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Anaerobe Apr 2020In microbiological diagnosis of periprosthetic joint infection (PJI) there is much controversial discussion about culture media and incubation time, especially if...
BACKGROUND
In microbiological diagnosis of periprosthetic joint infection (PJI) there is much controversial discussion about culture media and incubation time, especially if anaerobic bacteria are the causative agents. This retrospective analysis was conducted to compare the results obtained by inoculation of sonicate fluid from prosthetic components into BD Bactec blood culture bottles with those obtained by our culture method using sensitive supplemented growth media.
METHODS
Twenty-eight cases were included in this study. For definition of PJI, the criteria of the Musculoskeletal Infection Society (MSIS) were considered. The quantity and time to positivity of anaerobes detected in sonicate fluid were monitored both from inoculated supplemented liver thioglycollate broth and anaerobic blood culture bottles. Furthermore, phenotypic testing was performed on the antimicrobial activity within the sonicate fluid.
RESULTS
The most frequently isolated microbes were Cutibacterium species, followed by Finegoldia magna, Parvimonas micra, Robinsoniella peoriensis, Clostridium species, Peptoniphilus harei and Slackia exigua. In 24 cases, the microorganisms became detectable within five days (median time 3.2 days) when sonicate fluid was incubated in supplemented liver thioglycollate broth, regardless of whether the patients had taken antimicrobial agents prior to surgery. However, when sonicate fluid was inoculated into anaerobic Bactec bottles, the median time to positivity was 7.4 days and only 12 cases (43%) were correctly identified. Sixteen cases remained negative after 14 days of incubation.
CONCLUSION
Depending on the pathogen, incubation of sonicate fluid using blood culture bottles can support diagnosis of PJI but compared with our culture medium it is less efficient if anaerobes are the suspected cause of infection. Microbiological expertise is therefore indispensable to ensure reliable detection of these microorganisms in PJI until a gold standard for laboratory handling of anaerobes has been established.
Topics: Aged; Aged, 80 and over; Bacteria, Anaerobic; Bacterial Infections; Blood Culture; Female; Humans; Male; Microbiological Techniques; Middle Aged; Prosthesis-Related Infections; Retrospective Studies; Sonication
PubMed: 32001335
DOI: 10.1016/j.anaerobe.2020.102152 -
Anaerobe Feb 2015The antibiotic susceptibility profile of the Bacteroides fragilis group, Gram-positive anaerobic cocci (GPAC), Fusobacterium spp., Prevotella spp., Veillonella spp. and...
The antibiotic susceptibility profile of the Bacteroides fragilis group, Gram-positive anaerobic cocci (GPAC), Fusobacterium spp., Prevotella spp., Veillonella spp. and Bilophila wadsworthia for amoxicillin, amoxicillin-clavulanic acid, clindamycin and metronidazole was determined. Human clinical isolates were isolated between 2011 and 2013 at the Microbiological Diagnostic Laboratory of the University Medical Center Groningen, The Netherlands and subjected to MALDI-TOF MS identification and susceptibility testing using E-test for MIC determination. Differences in clindamycin susceptibility between species of the B. fragilis group and GPAC were observed, with Bacteroides ovatus and Peptoniphilus harei having the highest resistance rates. Compared to other European countries, in The Netherlands the MIC90 for clindamycin of fusobacteria is low. Metronidazole resistance was first encountered in the genus Prevotella in 2013, but not in species of GPAC as reported in Belgium and Bulgaria. The differences in clindamycin resistance between the different European countries and reports of metronidazole resistance within the genera Prevotella and GPAC warrant more extensive susceptibility studies on anaerobic pathogens.
Topics: Anti-Bacterial Agents; Bacteria, Anaerobic; Bacterial Infections; Drug Resistance, Bacterial; Humans; Microbial Sensitivity Tests; Netherlands; Prevalence; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 25192966
DOI: 10.1016/j.anaerobe.2014.08.011 -
Anaerobe Oct 2017Secnidazole, a 5-nitroimidazole with a longer half-life, is structurally related to metronidazole and tinidazole. For treatment of bacterial vaginosis (BV), secnidazole... (Comparative Study)
Comparative Study
Secnidazole, a 5-nitroimidazole with a longer half-life, is structurally related to metronidazole and tinidazole. For treatment of bacterial vaginosis (BV), secnidazole is a suitable single-dose oral drug having a longer serum half-life than metronidazole. The objective of this study was to evaluate the antimicrobial susceptibility of vaginal isolates of facultative and anaerobic bacteria to secnidazole, metronidazole, tinidazole and clindamycin. A total of 605 unique BV-related bacteria and 108 isolates of lactobacilli recovered from the human vagina of US women during the years 2009-2015 were tested for antimicrobial susceptibility by the agar dilution CLSI reference method to determine the minimal inhibitory concentration (MIC). The MIC (μg/mL) for secnidazole was similar to metronidazole and tinidazole for Anaerococcus tetradius (secnidazole: MIC 2; metronidazole: MIC 2; tinidazole: MIC 4), Atopobium vaginae (32; >128; 128), Bacteroides species (2; 2; 2), Finegoldia magna (2; 2; 4), Gardnerella vaginalis (128; 64; 32), Mageeibacillus indolicus (2; 2; 2), Megasphaera-like bacteria (0.5; 0.25; 0.5), Mobiluncus curtisii (128; >128; >128) and Mobiluncus mulieris (>128; >128; >128), Peptoniphilus lacrimalis (4; 4; 4) and Peptoniphilus harei (2; 2; 4), Porphyromonas species (0.25; 0.5; 0.25), Prevotella bivia (8; 8; 8), Prevotella amnii (2; 1; 2) and Prevotella timonensis (2; 2; 2). In this evaluation, 14 (40%) of 35 P. bivia, 5 (14%) of 35 P. amnii and 21 (58%) of 36 P. timonensis isolates were resistant to clindamycin with MIC values of >128 μg/mL. Secnidazole, like metronidazole, was superior to clindamycin for Prevotella spp., Bacteroides spp., Peptoniphilus spp., Anaerococcus tetradius and Finegoldia magna. Clindamycin had greater activity against Atopobium vaginae, Gardnerella vaginalis and Mobiluncus spp. compared to the nitroimidazoles. All 27 Lactobacillus crispatus, 26 (96%) of 27 L. jensenii, 5 (19%) of 27 L. gasseri and 18 (67%) of 27 L. iners isolates were susceptible to clindamycin (MIC ≤2) while the MIC for all lactobacilli tested was >128 μg/mL for secnidazole, metronidazole and tinidazole. Secnidazole has similar in vitro activity against the range of microorganisms associated with BV compared to metronidazole or tinidazole. Further, secnidazole spares lactobacilli, a characteristic which is desirable in drugs used to treat bacterial vaginosis.
Topics: Anti-Bacterial Agents; Azoles; Bacteria; Clindamycin; Female; Humans; Microbial Sensitivity Tests; United States; Vaginosis, Bacterial
PubMed: 28522362
DOI: 10.1016/j.anaerobe.2017.05.005 -
Clinical Microbiology and Infection :... Oct 2007Fluorescent probes targeted at 16S rRNA were designed for Peptostreptococcus anaerobius and Peptostreptococcus stomatis (Pana134), Parvimonas micra (Pamic1435),...
Fluorescent probes targeted at 16S rRNA were designed for Peptostreptococcus anaerobius and Peptostreptococcus stomatis (Pana134), Parvimonas micra (Pamic1435), Finegoldia magna (Fmag1250), Peptoniphilus asaccharolyticus (Pnasa1254), Peptoniphilus ivorii (Pnivo731), Peptoniphilus harei (Pnhar1466), Anaerococcus vaginalis (Avag1280) and Anaerococcus lactolyticus (Alac1438), based on the 16S rRNA sequences of reference strains and 88 randomly chosen clinical isolates. These strains were also used for validation of the probes. Application of the probes to an additional group of 100 clinical isolates revealed that 87% of Gram-positive anaerobic cocci (GPAC) could be identified with this set of probes. The 16S rRNAs of 13 clinical isolates that could not be identified were sequenced. Most of these isolates were GPAC that were not targeted by the probes. No clinical isolates of Pn. asaccharolyticus were encountered. Near full-length sequences were obtained from 71 of 101 (n = 88 + 13) sequenced clinical isolates. Of these, 25 showed <98% similarity with the homologues of the closest established species. The Fmag1250, Pamic1435, Pnhar1466, Pana134, Pnasa1254 and Pnivo731 probes allowed reliable identification and hybridised with all corresponding isolates. The Avag1280 and Alac1438 probes failed to hybridise with two isolates and one isolate, respectively, because of intra-species variation. However, overall, the set of probes yielded fast and reliable identification for the majority of clinical isolates.
Topics: Anaerobiosis; Bacterial Typing Techniques; DNA, Bacterial; DNA, Ribosomal; Fluorescent Dyes; Gram-Positive Bacterial Infections; Gram-Positive Cocci; Humans; Phylogeny; RNA, Ribosomal, 16S; Reference Standards; Sensitivity and Specificity; Sequence Analysis, DNA; Species Specificity
PubMed: 17714522
DOI: 10.1111/j.1469-0691.2007.01803.x -
Antimicrobial Agents and Chemotherapy Mar 2011The susceptibility of 14 species of 115 Gram-positive anaerobic cocci (GPAC) was determined for 14 antibiotics. To assure correct identification, strains were...
The susceptibility of 14 species of 115 Gram-positive anaerobic cocci (GPAC) was determined for 14 antibiotics. To assure correct identification, strains were genotypically identified by fluorescence in situ hybridization and sequencing. Susceptibility differences (MIC₅₀ and MIC₉₀) for penicillin G, clindamycin, tigecycline, levofloxacin, amoxicillin-clavulanic acid, cefoxitin, ertapenem, meropenem, metronidazole, and doxycycline were found for the three clinically most relevant GPAC species: Finegoldia magna, Parvimonas micra, and Peptoniphilus harei.
Topics: Anti-Bacterial Agents; Bacteria, Anaerobic; Gram-Positive Cocci; Microbial Sensitivity Tests; Netherlands
PubMed: 21189338
DOI: 10.1128/AAC.01771-09 -
Journal of Pathogens 2015In total, 81 nonduplicate gram-positive anaerobic cocci (GPAC) were involved in this study. The GPAC were isolated from samples collected from cancer patients between...
In total, 81 nonduplicate gram-positive anaerobic cocci (GPAC) were involved in this study. The GPAC were isolated from samples collected from cancer patients between 2004 and 2014. Species identification was carried out by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). The majority of isolates were identified as Finegoldia magna (47%) and Peptoniphilus harei (28%). The susceptibility of six species of GPAC was determined for eight antibiotics according to E-test methodology. Furthermore, all isolates were susceptible to imipenem, vancomycin, and linezolid. Susceptibility to penicillin G, amoxicillin/clavulanate, metronidazole, ciprofloxacin, and levofloxacin varied for different species. One Finegoldia magna isolate was multidrug-resistant (i.e., parallel resistance to five antimicrobial agents, including metronidazole, was observed). Two Parvimonas micra isolates were highly resistant to metronidazole (MIC 256 μg/mL) but were sensitive to other tested antibiotics.
PubMed: 26798518
DOI: 10.1155/2015/648134 -
International Journal of Infectious... Jun 2015Breast implant infections are usually caused by Staphylococcus aureus and coagulase-negative staphylococci. Gram-negative bacilli are rarely reported to be involved in...
BACKGROUND
Breast implant infections are usually caused by Staphylococcus aureus and coagulase-negative staphylococci. Gram-negative bacilli are rarely reported to be involved in breast implant infections.
METHODS
Thirty-seven cases of microbiologically confirmed breast implant infection managed from January 2008 to June 2012 in the study centre were reviewed, including 10 cases from the study centre itself and 27 cases from private clinics in the region.
RESULTS
The prevalence of breast implant infection in the study centre was 0.74% of breast implantation, i.e., 3.23% in breast reconstruction for breast cancer and 0.27% in aesthetic breast augmentation (p=0.0002). Of the 37 cases, 30% had undergone radiotherapy and 11% had undergone a lymph node dissection. S. aureus was identified in 18 cases, Gram-negative bacilli in 10 cases, coagulase-negative staphylococci in eight cases, anaerobic bacteria in eight cases, and streptococci in three cases. Pseudomonas aeruginosa was the second most commonly identified pathogen. Staphylococcus epidermidis was the most frequent coagulase-negative Staphylococcus species. In addition to Propionibacterium acnes and Actinomyces neuii, other facultative and strict anaerobic bacteria have not been reported before, e.g., Bacteroides thetaiotaomicron, Corynebacterium simulans, Dermabacter hominis, Finegoldia magna, and Peptoniphilus harei. Seventy-percent of cases were treated by immediate implant removal. All cases treated only with antibiotics were treated with surgery at the second visit.
CONCLUSIONS
The microbiological epidemiology was noted by an increasing the proportion of Gram-negative bacteria and anaerobic bacteria detected with the advent of MALDI-TOF MS and molecular identification for diagnosis.
Topics: Adult; Aged; Bacteria; Bacterial Infections; Breast Diseases; Breast Implants; Female; France; Gram-Negative Bacteria; Humans; Mammaplasty; Middle Aged; Postoperative Complications; Pseudomonas aeruginosa; Referral and Consultation; Staphylococcus; Staphylococcus aureus; Young Adult
PubMed: 25910855
DOI: 10.1016/j.ijid.2015.04.010 -
PloS One 201316S rRNA-based genomic analyses have revolutionized our understanding of infectious diseases. Many cases which were recognized as "idiopathic" are now known to have an... (Comparative Study)
Comparative Study
BACKGROUND
16S rRNA-based genomic analyses have revolutionized our understanding of infectious diseases. Many cases which were recognized as "idiopathic" are now known to have an infectious etiology. Here, we present a proof-of-concept study to examine the microbial link between intra-amniotic infection (IAI) and early-onset neonatal sepsis (EONS).
RESULTS
Using culture independent methods, we analyzed paired amniotic fluid (AF) and cord blood (CB) samples from 36 singleton pregnancies complicated by preterm birth (PTB), IAI, and/or EONS. PTB cases were grouped as 1) Group 1- neonatal blood culture-positive EONS (n=6). 2) Group 2- neonatal blood culture-negative presumed EONS with positive IAI (n=16). 3) Group 3- neonatal blood culture-negative presumed EONS with no IAI (n=7); 4) Group 4- no EONS or IAI (n=7). In addition, samples from term healthy deliveries (n=8) served as technical controls. A total of 31 species (15 non-redundant) were identified in AF, of which only 1/3 were cultivated. Significantly fewer microorganisms were detected in CB, with a total of 18 species (7 non-redundant) identified, of which only 2 (Escherichia coli, Streptococcus agalactiae) were cultivated. Of those, Bergeyella, Fusobacterium nucleatum, and Sneathia sanguinegens had not been detected in EONS before. The novel species identified in AF by PCR include Peptoniphilus harei and Lachnospiraceae sp. The majority (72%) of CB species were also detected in the matching AF, with E. coli and F. nucleatum as the most prevalent. The 16S rRNA sequences of paired AF and CB were 99.9-100% identical, while no identical sequences were found between different pregnancies.
CONCLUSIONS
Previously unrecognized, uncultivated or difficult-to-cultivate species are implicated in EONS. Microbial species in paired AF and CB likely share the same infectious origin. Given its prevalence in EONS, F. nucleatum should be placed on the same importance scale as E. coli.
Topics: Adult; Amniotic Fluid; Bacteria; Base Sequence; Female; Fetal Blood; Humans; Infant, Newborn; Infant, Newborn, Diseases; Metagenome; Pregnancy; Premature Birth; RNA, Ribosomal, 16S; Sepsis; Species Specificity; Young Adult
PubMed: 23437088
DOI: 10.1371/journal.pone.0056131 -
APMIS : Acta Pathologica,... Apr 2008In the present study the lower genital tract microbiota in asymptomatic fertile women (n=34) was identified and quantified by culturing vaginal secretions. Also, vaginal... (Comparative Study)
Comparative Study
In the present study the lower genital tract microbiota in asymptomatic fertile women (n=34) was identified and quantified by culturing vaginal secretions. Also, vaginal and cervical samples were analyzed by a semiquantitative checkerboard DNA-DNA hybridization technique (CDH) based on genomic probes prepared from 13 bacterial species (Bacteroides ureolyticus, Escherichia coli, Fusobacterium nucleatum, Gardnerella vaginalis, Mobiluncus curtisii ss curtisii, Prevotella bivia, Prevotella disiens, Prevotella melaninogenica, Atopobium vaginae, Lactobacillus iners, Staphylococcus aureus ss aureus, Streptococcus anginosus, and Streptococcus agalactiae). The bacterial species found by either culture or CDH were correlated with proinflammatory cytokines (IL-1 alpha, IL-1 beta, IL-6, IL-8), secretory leukocyte protease inhibitor (SLPI), and endotoxin in the cervicovaginal samples. Grading the women into healthy, intermediate, or bacterial vaginosis (BV) as based on Gram staining of vaginal smears, the viable counts of lactobacilli (L. gasseri) and of streptococci-staphylococci combined were highest in the intermediate group. In BV, particularly the high concentrations of Actinomyces urogenitalis, Atopobium vaginae, and Peptoniphilus harei were noted (>or=10(11) per ml). The total viable counts correlated with both cervical IL-1 alpha and IL-1 beta. A strong negative correlation was observed between L. iners and total viable counts, G. vaginalis, or cervical IL-1 alpha, while it correlated positively with SLPI. Analysis of vaginal and cervical samples from 26 out of the 34 women by CDH showed that anaerobic bacteria were more frequently detected by CDH compared to culture. By this method, A. vaginae correlated with G. vaginalis, and L. iners with S. aureus. With regard to cytokines, B. ureolyticus correlated with both cervical and vaginal IL-1 alpha as well as with cervical IL-8, while F. nucleatum, S. agalactiae, S. anginosus, or S. aureus correlated with vaginal IL-1 alpha. Furthermore, all Gram-negative bacteria taken together, as measured by CDH, correlated with vaginal endotoxin and inversely with vaginal SLPI. The significance of the results is discussed. In summary, mapping of the identity and quantity of vaginal bacterial species and their association with locally produced host innate immune factors will help in defining various types of abnormal vaginal microbiota, developing new ways of assessing the risk of ascending subclinical infections, and in treating them. CDH appears to be a suitable tool for future analyses of large numbers of clinical samples with an extended number of bacterial probes.
Topics: Adult; Bacteria; Bacteriological Techniques; Carrier State; Cervix Mucus; Cervix Uteri; Colony Count, Microbial; Cytokines; DNA Probes; DNA, Bacterial; Endotoxins; Female; Humans; Middle Aged; Nucleic Acid Hybridization; Secretory Leukocyte Peptidase Inhibitor; Vagina; Vaginal Smears; Vaginosis, Bacterial
PubMed: 18397461
DOI: 10.1111/j.1600-0463.2008.00808.x -
Zhongguo Yi Xue Ke Xue Yuan Xue Bao.... Oct 2018Objective To investigate the associations of socioeconomic factors,nutrients intake,and gut microbiota of healthy pregnant women in the third trimester with gestational...
Objective To investigate the associations of socioeconomic factors,nutrients intake,and gut microbiota of healthy pregnant women in the third trimester with gestational weight gain (GWG).Methods We recruited 98 pregnant women in the third trimester who had received antenatal care in the Department of Obstetrics Gynecology,Peking Union Medical College Hospital from October,2015 to May,2016. We collected socioeconomic information through a structured questionnaire covering age,ethnicity,height,pre-pregnancy weight,and education. Nutritional status of these pregnant women was assessed by a 24-hour dietary intake recall. The participants were provided with collective tubes for faecal sample collection at home;their weight before the delivery was recorded. The pre-pregnancy weight and GWG were classified according to World Health Organization body mass index (BMI) standard for adults and the Institute of Medicine GWG guidelines (2009),respectively. The gut microbiota of the participants were analyzed using a whole-metagenome shotgun sequencing method.Results Insufficient and excessive GWG accounted for 15.3% and 50.0% of the cohort,respectively. Appropriate GWG level was associated with intakes of fat (F=3.113,P=0.049),carbohydrates (F=3.750,P=0.027),and dietary fiber (F=4.499,P=0.014) but not with age (F=2.495,P=0.088),ethnicity (Χ =0.065,P=0.968),education (Χ =0.827,P=0.661),or pre-pregnancy BMI (F=0.121,P=0.887). Compared with the participants with appropriate GWG,those with excessive GWG had significantly higher abundance of Akkermansia muciniphila,Atopobium parvulum,and Alistipes indistinctus as well as lower abundance of Lactobacillus rhamnosus,Weissella unclassified,Eubacterium ventriosum,Ruminococcus torques,and Bacteroides uniformis. Compared with the participants with appropriate GWG,those with insufficient GWG had significantly higher abundance of Dialister invisus,Alistipes unclassified,Peptoniphilus harei,Escherichia unclassified,Parvimonas unclassified,Campylobacter ureolyticus,Lactobacillus crispatus,and Fusobacterium nucleatum and lower abundance of Eubacterium ventriosum.Conclusions Abnormal GWG is common in pregnant women. GWG is significantly associated with gut microbiota as well as with nutritional factors including fat,carbohydrate,and dietary fiber intake.
Topics: Body Mass Index; Diet; Female; Gastrointestinal Microbiome; Gestational Weight Gain; Humans; Nutrients; Pregnancy; Pregnancy Trimester, Third; Socioeconomic Factors
PubMed: 30404694
DOI: 10.3881/j.issn.1000-503X.10505