLtF5, and NrLtG2 were able to grow on 0.3% distilled water and acid bile media. However, only the NrLtF4 and NrLtF5 inhibited E. coli (MF exp21.12) with halo zones 30 mm and 18 mm, respectively. Blasting results of the 16srDNA sequences showed that the NrLtF1, NrLtF4, NrLtF5, and NrLtG2 were closely related to Bacillus wiedmannii, Brevibacterium sediminis, Bacillus proteolyticus, and Bacillus paramycoides. The nypha worm bacterial isolates have the potency to be developed as probiotics in nypha worm culture.


2020 ◽  
Author(s):  
Elisa T. Granato ◽  
Kevin R. Foster

SUMMARYBehaviours that reliably cause the death of an actor are typically strongly disfavoured by natural selection, and yet many bacteria undergo cell lysis to release anti-competitor toxins [1–4]. This behaviour is most easily explained if only a few cells die to release toxins and help their clonemates, but the number of cells that actually lyse during bacterial warfare is unknown. The challenge is that one cannot distinguish cells that have undergone programmed suicide from those that were simply killed by a competitor’s toxin. We developed a two-colour fluorescence reporter assay in Escherichia coli to overcome this problem. Surprisingly, this revealed conditions where nearly all cells undergo programmed lysis. Adding a DNA-damaging toxin (DNase colicin) to a focal strain causes it to engage in mass cell suicide where around 85% of cells lyse to release their own toxin. Time-lapse 3D confocal microscopy revealed that self-lysis occurs at even higher frequencies (~94%) at the interface between competing colonies. We sought to understand how such high levels of cell suicide could be favoured by natural selection. Exposing E. coli that do not perform lysis to the DNase colicin revealed that mass lysis only occurs when cells are going to die anyway from toxin exposure. From an evolutionary perspective, this renders the behaviour cost-free as these cells have zero reproductive potential. This explains how mass cell suicide can evolve, as any small benefit to surviving clonemates can lead to the strategy being favoured by natural selection. Our findings have strong parallels to the suicidal attacks of social insects [5–8], which are also performed by individuals with low reproductive potential, suggesting convergent evolution in these very different organisms.HIGHLIGHTSA novel assay can detect Escherichia coli undergoing cell suicide to release toxinsWe quantified the frequency of suicidal self-lysis during competitionsUnder some conditions, nearly all cells will self-lyse to release toxinsSelf-lysis makes evolutionary sense as cells will die anyway from competitors’ toxins


2019 ◽  
Author(s):  
Waqas Chaudhry ◽  
Nicole Vega ◽  
Adithi Govindan ◽  
Rodrigo Garcia ◽  
Esther Lee ◽  
...  

AbstractBacteriophages are deemed either lytic (virulent) or temperate, respectively depending on whether their genomes are transmitted solely horizontally, or both horizontally and vertically. To elucidate the ecological and evolutionary conditions under which natural selection will favor the evolution and maintenance of lytic or temperate modes of phage replication and transmission, we use a comprehensive mathematical model of the dynamics of temperate and virulent phage in populations of bacteria sensitive and resistant to these viruses. For our numerical analysis of the properties of this model, we use parameters estimated with the temperate bacteriophage Lambda, λ, it’s clear and virulent mutants, andE. colisensitive and resistant - refractory to these phages. Using batch and serial transfer population dynamic and reconstruction experiments, we test the hypotheses generated from this theoretical analysis. Based on the results of this jointly theoretical and experimental study, we postulate the conditions under which natural selection will favor the evolution and maintenance of lytic and temperate modes of phage replication and transmission. A compelling and novel prediction thisin silico,in vitro, andin plasticostudy makes is lysogenic bacteria from natural populations will be resistant-refractory to the phage for which they are lysogenic as well as lytic phage sharing the same receptors as these temperate viruses.


2012 ◽  
Vol 78 (19) ◽  
pp. 6799-6803 ◽  
Author(s):  
Sam Abraham ◽  
David M. Gordon ◽  
James Chin ◽  
Huub J. M. Brouwers ◽  
Peter Njuguna ◽  
...  

ABSTRACTThe role ofEscherichia colias a pathogen has been the focus of considerable study, while much less is known about it as a commensal and how it adapts to and colonizes different environmental niches within the mammalian gut. In this study, we characterizeEscherichia coliorganisms (n= 146) isolated from different regions of the intestinal tracts of eight pigs (dueodenum, ileum, colon, and feces). The isolates were typed using the method of random amplified polymorphic DNA (RAPD) and screened for the presence of bacteriocin genes and plasmid replicon types. Molecular analysis of variance using the RAPD data showed thatE. coliisolates are nonrandomly distributed among different gut regions, and that gut region accounted for 25% (P< 0.001) of the observed variation among strains. Bacteriocin screening revealed that a bacteriocin gene was detected in 45% of the isolates, with 43% carrying colicin genes and 3% carrying microcin genes. Of the bacteriocins observed (H47, E3, E1, E2, E7, Ia/Ib, and B/M), the frequency with which they were detected varied with respect to gut region for the colicins E2, E7, Ia/Ib, and B/M. The plasmid replicon typing gave rise to 25 profiles from the 13 Inc types detected. Inc F types were detected most frequently, followed by Inc HI1 and N types. Of the Inc types detected, 7 were nonrandomly distributed among isolates from the different regions of the gut. The results of this study indicate that not only may the different regions of the gastrointestinal tract harbor different strains ofE. colibut also that strains from different regions have different characteristics.


2020 ◽  
Vol 117 (17) ◽  
pp. 9519-9528 ◽  
Author(s):  
Natalie Sirisaengtaksin ◽  
Max A. Odem ◽  
Rachel E. Bosserman ◽  
Erika M. Flores ◽  
Anne Marie Krachler

Enterohemorrhagic Escherichia coli (EHEC) is a foodborne pathogen that colonizes the gastrointestinal tract and has evolved intricate mechanisms to sense and respond to the host environment. Upon the sensation of chemical and physical cues specific to the host’s intestinal environment, locus of enterocyte effacement (LEE)-encoded virulence genes are activated and promote intestinal colonization. The LEE transcriptional activator GrlA mediates EHEC’s response to mechanical cues characteristic of the intestinal niche, including adhesive force that results from bacterial adherence to epithelial cells and fluid shear that results from intestinal motility and transit. GrlA expression and release from its inhibitor GrlR was not sufficient to induce virulence gene transcription; mechanical stimuli were required for GrlA activation. The exact mechanism of GrlA activation, however, remained unknown. We isolated GrlA mutants that activate LEE transcription, independent of applied mechanical stimuli. In nonstimulated EHEC, wild-type GrlA associates with cardiolipin membrane domains via a patch of basic C-terminal residues, and this membrane sequestration is disrupted in EHEC that expresses constitutively active GrlA mutants. GrlA transitions from an inactive, membrane-associated state and relocalizes to the cytoplasm in response to mechanical stimuli, allowing GrlA to bind and activate the LEE1 promoter. GrlA expression and its relocalization in response to mechanical stimuli are required for optimal virulence regulation and colonization of the host intestinal tract during infection. These data suggest a posttranslational regulatory mechanism of the mechanosensor GrlA, whereby virulence gene expression can be rapidly fine-tuned in response to the highly dynamic spatiotemporal mechanical profile of the gastrointestinal tract.


2013 ◽  
Vol 59 (11) ◽  
pp. 720-725 ◽  
Author(s):  
E. Styková ◽  
R. Nemcová ◽  
I. Valocký ◽  
F. Novotný ◽  
P. Guba

In the present study, we examined the adherence of indigenous vaginal bacteria, probiotic strains, and metritis pathogens to mucus collected from different parts of the reproductive tracts of heifers and cows and compared their adherence with the bacterial adherence to mucus collected from the stomach and large intestine of pigs. Most of the vaginal strains adhered to mucus collected from different parts of the reproductive tract and strongly adhered to gastric mucus, with the exception of Lactobacillus buchneri 24S8. Only Lactobacillus mucosae 29S8, Enterococcus faecium E21, and E. faecium EAC adhered to colonic mucus. Probiotic strains adhered strongly to mucus collected from the reproductive tract and gastric mucus but did not adhere to colonic mucus. Pathogenic strains were adherent to vaginal, uterine horn, and gastric mucus, except Escherichia coli O8:K88ab:H9 (65), Fusobacterium necrophorum, and Gardnerella vaginalis, which adhered to uterine cervix mucus. Only Kocuria kristinae and G. vaginalis adhered to uterine body mucus; E. coli O149:K88ac (EC) adhered to colonic mucus. The strains did not exhibit host specificity but rather strain specificity. The ability to adhere to mucus was a characteristic unique to each strain. To our knowledge, this is the first report regarding in vitro adherence of GRAS (Generally Regarded As Safe) lactobacilli isolated from different sources to mucus collected from different parts of the reproductive tract.


2020 ◽  
Vol 175 ◽  
pp. 03003
Author(s):  
Yuri Kozlovsky ◽  
Tatiana Khomyakova ◽  
Aminat Magomedova ◽  
Galina Kozlovskaya ◽  
Mariya Zinevich

Gut microbiome changes can be considered as a causative factor of stress-associated intestinal diseases. The aim of the investigation was to study the effects of the oral probiotic strains Lactobacillus casei LB 148 and E. coli EB 387 onto gut microbiota, as well as the intestinal immune system of rats Sprague Dawley in a healthy state and under cold stress. There are no strict official recommendations here in the Russian Federation for preclinical studies of new probiotic strains. Here we describe a method used to estimate the safety and effectiveness of the oral use of a probiotic. It is cheap and simple, so we recommend using it at least for the screening search of effective and safe probiotics. The number of Peyer’s patches in the small intestine of rats in different compartments was counted and then compared between different groups of rats. After cold stress there was a change in the number of Peyer’s patches together with microbiota shifts. Lactobacillus casei LB 148 as well as E. coli EB 387 protected the immune system against the stress effects.


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