Screening for Selenomonas Noxia and Akkermansia Muciniphila from the Oral Cavity of Pediatric Patients

Introduction: The human digestive tract is composed of an immense variety of microorganisms, which have been linked to many health problems such as obesity. There are many studies that demonstrate the association of cariogenic pathogens in dental patients and overall health. Selenomonas noxia, a gram negative anaerobe that is unable to metabolize sucrose and Akkermansia muciniphila, a gram negative anaerobe known to metabolize fats are two examples of pathogens that play a role in overall health. Objectives: Few studies have investigated both Selenomonas noxia (SN) and Akkermansia muciniphila (AM) with the same patient samples. The objective of this study was to analyze and evaluate the prevalence of both these organisms in a dental school-based setting. Methods: Saliva was collected from pediatric patients using an IRB-approved protocol. DNA was isolated for PCR screening and quality tested using the nanodrop. Gel electrophoresis was used for visualization. Forty seven (n=47) patients ranging from five to fifteen years of age (average 10.3) were screened. Nearly two-thirds of patients were male (64.6%) with the vast majority identified as Hispanic (72.9%) or other minority (25.0%) Results: DNA was successfully isolated with an overall average DNA concentration of 1.43 ug/uL and overall purity (A260:A80 ratio) of 1.88. Of the forty seven patients, four patients were positive for SN. There was no presence of AM in the samples. Conclusions: Research regarding AM and SN may suggest they inhabit different niches in the microbial community. The preliminary data of this pilot study suggests that SN could be found in pediatric patients while AM is not likely prevalent. However, due to the small patient sample size and large differences observed from these samples further research and analysis would need to be conducted to validate the findings.

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Lactobacillus acidophilus LA5 improves saturated fat-induced obesity mouse model through the enhanced intestinal Akkermansia muciniphila

Scientific Reports ◽

10.1038/s41598-021-85449-2 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Thunnicha Ondee ◽

Krit Pongpirul ◽

Peerapat Visitchanakun ◽

Wilasinee Saisorn ◽

Suthicha Kanacharoen ◽

...

Keyword(s):

Liver Injury ◽

Lactobacillus Acidophilus ◽

Hepg2 Cells ◽

Saturated Fat ◽

Liver Weight ◽

Gram Negative ◽

Fecal Microbiome ◽

Akkermansia Muciniphila ◽

Anti Inflammatory ◽

Inflammatory Molecules

AbstractObesity, a major healthcare problem worldwide, induces metabolic endotoxemia through the gut translocation of lipopolysaccharides (LPS), a major cell wall component of Gram-negative bacteria, causing a chronic inflammatory state. A combination of several probiotics including Lactobacillus acidophilus 5 (LA5), a potent lactic acid-producing bacterium, has previously been shown to attenuate obesity. However, data on the correlation between a single administration of LA5 versus microbiota alteration might be helpful for the probiotic adjustment. LA5 was administered daily together with a high-fat diet (HFD) for 8 weeks in mice. Furthermore, the condition media of LA5 was also tested in a hepatocyte cell-line (HepG2 cells). Accordingly, LA5 attenuated obesity in mice as demonstrated by weight reduction, regional fat accumulation, lipidemia, liver injury (liver weight, lipid compositions, and liver enzyme), gut permeability defect, endotoxemia, and serum cytokines. Unsurprisingly, LA5 improved these parameters and acidified fecal pH leads to the attenuation of fecal dysbiosis. The fecal microbiome analysis in obese mice with or without LA5 indicated; (i) decreased Bacteroidetes (Gram-negative anaerobes that predominate in non-healthy conditions), (ii) reduced total fecal Gram-negative bacterial burdens (the sources of gut LPS), (iii) enhanced Firmicutes (Gram-positive bacteria with potential benefits) and (iv) increased Verrucomycobia, especially Akkermansia muciniphila, a bacterium with the anti-obesity property. With LA5 administration, A. muciniphila in the colon were more than 2,000 folds higher than the regular diet mice as determined by 16S rRNA. Besides, LA5 produced anti-inflammatory molecules with a similar molecular weight to LPS that reduced cytokine production in LPS-activated HepG2 cells. In conclusion, LA5 attenuated obesity through (i) gut dysbiosis attenuation, partly through the promotion of A. muciniphila (probiotics with the difficulty in preparation processes), (ii) reduced endotoxemia, and (iii) possibly decreased liver injury by producing the anti-inflammatory molecules.

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Pediatric Dental Patients’ Attitudes to Rubber Dam

Journal of Clinical Pediatric Dentistry ◽

10.17796/jcpd.38.2.k73701728rh8u182 ◽

2013 ◽

Vol 38 (2) ◽

pp. 139-141 ◽

Cited By ~ 1

Author(s):

A McKay ◽

M Farman ◽

H Rodd ◽

H Zaitoun

Keyword(s):

Local Anaesthetic ◽

Pediatric Patients ◽

Young Patients ◽

Hospital Setting ◽

Radiographic Examination ◽

Dental Patients ◽

Rubber Dam ◽

Negative Score ◽

Anaesthetic Procedure ◽

Acceptance Levels

Objectives: To explore young patients’ experiences of rubber dam (RD) and determine how personal and clinical factors may influence opinions. Study design: A self-completed questionnaire was developed to capture pediatric patients’ experiences of treatment under RD in a hospital setting. Patients’ acceptance of RD and perceptions of how well it was explained to them were recorded on a 10cm Visual Analogue Scale (VAS), where zero represented the most negative score. The following clinical variables were also recorded: type of RD; procedure undertaken; use of local anaesthetic and procedure duration. Results: One hundred children (52 male, 48 female) with a mean age of 11.8 years (SD=2.29; range 7-17 years) participated. Overall, acceptance of RD was satisfactory (mean VAS=5.0). Patients were happy with the explanation of why RD was used (mean VAS=7.7). The type of RD, use of local anaesthetic, procedure undertaken and duration of the procedure did not significantly influence acceptance levels. However, RD was significantly less acceptable to patients who underwent radiographic examination whilst wearing the RD (P< 0.05, t-test). Nearly five times as many patients expressed concern at being seen wearing RD when taken to the radiography department (39.2%, n=20/51), compared to those who were reportedly self-conscious about RD when treated only on the paediatric dentistry clinic (8.2%, n=4/49). Conclusions: The use of RD appears acceptable physically and psychologically to most pediatric patients, however, visibility of the RD to others was a potential concern to some children.

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Behavior Management of Pediatric Dental Patients

PEDIATRICS ◽

10.1542/peds.90.4.651b ◽

1992 ◽

Vol 90 (4) ◽

pp. 651-652

Author(s):

◽

Richard D. Krugman ◽

Jan Bays ◽

David L. Chadwick ◽

Carolyn J. Levitt ◽

...

Keyword(s):

Behavior Management ◽

Pediatric Patients ◽

Case Reports ◽

The Other ◽

Behavioral Management ◽

Dental Patients ◽

Bladder Control ◽

Abuse And Neglect ◽

Pediatric Dentists ◽

Special Circumstances

To the Editor.— It has been brought to the attention of The American Academy of Pediatric Committee on Child Abuse and Neglect that some dentists continue to use the hand-over-mouth technique with airway restriction (HOMAR) for behavioral management of pediatric patients. We have received two case reports of children who were adversely affected by this technique. One was left with bruises and petechiae and the other lost consciousness and bladder control. The original literature on the hand over mouth (HOM) technique is alarming as it indicates that "once in a great while a patient's nostrils can be closed until he really needs air, as indicated by his color" and that 19 of 35 pediatric dentists surveyed in 1969 "felt it is permissible to completely close off a child's airway in special circumstances."

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519. Multidrug-resistant Gram-Negative Bacteremia in Pediatric Patients: Is It Time to Change to Empiric Meropenem?

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz360.588 ◽

2019 ◽

Vol 6 (Supplement_2) ◽

pp. S250-S250

Author(s):

Kanokporn Mongkolrattanothai ◽

Leslie Stach ◽

Regina Orbach

Keyword(s):

Antimicrobial Agents ◽

Pediatric Patients ◽

Multidrug Resistant ◽

Suspected Sepsis ◽

In Vitro Susceptibility ◽

Gram Negative ◽

Gram Negative Bacteremia ◽

Delayed Initiation ◽

Poor Outcomes

Abstract Background The rise of antimicrobial resistance among gram-negative (GN) pathogens has been dramatic nationally. Delayed initiation of active antimicrobial agents has been associated with poor outcomes. We aimed at evaluating the prevalence and treatment of multi-drug-resistant gram-negative (MDR-GN) bacteremia in our pediatric patients. Methods All episodes of GN bacteremia from 2017–2018 at our institution were retrospectively reviewed. GN defined as MDR in our study were carbapenem-resistant organisms (CRO), extended-spectrum β-lactamase (ESBL) producers, and GN that were resistant to cefepime and ≥2 classes of non-cephalosporin antimicrobial agents. Stenotrophomonas maltophilia was excluded. Ineffective empirical treatment (IET) is defined as an initial antibiotic regimen that is not active against the identified pathogen[s] based on in vitro susceptibility testing results. Results A total of 292 episodes of GN bacteremia were identified and 6 S. maltophilia were excluded. Of these, 29 bacteremic episodes in 26 patients were caused by MDR-GN organisms including 18 ESBL, 7 CRO, 1 ESBL and CRO, 3 non-ESBL/non-CRO cefepime-resistant MDR-GN. None of the CRO had carbapenemase genes detected. However, there was a patient with multiple sites of infection simultaneously with non-NDM CR Acinetobacter bacteremia and NDM-mediated CR-Klebsiella ventriculitis. The annual rate of MDR-GN bacteremia increased from 8% in 2017 to 12% in 2018. Almost half (48%) of episodes were community onset. Among these, all but one had underlying medical conditions with hospital exposure and most patients had central venous devices at the time of infection. 52% (15/29) episodes of MDR-GN bacteremia had IET. Despite IET, 47% (7/15) had negative blood cultures prior to initiation of effective therapy (6 ESBL and 1 P. aeruginosa). Various antibiotic regimens were used for CRO therapy as shown in Table 1. Conclusion In our institution, MDR-GN infection is increasing. As such, empiric meropenem is currently recommended in BMT or neutropenic patients with suspected sepsis. However, empiric meropenem must be used judiciously as its widely use will lead to more selection of MDR pathogens. It is essential to continue monitoring of these MDR-GN to guide appropriate empiric regimens. Disclosures All authors: No reported disclosures.

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Pseudomonas aeruginosa Coharboring BlaKPC-2 and BlaVIM-2 Carbapenemase Genes

Antibiotics ◽

10.3390/antibiotics8030098 ◽

2019 ◽

Vol 8 (3) ◽

pp. 98 ◽

Cited By ~ 1

Author(s):

Tatiana Pacheco ◽

Rosa Helena Bustos-Cruz ◽

Deisy Abril ◽

Sara Arias ◽

Lina Uribe ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Gel Electrophoresis ◽

Genetic Relationships ◽

Treatment Options ◽

Therapy Resistance ◽

Pulsed Field Gel Electrophoresis ◽

High Complexity ◽

Intrinsic Resistance ◽

Pcr Screening ◽

Extreme Drug Resistance

Pseudomonas aeruginosa, a bacterium commonly isolated from hospital settings, exhibits intrinsic resistance to a number of antibiotics and can acquire resistance during antibiotic therapy. Resistance towards carbapenems is increasing due to its overuse in the treatment of infections caused by extended-spectrum β-lactamase (ESBL) producing organisms. Nonetheless, carbapenems are essential for the treatment of high-risk infections and are one of the remaining weapons in the fight against “extreme drug resistance” of Gram-negative/positive bacilli. Herein, we describe a case report of infections caused by P. aeruginosa strains that carry blaVIM-2 and blaKPC-2 carbapenemase genes simultaneously, identified in five patients who were admitted to a high complexity health institution in Colombia. Molecular characterization included PCR screening for blaKPC, blaGES, blaOXA-48, blaIMP, blaNDM, and blaVIM carbapenemase and other resistance genes as well as analysis of the genetic relationships by genome macro-restriction and Pulsed-Field Gel Electrophoresis (PFGE) separation. In conclusion, these infections represent a major challenge to public health due to the risk of the infection spreading compounded by the fact that limited treatment options are available, thereby increasing the risk of increased morbidity and mortality.

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Polyacrylamide gel electrophoresis of exoenzymes produced by Pseudomonas fluorescens strain W

Canadian Journal of Microbiology ◽

10.1139/m71-041 ◽

1971 ◽

Vol 17 (2) ◽

pp. 241-248 ◽

Cited By ~ 2

Author(s):

Harvey Winters ◽

W. A. Corpe

Keyword(s):

Pseudomonas Fluorescens ◽

Bacterial Cell ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Deae Cellulose ◽

Complex Mixture ◽

Gram Negative ◽

Coomassie Blue ◽

The Individual ◽

Cell Envelopes

Culture filtrates from strain W of Pseudomonas fluorescens that cause lysis of gram-negative bacterial cell envelopes were examined for specific hydrolases. The enzymes were concentrated by ammonium sulfate fractionation and by column fractionation on Sephadex G-100. Attempts to separate the individual hydrolases quantitatively by elution from DEAE-cellulose failed because of the formation of aggregates. Resolution of the individual hydrolases was accomplished by disc gel electrophoresis using polyacrylamide (7.5%) gels buffered at pH 8.9 with Tris-glycine. The enzyme mixture was separated into 13 distinct protein bands which were stained with Coomassie blue. The individual hydrolases were detected either directly on the gels or by assay after elution from gel segments and included four proteinases, three phosphatases, two β-glucosidases, one ribonuclease, one lipase, one esterase, and one catalase. These methods provide a rapid, sensitive technique for the detection of many individual hydrolases in a complex mixture.

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High Incidence of Cefoxitin and Clindamycin Resistance among Anaerobes in Taiwan

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.46.9.2908-2913.2002 ◽

2002 ◽

Vol 46 (9) ◽

pp. 2908-2913 ◽

Cited By ~ 50

Author(s):

Lee-Jene Teng ◽

Po-Ren Hsueh ◽

Jui-Chang Tsai ◽

Shwu-Jen Liaw ◽

Shen-Wu Ho ◽

...

Keyword(s):

Gel Electrophoresis ◽

Antimicrobial Agents ◽

Genetic Relationships ◽

Anaerobic Bacteria ◽

Pulsed Field Gel Electrophoresis ◽

Gram Negative ◽

Pulsed Field ◽

High Incidence ◽

Single Clone ◽

Resistance Rates

ABSTRACT Susceptibilities to 16 antimicrobial agents were determined by measurement of MICs for 344 isolates of anaerobic bacteria recovered from patients with significant infections. Resistance rates varied among antimicrobial agents and the species tested. The β-lactams were more active in gram-positive than in gram-negative anaerobes. Resistance to meropenem was low (<1%). For β-lactam-β-lactamase inhibitors, piperacillin-tazobactam was most active for all species (resistance, <6%). The rates of resistance to cefoxitin (31 to 65%) and clindamycin (50 to 70%) for non-Bacteroides fragilis species of the B. fragilis group were higher than those for B. fragilis (4% resistant to cefoxitin and 33% resistant to clindamycin). Among members of B. fragilis group, Bacteroides thetaiotaomicron was the most resistant to clindamycin (70%) and cefoxitin (65%). Rates of susceptibility to imipenem and metronidazole for B. fragilis continue to be high compared to those from a previous study 10 years ago. However, resistance to metronidazole was found recently in five strains of B. fragilis. We analyzed the genetic relationships among the metronidazole-resistant B. fragilis strains by pulsed-field gel electrophoresis. The metronidazole-resistant B. fragilis strains showed genotypic heterogeneity, excluding the dissemination of a single clone.

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Viable but nonculturable coccoid forms of Prolinoborus fasciculus (Aquaspirillum fasciculus)

Canadian Journal of Microbiology ◽

10.1139/w98-071 ◽

1998 ◽

Vol 44 (9) ◽

pp. 910-912 ◽

Cited By ~ 5

Author(s):

David J Koechlein ◽

Noel R Krieg

Keyword(s):

Gel Electrophoresis ◽

Agarose Gel ◽

Agarose Gel Electrophoresis ◽

Active Growth ◽

Viable But Nonculturable ◽

Gram Negative ◽

Coccoid Form ◽

Mass Conversion ◽

Beta Hydroxybutyrate ◽

Rrna Degradation

Following active growth, the aquatic gram-negative rod Prolinoborus fasciculus (Aquaspirillum fasciculus) exhibited a mass conversion from its culturable rod form to a nonculturable coccoid form. Chloramphenicol did not prevent the conversion. Attempts to obtain variants that would not convert to the coccoid form were unsuccessful. Although the coccoid form fluoresced with acridine orange, agarose gel electrophoresis revealed extensive rRNA degradation. Poly-beta-hydroxybutyrate, abundant in the vegetative rods, was not detectable in the coccoid cells. The results suggest that the coccoid form of P. fasciculus is a degenerative form rather than part of a life cycle.Key words: Aquasprillum fasciculus, viable but nonculturable.

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Class 1 Integrons Increase Trimethoprim-Sulfamethoxazole MICs against Epidemiologically Unrelated Stenotrophomonas maltophilia Isolates

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.48.2.666-669.2004 ◽

2004 ◽

Vol 48 (2) ◽

pp. 666-669 ◽

Cited By ~ 58

Author(s):

Raquel Barbolla ◽

Mariana Catalano ◽

Betina E. Orman ◽

Angela Famiglietti ◽

Carlos Vay ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Nosocomial Infection ◽

Gel Electrophoresis ◽

Stenotrophomonas Maltophilia ◽

Pulsed Field Gel Electrophoresis ◽

Gram Negative ◽

Pulsed Field ◽

Class 1 Integrons ◽

Resistance Determinants ◽

Class 1

ABSTRACT Twenty-five plasmid-specified antimicrobial resistance determinants common to gram-negative bacilli from nosocomial infection were investigated from 31 Stenotrophomonas maltophilia isolates. Twenty-four clones were identified by pulsed-field gel electrophoresis, and in three clones that exhibited an increased trimethoprim-sulfamethoxazole MIC, the sul1 determinant was found. These results support not only the higher spread of class 1 integrons compared to other mechanisms but also the potential limitation of using trimethoprim-sulfamethoxazole for therapy of severe S. maltophilia infections.

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Some relationships between R-factor and chromosomal β-lactamase in Gram-negative bacteria

Biochemical Journal ◽

10.1042/bj1230507 ◽

1971 ◽

Vol 123 (4) ◽

pp. 507-512 ◽

Cited By ~ 10

Author(s):

J. W. Dale ◽

J. T. Smith

Keyword(s):

Gel Electrophoresis ◽

Klebsiella Aerogenes ◽

Gram Negative Bacteria ◽

Starch Gel Electrophoresis ◽

Gram Negative ◽

E Coli ◽

Molecular Weights ◽

R Factor ◽

Starch Gel ◽

R Factors

1. The β-lactamases specified by Klebsiella aerogenes 418 and the R-factor R-7268 have been partially purified. 2. The molecular weights of the K. aerogenes strains 418 and 373, Aerobacter cloacae 53, R-7268 and R-TEM β-lactamases were all about 20000; that of the enzymes from Escherichia coli strains 419 and 214T was about 31000. 3. These enzymes were also compared by means of their Km values for benzylpenicillin and ampicillin, and their behaviour on starch-gel electrophoresis. 4. The β-lactamases specified by the two Klebsiella strains, the Aerobacter strain, and the R-factors R-TEM and R-7268 were found to comprise a broadly similar group. However, within this group, only two enzymes seemed to be identical, namely those specified by the two R-factors. The two E. coli strains produce identical β-lactamases which are very different from the ‘Klebsiella/Aerobacter-type’ enzymes.

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