scholarly journals Direct Quantification of the Enteric BacteriumOxalobacter formigenes in Human Fecal Samples by Quantitative Competitive-Template PCR

1999 ◽  
Vol 37 (5) ◽  
pp. 1503-1509 ◽  
Author(s):  
H. Sidhu ◽  
R. P. Holmes ◽  
M. J. Allison ◽  
A. B. Peck
Keyword(s):  
Kidney Stones ◽  
Internal Standard ◽  
Enteric Bacteria ◽  
Gut Flora ◽  
Oxalobacter Formigenes ◽  
Fecal Samples ◽  
Competitive Template ◽  
Dna Template ◽  
Pcr Test ◽  
Increased Susceptibility

Homeostasis of oxalic acid appears to be regulated, in part, by the gut-associated bacterium Oxalobacter formigenes. The loss of this bacterium from the gut flora is associated with an increased susceptibility to hyperoxaluria, a condition which can lead to the formation of calcium oxalate crystalluria and kidney stones. In order to identify and quantify the presence of O. formigenes in clinical specimens, a quantitative-PCR-based assay system utilizing a competitive DNA template as an internal standard was developed. This quantitative competitive-template PCR test allows for the rapid, highly specific, and reproducible quantification of O. formigenesin fecal samples and provides a prototype for development of DNA-based quantitative assays for enteric bacteria.

scholarly journals The Effect of Oxalobacter formigenes Colonization in Patients with Calcium Oxalate Renal Stones in Comparison with Healthy People in Qom: A Case-Control Study

2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Gholam Ali Jafari ◽  
Reza Fotouhi Ardakani ◽  
Jamileh Nowroozi ◽  
Mohammad Soleiman Soltanpour ◽  
Mohsen Akhavan Sepahi
Keyword(s):  
Calcium Oxalate ◽  
Kidney Stones ◽  
Renal Stones ◽  
Urinary Stones ◽  
Oxalobacter Formigenes ◽  
Fecal Samples ◽  
Urine Oxalate ◽  
Study Population ◽  
Clinical Conditions ◽  
Control Study

Background: Urinary stones are a major problem world, and their incidence has increased significantly in recent years. Objectives: This study aimed to develop a simple and rapid molecular method based on PCR and qPCR assays to detect Oxalobacter formigenes (which causes oxalate degradation in intestines) in fecal samples of healthy volunteers and patients with calcium oxalate nephrolithiasis, and determine the amount of urinary oxalate in the two groups. Methods: This study was performed on urine and fecal samples of 73 patients with kidney stones and 52 healthy individuals. After DNA extraction, PCR and qPCR assays were performed on two gene regions of Oxalobacter formigenes, OXC, and FRC. Also, urine oxalate was measured in the study population using biochemical methods. Results: We found that the presence of Oxalobacter formigenes could reduce the risk of kidney stones and calcium oxalate stones. In fact, both FRC and OXC genes were involved in the diagnosis of Oxalobacter formigenes; however, the results based on the FRC gene showed higher efficiency. In addition, the presence or absence of stones did not affect the amount of urinary excretion of oxalate, rather it is affected by diet. Conclusions: Molecular identification of Oxalobacter formigenes by PCR and qPCR assays allows rapid, specific, and reproducible detection in fecal samples, which also allows immediate processing of these samples in clinical conditions.

scholarly journals Development of a Humanized Murine Model for the Study of Oxalobacter formigenes Intestinal Colonization

2019 ◽  
Vol 220 (11) ◽  
pp. 1848-1858 ◽  
Author(s):  
Amanda M Pebenito ◽  
Menghan Liu ◽  
Lama Nazzal ◽  
Martin J Blaser
Keyword(s):  
Kidney Stones ◽  
Human Microbiome ◽  
Epidemiological Studies ◽  
Microbial Populations ◽  
Murine Models ◽  
Oxalobacter Formigenes ◽  
Human Species ◽  
Human Gut ◽  
Germ Free ◽  
Β Diversity

Abstract Background Oxalobacter formigenes are bacteria that colonize the human gut and degrade oxalate, a component of most kidney stones. Findings of clinical and epidemiological studies suggest that O. formigenes colonization reduces the risk for kidney stones. We sought to develop murine models to allow investigating O. formigenes in the context of its native human microbiome. Methods For humanization, we transplanted pooled feces from healthy, noncolonized human donors supplemented with a human O. formigenes strain into recipient mice. We transplanted microbiota into mice that were treated with broad-spectrum antibiotics to suppress their native microbiome, were germ free, or received humanization without pretreatment or received sham gavage (controls). Results All humanized mice were stably colonized with O. formigenes through 8 weeks after gavage, whereas mice receiving sham gavage remained uncolonized (P < .001). Humanization significantly changed the murine intestinal microbial community structure (P < .001), with humanized germ-free and antibiotic-treated groups overlapping in β-diversity. Both germ-free and antibiotic-treated mice had significantly increased numbers of human species compared with sham-gavaged mice (P < .001). Conclusions Transplanting mice with human feces and O. formigenes introduced new microbial populations resembling the human microbiome, with stable O. formigenes colonization; such models can define optimal O. formigenes strains to facilitate clinical trials.

scholarly journals Enteric Microbiota–Gut–Brain Axis from the Perspective of Nuclear Receptors

2018 ◽  
Vol 19 (8) ◽  
pp. 2210 ◽  
Author(s):  
Kalina Duszka ◽  
Walter Wahli
Keyword(s):  
Gut Microbiota ◽  
Complex Systems ◽  
Nuclear Receptors ◽  
Enteric Bacteria ◽  
Full Potential ◽  
Gut Flora ◽  
Current State ◽  
The Brain

Nuclear receptors (NRs) play a key role in regulating virtually all body functions, thus maintaining a healthy operating body with all its complex systems. Recently, gut microbiota emerged as major factor contributing to the health of the whole organism. Enteric bacteria have multiple ways to influence their host and several of them involve communication with the brain. Mounting evidence of cooperation between gut flora and NRs is already available. However, the full potential of the microbiota interconnection with NRs remains to be uncovered. Herewith, we present the current state of knowledge on the multifaceted roles of NRs in the enteric microbiota–gut–brain axis.

scholarly journals Oxalobacter formigenes Colonization and Oxalate Dynamics in a Mouse Model

2015 ◽  
Vol 81 (15) ◽  
pp. 5048-5054 ◽  
Author(s):  
Xingsheng Li ◽  
Melissa L. Ellis ◽  
John Knight
Keyword(s):  
Kidney Stones ◽  
Host Bacterium ◽  
Urinary Oxalate ◽  
Oxalobacter Formigenes ◽  
Content Type ◽  
Oxalate Excretion ◽  
Urinary Oxalate Excretion ◽  
Dietary Oxalate ◽  
Different Levels ◽  
Unique Ability

ABSTRACTAnimal and human studies have provided compelling evidence that colonization of the intestine withOxalobacter formigenesreduces urinary oxalate excretion and lowers the risk of forming calcium oxalate kidney stones. The mechanism providing protection appears to be related to the unique ability ofO. formigenesto rely on oxalate as a major source of carbon and energy for growth. However, much is not known about the factors that influence colonization and host-bacterium interactions. We have colonized mice withO. formigenesOxCC13 and systematically investigated the impacts of diets with different levels of calcium and oxalate onO. formigenesintestinal densities and urinary and intestinal oxalate levels. Measurement of intestinal oxalate levels in mice colonized or not colonized withO. formigenesdemonstrated the highly efficient degradation of soluble oxalate byO. formigenesrelative to other microbiota. The ratio of calcium to oxalate in diets was important in determining colonization densities and conditions where urinary oxalate and fecal oxalate excretion were modified, and the results were consistent with those from studies we have performed with colonized and noncolonized humans. The use of low-oxalate purified diets showed that 80% of animals retainedO. formigenescolonization after a 1-week dietary oxalate deprivation. Animals not colonized withO. formigenesexcreted two times more oxalate in feces than they had ingested. This nondietary source of oxalate may play an important role in the survival ofO. formigenesduring periods of dietary oxalate deprivation. These studies suggest that the mouse will be a useful model to further characterize interactions betweenO. formigenesand the host and factors that impact colonization.

scholarly journals Genome Sequence of Oxalobacter formigenes Strain HC-1

2017 ◽  
Vol 5 (27) ◽  
Author(s):  
Marguerite Hatch ◽  
Milton J. Allison ◽  
Fahong Yu ◽  
William Farmerie
Keyword(s):  
Calcium Oxalate ◽  
Genome Sequence ◽  
Kidney Stones ◽  
Kidney Stone ◽  
Human Strain ◽  
Oxalobacter Formigenes ◽  
Human Gut ◽  
Content Type

ABSTRACT The lack of Oxalobacter formigenes colonization of the human gut has been correlated with the formation of calcium oxalate kidney stones and also with the number of recurrent kidney stone episodes. Here, we present the genome sequence of HC-1, a human strain isolated from an individual residing in Iowa, USA.

scholarly journals Occurrence of potentially zoonotic and cephalosporin resistant enteric bacteria among shelter dogs in the Central and South-Central Appalachia

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Ashutosh Verma ◽  
Kimberly Carney ◽  
Marina Taylor ◽  
Kaitlyn Amsler ◽  
Joey Morgan ◽  
...  
Keyword(s):  
Public Health ◽  
Bacterial Pathogens ◽  
Enteric Bacteria ◽  
Multidrug Resistant ◽  
Fecal Samples ◽  
Microdilution Method ◽  
South Central ◽  
Broth Microdilution Method ◽  
Central Appalachia ◽  
Shelter Dogs

Abstract Background Antimicrobial resistance and presence of zoonotic enteropathogens in shelter dogs pose a public health risk to shelter workers and potential adopters alike. In this study we investigated the prevalence of zoonotic bacterial pathogens and cephalosporin resistant (CefR) enteric bacteria in the feces of apparently healthy shelter dogs in the Cumberland Gap Region (CGR) in the US states of Kentucky, Tennessee and Virginia. Results Fecal samples of 59 dogs from 10 shelters in the CGR of Central and South-Central Appalachia were screened for the presence of Campylobacter jejuni, Clostridium perfringens, Salmonella and CefR enteric bacteria. C. jejuni, C. perfringens were detected by PCR based assays. Culture and PCR were used for Salmonella detection. Of 59 dogs, fecal samples from 14 (23.7%) and 8 (13.6%) dogs tested positive for cpa and hipO genes of C. perfringens and C. jejuni, respectively. Salmonella was not detected in any of the tested samples by PCR or culture. CefR enteric bacteria were isolated on MacConkey agar supplemented with ceftiofur followed by identification using MALDI-TOF. Fecal samples from 16 dogs (27.1%) yielded a total of 18 CefR enteric bacteria. Majority of CefR isolates (14/18, 77.8%) were E. coli followed by, one isolate each of Enterococcus hirae, Acinetobacter baumannii, Acinetobacter pittii, and Pseudomonas aeruginosa. CefR enteric bacteria were tested for resistance against 19- or 24-antibiotic panels using broth microdilution method. Seventeen (94.4%) CefR bacteria were resistant to more than one antimicrobial agent, and 14 (77.8%) displayed multidrug resistance (MDR). Conclusions This study shows that shelter dogs within the CGR not only carry zoonotic bacterial pathogens, but also shed multidrug resistant enteric bacteria in their feces that may pose public health risks.

scholarly journals ENTERIC BACTERIA IN FECAL SAMPLES OF EURASIAN GRIFFON VULTURES

2014 ◽  
Vol 6 (2) ◽  
pp. 45-53
Author(s):  
Dubravka Milanov ◽  
Dragan Fabijan ◽  
Bojana Prunić ◽  
Maja Velhner ◽  
Tamaš Petrović
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Susceptibility ◽  
Salmonella Enterica ◽  
Susceptibility Testing ◽  
Enteric Bacteria ◽  
Antimicrobial Susceptibility Testing ◽  
Salmonella Spp ◽  
Fecal Samples ◽  
E Coli

Fecal samples originating from 15 Eurasian griffon vultures were collected during June 2012 in the territory of special nature reservation Uvac and examined for presence of enteric bacteria Escherichia coli and Salmonella spp. Salmonellas were isolated from five samples (33.3%) and serologically typed as Salmonella enterica subsp. enterica ser. Veneziana. E. coli was isolated from four samples (26.6%). Antimicrobial susceptibility testing revealed resistance to one and more antibiotics only in E. coli isolates.

scholarly journals Quantification of Shiga Toxin-Converting Bacteriophages in Wastewater and in Fecal Samples by Real-Time Quantitative PCR

2010 ◽  
Vol 76 (17) ◽  
pp. 5693-5701 ◽  
Author(s):  
Lejla Imamovic ◽  
Elisenda Ballesté ◽  
Juan Jofre ◽  
Maite Muniesa
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Shiga Toxin ◽  
Enteric Bacteria ◽  
Fecal Samples ◽  
Urban Sewage ◽  
Real Time Quantitative Pcr ◽  
Gene Copies ◽  
Animal Wastewater ◽  
Wastewater Samples

ABSTRACT Shiga toxin-converting bacteriophages (Stx phages) are involved in the pathogenicity of some enteric bacteria, such as Escherichia coli O157:H7. Stx phages are released from their bacterial hosts after lytic induction and remain free in the environment. Samples were analyzed for the presence of free Stx phages by an experimental approach based on the use of real-time quantitative PCR (qPCR), which enables stx to be detected in the DNA from the viral fraction of each sample. A total of 150 samples, including urban raw sewage samples, wastewater samples with fecal contamination from cattle, pigs, and poultry, and fecal samples from humans and diverse animals, were used in this study. Stx phages were detected in 70.0% of urban sewage samples (10 to 103 gene copies [GC] per ml) and in 94.0% of animal wastewater samples of several origins (10 to 1010 GC per ml). Eighty-nine percent of cattle fecal samples were positive for Stx phages (10 to 105 GC per g of sample), as were 31.8% of other fecal samples of various origins (10 to 104 GC per g of sample). The stx 2 genes and stx 2 variants were detected in the viral fraction of some of the samples after sequencing of stx 2 fragments amplified by conventional PCR. The occurrence and abundance of Stx phages in the extraintestinal environment confirm the role of Stx phages as a reservoir of stx in the environment.

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