scholarly journals The impact of pH on Clostridioides difficile sporulation and physiology

2019 ◽  
Author(s):  
Daniela Wetzel ◽  
Shonna M. McBride
Keyword(s):  
Phenotypic Diversity ◽  
Gastrointestinal Disease ◽  
Low Ph ◽  
Spore Morphology ◽  
Clostridioides Difficile ◽  
Alkaline Environments ◽  
Acidic Conditions ◽  
Ph Conditions ◽  
The Impact ◽  
Toxin Formation

ABSTRACTClostridioides difficile is a pathogenic bacterium that infects the human colon to cause diarrheal disease. Growth of the bacterium is known to be dependent on certain bile acids, oxygen levels and nutrient availability in the intestine, but how the environmental pH can influence C. difficile is mostly unknown. Previous studies indicated that C. difficile modulates the intestinal pH, and prospective cohort studies have found a strong association between a more alkaline fecal pH and C. difficile infection. Based on these data we hypothesized that C. difficile physiology can be affected by various pH conditions. In this study, we investigated the impact of a range of pH conditions on C. difficile to assess potential effects on growth, sporulation, motility and toxin production in the strains 630∆erm and R20291. We observed pH-dependent differences in sporulation rate, spore morphology and viability. Sporulation frequency was lowest under acidic conditions, and differences in cell morphology were apparent at low pH. In alkaline environments, C. difficile sporulation was greater for strain 630∆erm, whereas R20291 produced relatively high levels of spores in a broad range of pH conditions. Rapid changes in pH during exponential growth impacted sporulation similarly among the strains. Furthermore, we observed an increase in C. difficile motility with increases in pH, and strain-dependent differences in toxin formation under acidic conditions. The data demonstrate that pH is an important parameter that affects C. difficile physiology and may reveal relevant insights into the growth and dissemination of this pathogen.IMPORTANCEClostridioides difficile is an anaerobic bacterium that causes gastrointestinal disease. C. difficile forms dormant spores, which can survive harsh environmental conditions, allowing their spread to new hosts. In this study, we determine how intestinally relevant pH conditions impact C. difficile physiology in the two divergent strains, 630∆erm and R20291. Our data demonstrate that low pH conditions reduce C. difficile growth, sporulation, and motility. However, toxin formation and spore morphology are differentially impacted in the strains at low pH. In addition, we observed that alkaline environments reduced C. difficile growth, but increased cell motility. When pH was adjusted rapidly during growth, we observed similar impacts on both strains. This study provides new insights into the phenotypic diversity of C. difficile grown under the diverse pH conditions present in the intestinal tract, and demonstrates similarities and differences in the pH responses of different C. difficile isolates.

scholarly journals The Impact of pH on Clostridioides difficile Sporulation and Physiology

2019 ◽  
Vol 86 (4) ◽  
Author(s):  
Daniela Wetzel ◽  
Shonna M. McBride
Keyword(s):  
Phenotypic Diversity ◽  
Toxin Production ◽  
Low Ph ◽  
Spore Morphology ◽  
Content Type ◽  
Clostridioides Difficile ◽  
Alkaline Environments ◽  
Acidic Conditions ◽  
Ph Conditions ◽  
The Impact

ABSTRACT Clostridioides difficile is a pathogenic bacterium that infects the human colon to cause diarrheal disease. Growth of the bacterium is known to be dependent on certain bile acids, oxygen levels, and nutrient availability in the intestine, but how the environmental pH can influence C. difficile is mostly unknown. Previous studies indicated that C. difficile modulates the intestinal pH, and prospective cohort studies have found a strong association between a more alkaline fecal pH and C. difficile infection. Based on these data, we hypothesized that C. difficile physiology can be affected by various pH conditions. In this study, we investigated the impact of a range of pH conditions on C. difficile to assess potential effects on growth, sporulation, motility, and toxin production in the strains 630Δerm and R20291. We observed pH-dependent differences in sporulation rate, spore morphology, and viability. Sporulation frequency was lowest under acidic conditions, and differences in cell morphology were apparent at low pH. In alkaline environments, C. difficile sporulation was greater for strain 630Δerm, whereas R20291 produced relatively high levels of spores in a broad range of pH conditions. Rapid changes in pH during exponential growth impacted sporulation similarly among the strains. Furthermore, we observed an increase in C. difficile motility with increases in pH, and strain-dependent differences in toxin production under acidic conditions. The data demonstrate that pH is an important parameter that affects C. difficile physiology and may reveal relevant insights into the growth and dissemination of this pathogen. IMPORTANCE Clostridioides difficile is an anaerobic bacterium that causes gastrointestinal disease. C. difficile forms dormant spores which can survive harsh environmental conditions, allowing their spread to new hosts. In this study, we determine how intestinally relevant pH conditions impact C. difficile physiology in the two divergent strains, 630Δerm and R20291. Our data demonstrate that low pH conditions reduce C. difficile growth, sporulation, and motility. However, toxin production and spore morphology were differentially impacted in the two strains at low pH. In addition, we observed that alkaline environments reduce C. difficile growth, but increase cell motility. When pH was adjusted rapidly during growth, we observed similar impacts on both strains. This study provides new insights into the phenotypic diversity of C. difficile grown under diverse pH conditions present in the intestinal tract, and demonstrates similarities and differences in the pH responses of different C. difficile isolates.

scholarly journals Variation in brachiopod microstructure and isotope geochemistry under low pH–ocean acidification–conditions

2018 ◽  
Author(s):  
Facheng Ye ◽  
Hana Jurikova ◽  
Lucia Angiolini ◽  
Uwe Brand ◽  
Gaia Crippa ◽  
...  
Keyword(s):  
Stable Isotope ◽  
Ocean Acidification ◽  
Isotope Geochemistry ◽  
Shell Growth ◽  
Low Ph ◽  
Secondary Layer ◽  
Ph Conditions ◽  
One Year ◽  
The Impact ◽  
Δ13c And Δ18o

Abstract. Throughout the last few decades and in the near future CO2–induced ocean acidification is potentially a big threat to marine calcite-shelled animals (e.g., brachiopods, bivalves, corals and gastropods). Despite the great number of studies focusing on the effects of acidification on shell growth, metabolism, shell dissolution and shell repair, the consequences on biomineral formation remain poorly understood, and only few studies addressed contemporarily the impact of acidification on shell microstructure and geochemistry. In this study, a detailed microstructure and stable isotope geochemistry investigation was performed on nine adult brachiopod specimens of Magellania venosa (Dixon, 1789), grown in the natural environment as well as in controlled culturing experiments at different pH conditions (ranging 7.35 to 8.15 ± 0.05) over different time intervals (214 to 335 days). Details of shell microstructural features, such as thickness of the primary layer, density and size of endopunctae and morphology of the basic structural unit of the secondary layer were analysed using scanning electron microscopy (SEM). Stable isotope compositions (δ13C and δ18O) were tested from the secondary shell layer along shell ontogenetic increments in both dorsal and ventral valves. Based on our comprehensive dataset, we observed that, under low pH conditions, M. venosa produced a more organic-rich shell with higher density of and larger endopunctae, and smaller secondary layer fibres, when subjected to about one year of culturing. Also, increasingly negative δ13C and δ18O values are recorded by the shell produced during culturing and are related to the CO2–source in the culture setup. Both the microstructural changes and the stable isotope results are similar to observations on brachiopods from the fossil record and strongly support the value of brachiopods as robust archives of proxies for studying ocean acidification events in the geologic past.

scholarly journals Does the natural carcinogen ptaquiloside degrade readily in groundwater?

2021 ◽  
Vol 33 (1) ◽  
Author(s):  
Jane S. Wu ◽  
Frederik Clauson-Kaas ◽  
Dan Nybro Lindqvist ◽  
Lars Holm Rasmussen ◽  
Bjarne W. Strobel ◽  
...  
Keyword(s):  
Mathematical Model ◽  
Drinking Water ◽  
Degradation Kinetics ◽  
Low Ph ◽  
Experimental Conditions ◽  
Neutral Ph ◽  
Rate Of Hydrolysis ◽  
Acidic Conditions ◽  
Ph Conditions ◽  
Ph Range

Abstract Background Ptaquiloside (PTA) is a natural carcinogen found in bracken ferns. PTA is released from the plants via soil to surface and groundwaters from where humans can be exposed via drinking water. Primary degradation of PTA is due to hydrolysis with formation of pterosin B (PTB). Temperature and pH determine the rate of hydrolysis under pure experimental conditions. To assess the applicability of the experimental model to natural groundwaters, PTA degradation kinetics were examined in a range of natural groundwaters at environmentally relevant conditions. Results PTA was quantified by UPLC-MS/MS. Over an 80-day study period, PTA half-lives ranged from 6.5 to 47 days (natural pH; 8.0 °C). The fastest degradation was observed for the most alkaline groundwaters with pH of around 8. Rates of degradation were well predicted using an existing mathematical model for hydrolysis. However, deviations from this model were found, especially at the extremes of the examined pH-range (4.7–8.2). The degree of conversion of PTA to PTB was close to unity around neutral pH. However, at slightly acidic conditions, formation of PTB could only count for 9% of the degraded PTA, indicating formation of other products. Conclusions Degradation of PTA in groundwater is determined by pH and temperature, and PTA can prevail for months under slightly acid to neutral pH conditions. The existing laboratory-based model for PTA hydrolysis is generally applicable for groundwaters but needs further validation at high and low pH.

Fermentation: an Unreliable Seed Treatment for Bacterial Fruit Blotch of Watermelon

Plant Disease ◽  
2021 ◽  
Author(s):  
Yixin Ge ◽  
Lan Luo ◽  
Liming Xia ◽  
Xue Luo ◽  
Hongkai Bi ◽  
...  
Keyword(s):  
Seed Treatment ◽  
Fruit Juice ◽  
Low Ph ◽  
Seed Infection ◽  
Vbnc State ◽  
Bacterial Fruit Blotch ◽  
Inhibition Of Growth ◽  
Ph Conditions ◽  
Global Threat ◽  
The Impact

Acidovorax citrulli is a seed-borne pathogen that causes bacterial fruit blotch (BFB), a global threat to watermelon production. Treating watermelon seeds to eliminate A. citrulli is a critical component of BFB management and several strategies have been evaluated to mitigate the impact of the disease. In China, watermelon seed producers routinely incubate seeds in watermelon juice (fermentation) to reduce the risk of seed infection by A. citrulli and seedling transmission of BFB. However, there has been limited effort to evaluate the efficacy of fermentation in mitigating A. citrulli seed infection. The current study first showed that fermented watermelon fruit juice could inhibit A. citrulli population growth, and further demonstrated that the low pH conditions, not the dynamic of temperature, generated during the fermentation process might play a major role in inhibition of growth of A. citrulli and could induce the viable but non-culturable (VBNC) state in A. citrulli. We developed an effective method that was based on PMA-PCR to detect viable A. citrulli cells under low pH conditions or in fermented watermelon fruit juice. We also provided evidence that VBNC A. citrulli cells induced by fermented watermelon fruit juice could not be resuscitated and did not retain their virulence on watermelon seedlings. However, VBNC A. citrulli cells could be resuscitated in LB medium. Based on these observations, we conclude that fermentation in watermelon fruit juice may not be an effective seed treatment for BFB as it may increase the seed infection by A. citrulli.

scholarly journals Proton Sensitivity of Corticotropin-Releasing Hormone Receptor 1 Signaling to Proopiomelanocortin in Male Mice

Endocrinology ◽  
2018 ◽  
Vol 160 (2) ◽  
pp. 276-291 ◽  
Author(s):  
Hiraku Kameda ◽  
Masaaki Yamamoto ◽  
Yukiko Tone ◽  
Masahide Tone ◽  
Shlomo Melmed
Keyword(s):  
Ligand Binding ◽  
Low Ph ◽  
Pituitary Cells ◽  
Dependent Protein Kinase ◽  
Ectopic Acth ◽  
Primary Mouse ◽  
Dependent Protein ◽  
Acidic Conditions ◽  
Mouse Pituitary ◽  
The Impact

Abstract Because an acidic cellular microenvironment is engendered by inflammation and may determine cell differentiation, we elucidated the impact of acidic conditions on induction of proopiomelanocortin (POMC) expression. Here, we demonstrate mechanisms for proton sensitivity of CRH receptor 1 (CRHR1) signaling to POMC and ACTH production. Low pH (6.8) resulted in doubling of POMC expression and ACTH production in pituitary cell line AtT-20 and in primary mouse pituitary cells. Using CRISPR knockout, we show that CRHR1 is necessary for acid-induced POMC expression, and this induction is mediated by CRHR1 histidine residues and calmodulin-dependent protein kinase II in both pituitary corticotroph cells and in nonpituitary cell lines expressing ectopic ACTH. In contrast, CRH ligand binding affinity to CRHR1 was decreased with acidic pH, implying that proton-induced POMC expression prevails in acidic conditions independently of CRH ligand binding. The results indicate that proton-induced CRHR1 signaling regulates ACTH production in response to an acidic microenvironment.

scholarly journals Variation in brachiopod microstructure and isotope geochemistry under low-pH–ocean acidification conditions

2019 ◽  
Vol 16 (2) ◽  
pp. 617-642 ◽  
Author(s):  
Facheng Ye ◽  
Hana Jurikova ◽  
Lucia Angiolini ◽  
Uwe Brand ◽  
Gaia Crippa ◽  
...  
Keyword(s):  
Stable Isotope ◽  
Ocean Acidification ◽  
Isotope Geochemistry ◽  
Shell Growth ◽  
Low Ph ◽  
Secondary Layer ◽  
Ph Conditions ◽  
Set Up ◽  
The Impact ◽  
Δ13c And Δ18o

Abstract. In the last few decades and in the near future CO2-induced ocean acidification is potentially a big threat to marine calcite-shelled animals (e.g. brachiopods, bivalves, corals and gastropods). Despite the great number of studies focusing on the effects of acidification on shell growth, metabolism, shell dissolution and shell repair, the consequences for biomineral formation remain poorly understood. Only a few studies have addressed the impact of ocean acidification on shell microstructure and geochemistry. In this study, a detailed microstructure and stable isotope geochemistry investigation was performed on nine adult brachiopod specimens of Magellania venosa (Dixon, 1789). These were grown in the natural environment as well as in controlled culturing experiments under different pH conditions (ranging from 7.35 to 8.15±0.05) over different time intervals (214 to 335 days). Details of shell microstructural features, such as thickness of the primary layer, density and size of endopunctae and morphology of the basic structural unit of the secondary layer were analysed using scanning electron microscopy. Stable isotope compositions (δ13C and δ18O) were tested from the secondary shell layer along shell ontogenetic increments in both dorsal and ventral valves. Based on our comprehensive dataset, we observed that, under low-pH conditions, M. venosa produced a more organic-rich shell with higher density of and larger endopunctae, and smaller secondary layer fibres. Also, increasingly negative δ13C and δ18O values are recorded by the shell produced during culturing and are related to the CO2 source in the culture set-up. Both the microstructural changes and the stable isotope results are similar to observations on brachiopods from the fossil record and strongly support the value of brachiopods as robust archives of proxies for studying ocean acidification events in the geologic past.

scholarly journals Does the Natural Carcinogen Ptaquiloside Degrade Readily in Groundwater?

2020 ◽  
Author(s):  
Jane S. Wu ◽  
Frederik Clauson-Kaas ◽  
Dan Nybro Lindqvist ◽  
Lars Holm Rasmussen ◽  
Bjarne W. Strobel ◽  
...  
Keyword(s):  
Drinking Water ◽  
Rate Equation ◽  
Degradation Kinetics ◽  
Low Ph ◽  
Experimental Conditions ◽  
Neutral Ph ◽  
Rate Of Hydrolysis ◽  
Acidic Conditions ◽  
Ph Conditions ◽  
Ph Range

Abstract Background: Ptaquiloside (PTA) is a natural carcinogen found in bracken ferns. PTA is released from the plants via soil to surface and groundwaters from where humans can be exposed via drinking water. Primary degradation of PTA is due to hydrolysis with formation of pterosin B (PTB). Temperature and pH determine the rate of hydrolysis under pure experimental conditions. To assess the applicability of the experimental model to natural groundwaters, PTA degradation kinetics were examined in a range of natural groundwaters at environmentally relevant conditions.Results: PTA was quantified by UPLC-MS/MS. Over an 80-day study period, PTA half-lives ranged from 6.5 to 47 days (natural pH; 8.0oC). The fastest degradation was observed for the most alkaline groundwaters with pH of around 8. Rates of degradation was well predicted using an existing rate equation for hydrolysis. However, deviations from this model were found, especially at the extremes of the examined pH-range (4.7-8.2). The rate of conversion of PTA to PTB was close to unity around neutral pH. However, at slightly acidic conditions, formation of PTB could only count for 9% of the degraded PTA, indicating formation of other products.Conclusions: Degradation of PTA in groundwater is determined by pH and temperature, and PTA can prevail for months under slightly acid to neutral pH conditions. The existing laboratory-based model for PTA hydrolysis is generally applicable for groundwaters but needs further validation at high and low pH.

scholarly journals 216. Are Automatic Antimicrobial Stop Orders an Effective Stewardship Tool for Urinary Tract and Intra-Abdominal Infections?

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S110-S110
Author(s):  
Christina Maguire ◽  
Dusten T Rose ◽  
Theresa Jaso
Keyword(s):  
Urinary Tract ◽  
Antimicrobial Therapy ◽  
Source Control ◽  
Clostridioides Difficile ◽  
Days Of Therapy ◽  
Before And After ◽  
The Difference ◽  
Tract Infections ◽  
Length Of Therapy ◽  
The Impact

Abstract Background Automatic antimicrobial stop orders (ASOs) are a stewardship initiative used to decrease days of therapy, prevent resistance, and reduce drug costs. Limited evidence outside of the perioperative setting exists on the effects of ASOs on broad spectrum antimicrobial use, discharge prescription duration, and effects of missed doses. This study aims to evaluate the impact of an ASO policy across a health system of adult academic and community hospitals for treatment of intra-abdominal (IAI) and urinary tract infections (UTI). ASO Outcome Definitions ASO Outcomes Methods This multicenter retrospective cohort study compared patients with IAI and UTI treated before and after implementation of an ASO. Patients over the age of 18 with a diagnosis of UTI or IAI and 48 hours of intravenous (IV) antimicrobial administration were included. Patients unable to achieve IAI source control within 48 hours or those with a concomitant infection were excluded. The primary outcome was the difference in sum length of antimicrobial therapy (LOT). Secondary endpoints include length and days of antimicrobial therapy (DOT) at multiple timepoints, all cause in hospital mortality and readmission, and adverse events such as rates of Clostridioides difficile infection. Outcomes were also evaluated by type of infection, hospital site, and presence of infectious diseases (ID) pharmacist on site. Results This study included 119 patients in the pre-ASO group and 121 patients in the post-ASO group. ASO shortened sum length of therapy (LOT) (12 days vs 11 days respectively; p=0.0364) and sum DOT (15 days vs 12 days respectively; p=0.022). This finding appears to be driven by a decrease in outpatient LOT (p=0.0017) and outpatient DOT (p=0.0034). Conversely, ASO extended empiric IV LOT (p=0.005). All other secondary outcomes were not significant. Ten patients missed doses of antimicrobials due to ASO. Subgroup analyses suggested that one hospital may have influenced outcomes and reduction in LOT was observed primarily in sites without an ID pharmacist on site (p=0.018). Conclusion While implementation of ASO decreases sum length of inpatient and outpatient therapy, it may not influence inpatient length of therapy alone. Moreover, ASOs prolong use of empiric intravenous therapy. Hospitals without an ID pharmacist may benefit most from ASO protocols. Disclosures All Authors: No reported disclosures

scholarly journals Reproduction in Trypanosomatids: Past and Present

Biology ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 471
Author(s):  
Camino Gutiérrez-Corbo ◽  
Bárbara Domínguez-Asenjo ◽  
María Martínez-Valladares ◽  
Yolanda Pérez-Pertejo ◽  
Carlos García-Estrada ◽  
...  
Keyword(s):  
Low Income ◽  
Phenotypic Diversity ◽  
Natural Populations ◽  
Genetic Exchange ◽  
Health Concern ◽  
Current Debate ◽  
Naturally Occurring ◽  
Experimental Works ◽  
Genomic Recombination ◽  
The Impact

Diseases caused by trypanosomatids (Sleeping sickness, Chagas disease, and leishmaniasis) are a serious public health concern in low-income endemic countries. These diseases are produced by single-celled parasites with a diploid genome (although aneuploidy is frequent) organized in pairs of non-condensable chromosomes. To explain the way they reproduce through the analysis of natural populations, the theory of strict clonal propagation of these microorganisms was taken as a rule at the beginning of the studies, since it partially justified their genomic stability. However, numerous experimental works provide evidence of sexual reproduction, thus explaining certain naturally occurring events that link the number of meiosis per mitosis and the frequency of mating. Recent techniques have demonstrated genetic exchange between individuals of the same species under laboratory conditions, as well as the expression of meiosis specific genes. The current debate focuses on the frequency of genomic recombination events and its impact on the natural parasite population structure. This paper reviews the results and techniques used to demonstrate the existence of sex in trypanosomatids, the inheritance of kinetoplast DNA (maxi- and minicircles), the impact of genetic exchange in these parasites, and how it can contribute to the phenotypic diversity of natural populations.

scholarly journals The Impact of Changing to an Algorithm-Based Clostridioides difficile Test on the Decision to Treat Clostridioides difficile

2020 ◽  
Vol 41 (S1) ◽  
pp. s407-s407
Author(s):  
Lana Dbeibo ◽  
Joy Williams ◽  
Josh Sadowski ◽  
William Fadel ◽  
Vera Winn ◽  
...  
Keyword(s):  
Critical Care ◽  
Toxic Megacolon ◽  
Adverse Outcomes ◽  
Clinical Criteria ◽  
Testing Method ◽  
Clostridioides Difficile ◽  
Critical Care Admission ◽  
The Impact ◽  
Test Result ◽  
Pcr Test

Background: Polymerase chain reaction (PCR) testing for the diagnosis of Clostridioides difficile infection (CDI) detects the presence of the organism; a positive result therefore cannot differentiate between colonization and the pathogenic presence of the bacterium. This may result in overdiagnosis, overtreatment, and risking disruption of microbial flora, which may perpetuate the CDI cycle. Algorithm-based testing offers an advantage over PCR testing as it detects toxin, which allows differentiation between colonization and infection. Although previous studies have demonstrated the clinical utility of this testing algorithm in differentiating infection from colonization, it is unknown whether the test changes CDI treatment decisions. Our facility switched from PCR to an algorithm-based testing method for CDI in June 2018. Objective: In this study, we evaluated whether clinicians’ decisions to treat patients are impacted by a test result that implies colonization (GDH+/Tox−/PCR+ test), and we examined the impact of this decision on patient outcomes. Methods: This is a retrospective cohort study of inpatients with a positive C. diff test between June 2017 and June 2019. The primary outcome was the proportion of patients treated for CDI. We compared this outcome in 3 groups of patients: those with a positive PCR test (June 2017–June 2018), those who had a GDH+/Tox−/PCR+ or a GDH+/Tox+ test result (June 2018–June 2019). Secondary outcomes included toxic megacolon, critical care admission, and mortality in patients with GDH+/Tox−/PCR+ who were treated versus those who were untreated. Results: Of patients with a positive PCR test, 86% were treated with CDI-specific antibiotics, whereas 70.4% with GDH+/Tox+ and 29.25% with GDH+/Tox−/PCR+ result were treated (P < .0001). Mortality was not different between patients with GDH+/Tox−/PCR+ who were treated versus those who were untreated (2.7% vs 3.4%; P = .12), neither was critical care admission within 2 or 7 days of test result (2% vs 1.4%; P = .15) and (4.1% vs 5.4%, P = .39), respectively. There were no cases of toxic megacolon during the study period. Conclusions: The change to an algorithm-based C. difficile testing method had a significant impact on the clinicians’ decisions to treat patients with a positive test, as most patients with a GDH+/Tox−/PCR+ result did not receive treatment. These patients did not suffer more adverse outcomes compared to those who were treated, which has implications for testing practices. It remains to be explored whether clinicians are using clinical criteria to decide whether or not to treat patients with a positive algorithm-based test, as opposed to the more reflexive treatment of patients with a positive PCR test.Funding: NoneDisclosures: None

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