Preliminary reading of antibiogram by microdilution for clinical isolates in urine culture.

PURPOSE: We evaluated a modification of automated antibiograms in urine cultures designed to facilitate the early interpretation of minimum inhibitory concentrations (MICs) and accelerate the targeted treatment of urinary tract infections (UTIs), METHODS: A prospective study was conducted of 309 isolates (219 Enterobacteriaceae, 75 Enterococcus spp., and 15 non-fermenting Gram-negative bacilli (NFGNB), and a retrospective study of 9 carbapenemase-producing clinical isolates from urine cultures. Colonies grown on conventional isolation plates were inoculated in MicroScan Walkaway system panels and incubated for 7 h, using a MicroScan AutoScan-4 plate reader for preliminary MIC determination by turbidimetry. Resulting antibiograms were compared with definitive antibiograms obtained after incubation for 17 h. RESULTS: Preliminary and definitive readings were concordant for 86.7% of Gram-positive cocci isolates (65/75), 61.6% of Enterobacteriaceae (135/219), and 53.3% of NFGNB. The agreement rate was greater than 90% for most antimicrobials against Gram-positive cocci (94.7% or more) and Enterobacteriaceae, (97.2% or more for 10 of 17 antibiotics) except with nitrofurantoin (89%). The agreement rate was 86.7% or more for most antibiotics against NFGNB apart from piperacillin/tazobactam, aztreonam, amikacin, and ciprofloxacin. Gram-negative bacilli showed the highest differences in MIC values between preliminary and definitive readings. CONCLUSIONS: A preliminary antibiogram reading may be useful in urine cultures to reduce the delay before targeted antibiotherapy, especially against Enterobacteriaceae and Gram-positive cocci, but not in cases of carbapenemase-producing NFGNB. Further local studies are warranted to evaluate the usefulness of this approach in relation to resistance rates.

Reliability of a Screening Method Using Antibiotic Disks to Detect Carbapenemases in Glucose-Nonfermenting Gram-Negative Microorganisms From Clinical Samples of a Regional Hospital in Southeastern Spain.

BACKGROUND: Infections by glucose-nonfermenting gram-negative bacilli (NFGNB) pose a major public health problem due to multiresistance to beta-lactam antibiotics, especially plasmid-borne carbapenemases. Their detection by microbiology laboratories is challenging, and there is a need for easy-to-use and reliable diagnostic techniques. Our objective was to evaluate an in-house screening method to presumptively detect carbapenemases in NFGNB in a simple and clinically useful manner. METHODS: The study included 175 NFGNB isolates from urinary, respiratory, and rectal samples. In a triple assay, isolates were incubated at 37°C for 24 h on three solid-culture media: MacConkey II Agar, 5% Sheep Blood Columbia Agar and Mueller Hinton II Agar; meropenem (MEM) and cefepime (FEP) disks were employed for screening. Studies were then performed on the inhibition halo diameter, scanning effects, and the appearance of mutant colonies, which were compared with those observed using the colorimetric Neo-Rapid CARB Kit and immunochromatography (NG5-Test Carba and K-Set for OXA-23). Receiver operating characteristic curves were constructed for these data. RESULTS: Carbapenemases were expressed by 79/175 (45.1%): 19 Pseudomonas aeruginosa and 60 Acinetobacter baumannii. Optimal inhibition halo diameter cutoffs to detect this resistance on 5% sheep blood agar were as follows: 6 mm (MEM) and 6.5 mm (FEP) for P. aeruginosa (in the absence of scanning effects and mutations) and 10.5 mm (MEM) and 16 mm (FEP) for A. baumannii (even in the presence of scanning effects). CONCLUSION: The combined utilization of MEM and FEP antibiotic disks in 5% sheep blood agar, measuring their inhibition haloes, offers an effective method to predict the presence of carbapenemases as resistance mechanism in P. aeruginosa and A. baumannii.

Bacteremia caused by Veillonella parvula: Two case reports and a review of the literature.

Veillonella parvula is a non-motile Gram-negative coccus that forms part of the normal microbiota in several body sites and which has been rarely isolated as cause of infections in human population, particularly in bacteremias. Here we give the overview of characteristics of genus Veillonella and the summary of its role in infections, particularly in bacteremia. We additionally report two patients with bacteremia due to V. parvula. Two sets of blood cultures of each patient yielded a pure culture of an anaerobic microorganism identified as V. parvula by MALDI-TOF MS, and confirmed by 16S rRNA gene sequencing. The two patients were male and one of them had risk factors for anaerobic bacteremia. The isolates were susceptible to most antibiotics and the outcome was successful in both patients. Bacteremia due to V. parvula is still rare. MALDI-TOF MS appear to be an excellent tool for the correct identification of these species.

In vitro activity of delafloxacin against anaerobic bacteria compared with other antimicrobials.

The aim of this study was to describe the in vitro activity of delafloxacin against 230 anaerobic isolates and compare it with the activity of other antimicrobials used against infections caused by anaerobic microorganisms. Minimal inhibitory concentrations (MICs) were lower for delafloxacin than for all other antibiotics tested with the exception of piperacillin-tazobactam and meropenem against Gram-positive anaerobic cocci. Only two (0.8 %) isolates of Bacteroides spp. showed a MIC ≥4 µg/mL. With some exceptions, the present results show lower MICs for delafloxacin in comparison to the other antibiotics used against anaerobes.

New anaerobic bacteria causing infection: Bacteremia due to Murdochiellaasaccharolytica.

Murdochiella asaccharolytica is a Gram-positive anaerobic coccus firstly isolated from wound specimens, since then, it was very rarely isolated from human infections. We report the first case of bloodstream infection due to this bacterium in a 90 years-old-man. The patient had a history of B-cell lymphoma and presented with pain and induration of the base of penis along with fever. A blood culture was obtained yielding M. asaccharolytica in pure culture. Only resistance to clindamycin was demonstrated. Treatment with metronidazole was started and the clinical evolution improved with antimicrobial treatment, being discharged after 6 days.

Comparative evaluation of MALDI-TOF MS and 16S rRNA gene sequencing for the identification of clinically relevant anaerobic bacteria: critical evaluation of discrepant results.

OBJECTIVES: The main study objective was to evaluate the correlation between matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequencing results for the identification of anaerobes. METHODS: A retrospective study was conducted of all anaerobic bacteria isolated from clinically significant specimens. MALDI-TOF (Bruker Byotyper) and 16S rRNA gene sequencing were performed in all strains. Identifications were considered correct when the concordance with gene sequencing was ≥99%. RESULTS: The study included 364 isolates of anaerobic bacteria: 201 (55.2%) Gram-negative and 163 (44.8%) Gram-positive, mostly belonging to the genus Bacteroides. Isolates were largely obtained from blood cultures (128/35.4%) and intra-abdominal samples (116/32.1%). Overall, 87.3% of isolates were identified at species level using the version 9 database (89.5% of Gram-negative and 84.6% of Gram-positive anaerobic bacteria). All isolates belonging to the species B. fragilis sensu stricto were correctly identified by MALDI-TOF MS, but five cases of Phocaeicola (Bacteroides) dorei were misidentified as Phocaeicola (Bacteroides) vulgatus; all Prevotella isolates were correctly identified at the genus level, and most were correctly identified at the species level. Among Gram-positive anaerobes, 12 Anaerococcus species were not identified by MALDI-TOF MS, while six cases identified as Peptoniphilus indolicus were found to belong to other genera/species. CONCLUSIONS: MALDI-TOF is a reliable technique for identifying most anaerobic bacteria, although the database needs frequent updating to identify rare, infrequent, and newly discovered species.

Bacteremia due to Clostridium innocuum: Analysis of four cases and literature review.

Clostridium innocuum is a Gram-positive anaerobic spore-forming bacillus that has been identified as part of the normal intestinal microbiota. This bacterium has been rarely associated with human infections, and only few severe infections have been reported until now. In this work, we report on four patients with bacteremia due to C. innocuum, which were well identified by MALDI-TOF MS. Moreover, a review of the previous published cases of bacteremia due to this anaerobic bacterium has been performed.

Differences between bloodstream infections involving Gram-positive and Gram-negative anaerobes.

BACKGROUND: The objectives of this study were to describe differences between bloodstream infections involving Gram-positive (GP) and Gram-negative (GN) anaerobic bacteria. METHODS: Patients with clinically significant anaerobic bacteremia detected between October 2016 and July 2022 in a tertiary hospital in Granada (Spain) were retrospectively included. Species identification was performed by MALDI-TOF MS and/or molecular methods. The association between variables was analyzed using contingency tables, applying the chi-square test when expected frequencies were adequate and the Fisher exact test when not. Variables were gathered at the time of the first positive blood culture. RESULTS: Out of 237 cases of anaerobic bloodstream infections detected, 127 (53.6%) were GN. Crude mortality was 20.3%, corresponding to 48 patients who died of causes directly attributable to bacteremia. The presence of malignant disease (p = 0.011), abdominal and/or pelvic surgery (p = 0.001), and transplantation (p = 0.008) were significantly associated with bacteremia due to GN bacteria, while the presence of diabetes mellitus was significantly associated with bacteremia due to GP bacteria (p = 0.022). The presence of both septic shock and mortality was more frequently associated with bacteremia due to GN versus GP bacteria. CONCLUSIONS: The association of certain variables with the presence of bloodstream infections due to GP or GN anaerobic bacteria may assist in selecting the optimal empirical therapeutic approach and improving the outcome of patients with these types of infection.

An uncommon case of bacteremia caused by Lancefieldella parvula in an oncological patient.

We report an uncommon case of bacteremia due to Lancefieldella parvula isolated in pure culture. We present a 72 year-old-man affected with cancer admitted with abdominal pain due to a parietal rupture of the urinary bladder. After surgical treatment, the patient developed fever and two sets of blood cultures yielded a pure culture of an anaerobic microorganism identified as L. parvula (formerly Atopobium parvulum) by MALDI-TOF MS, and confirmed by 16S rRNA gene sequencing. The isolate was susceptible to all antibiotics and the outcome was finally successful. Bacteremia due to L. parvula is an uncommon disease and, in that case, MALDI-TOF MS was an useful tool for the initial identification.

Brain abscesses caused by anaerobic bacteria.

OBJECTIVES: The objective of this study was to review the characteristics, causative bacteria, treatment and outcomes of brain abscesses due to anaerobes in our health setting. METHODS: A retrospective analysis of all brain abscesses caused by anaerobic bacteria over the period 2005-2021 was performed. RESULTS: Out of 300 brain abscesses identified during the study period, 31 were produced by anaerobic pathogens, either alone (monomicrobial infection) or together with aerobic and/or anaerobic bacteria (polymicrobial infection). The mean age of the 31 patients was 53 years, and 61.2% were male; 51.6% of infections were polymicrobial, with only four (12.9%) caused by anaerobic bacteria alone. Forty-three anaerobic bacteria were isolated: Cutibacterium acnes in thirteen (41.9%), Parvimonas micra in eight (25.8%), and Prevotella spp. in seven (22.5%). The most frequent etiologies were local neurosurgery (13/41.9%) and contiguous otogenic, oral, or sinus foci of infection (8/28.8%). Cancer was present in eight patients (28.8%), headaches in seventeen (54.8%), and fever in nine (28.8%). All patients received both surgery and antimicrobial therapy. The abscess was in the frontal region in 12 patients (38.7%) and in the parietal region in 11 (35.4%). A good outcome was obtained in all patients. CONCLUSIONS: Anaerobes were isolated in 10.3% of patients with brain abscesses in our health setting, similar to other reports. C. acnes was the most frequently detected anaerobe, especially in neurosurgical patients.

Bacteremia due to Prevotella oris of probable hepatic origin.

Prevotella species are Gram-negative anaerobic bacilli mainly involved in oral cavity infections. We report a rare case of bacteremia due to P. oris of probable hepatic origin. A 70-year-old man with a history of diabetes mellitus was admitted for general malaise, fever and dyspnea. The patient was admitted to the ICU due to septic shock. Two sets of blood cultures yielded a pure culture of an anaerobic microorganism identified as P. oris by MALDI-TOF MS. The same microorganism was isolated from a hepatic abscess drainage. Only resistance to penicillin was documented. Treatment with piperacillin-tazobactam and levofloxacin was administered, and the patient was discharged following his recovery.

Bacteremia due to Parabacteroides distasonis: Experience with 4 cases.

OBJECTIVE: To report a series of four uncommon cases of bacteremia due to Parabacteroides distasonis. METHODS: We have analyzed the epidemiological characteristics, clinical features, diagnostic methods, treatment and outcome of these patients. RESULTS: Two sets of blood cultures of each patient yielded a pure culture of an anaerobic microorganism identified as P. distasonis by MALDI-TOF MS, and confirmed by 16S rRNA gene sequencing. All patients were male and they had risk factors for anaerobic bacteremia. All isolates were susceptible to metronidazole and the outcome was successful in three patients. CONCLUSIONS: Bloodstream infections due to P. distasonis are still rare. MALDI-TOF MS appear to be an excellent tool for the correct identification.

Misidentification of Phocaeicola (Bacteroides) dorei in two patients with bacteremia.

Phocaeicola (Bacteroides) dorei is a Gram-negative anaerobic bacillus that is rarely isolated from human specimens. Its accurate identification can be hampered by its close taxonomic relationship with Bacteroides vulgatus. We report on two patients with bacteremia due to P. (B.) dorei, which was initially identified as B. vulgatus by MALDI-TOF MS.

Curvicollide D Isolated from the Fungus Amesia sp. Kills African Trypanosomes by Inhibiting Transcription.

Sleeping sickness or African trypanosomiasis is a serious health concern with an added socio-economic impact in sub-Saharan Africa due to direct infection in both humans and their domestic livestock. There is no vaccine available against African trypanosomes and its treatment relies only on chemotherapy. Although the current drugs are effective, most of them are far from the modern concept of a drug in terms of toxicity, specificity and therapeutic regime. In a search for new molecules with trypanocidal activity, a high throughput screening of 2000 microbial extracts was performed. Fractionation of one of these extracts, belonging to a culture of the fungus Amesia sp., yielded a new member of the curvicollide family that has been designated as curvicollide D. The new compound showed an inhibitory concentration 50 (IC50) 16-fold lower in Trypanosoma brucei than in human cells. Moreover, it induced cell cycle arrest and disruption of the nucleolar structure. Finally, we showed that curvicollide D binds to DNA and inhibits transcription in African trypanosomes, resulting in cell death. These results constitute the first report on the activity and mode of action of a member of the curvicollide family in T. brucei.

Different breakpoints interpretation yielded distinct resistance rates to moxifloxacin of clinically significant anaerobic bacteria.

The aim of this study was to describe the differences in antimicrobial susceptibility to moxifloxacin between European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Clinical and Laboratory Standards Institute (CLSI) in anaerobic microorganisms. Overall, resistance to moxifloxacin appears to be high in almost all groups of anaerobes, but enormous differences in susceptibility rates between these two committees could be observed.

Bacteremia caused by Anaerococcus SPP: Is this an underdiagnosed infection?

The objectives of this study were to report 10 episodes of clinically significant bacteremia caused by species of the genus Anaerococcus isolated between July 2018 and February 2021 from the microbiology laboratory of a tertiary hospital in Granada (Spain). None of the isolates were identified by MALDI-TOF MS, and the definitive species identification was performed by 16 S rRNA gene sequencing. No reference spectra of the Anaerococcus species were present in the MALDI-TOF MS database. Eight isolates were finally identified as A. octavius, one isolate as A. tetradius and the other as A. urinomassiliensis. The majority of these infections were seen in patients aged >70 years. Risk factors for anaerobic infection were observed in eight patients, especially diabetes mellitus, surgery, and the presence of cancer. Fever was present in all patients. Three patients died, but only one death was attributed to the infection. Mean detection time of positive blood cultures was 47.5 h (range 24-92 h). Antimicrobial susceptibility to penicillin, amoxicillin-clavulanate, imipenem, moxifloxacin, clindamycin, metronidazole, and piperacillin-tazobactam was tested using the gradient diffusion technique and EUCAST breakpoints (except for moxifloxacin). No resistance to amoxicillin-clavulanate, metronidazole, imipenem, or piperacillin-tazobactam was detected; however, the majority of isolates were resistant to clindamycin. When MALDI-TOF MS does not provide a correct identification at genus or species level, as in some isolates of Gram-positive anaerobic cocci, microbiologists should perform an additional confirmatory technique, such as gene sequencing analysis, to obtain a definitive diagnosis.

A rare cause of bacteremia due to Porphyromonas asaccharolytica in a patient with necrotizing fasciitis.

Porphyromonas species are Gram-negative anaerobic bacilli mainly involved in human periodontal diseases. We report an uncommon case of bacteremia due to P. asaccharolytica in a patient with necrotizing fasciitis. A 52-year-old woman with a history of diabetes mellitus was admitted for an extensive necrotizing lesion on the left lower limb. After she developed septic shock, two sets of blood cultures were taken. Anaerobic bottles yielded a pure culture of a microorganism initially identified as P. uenonis by MALDI-TOF MS but with a low log score, and a gene sequencing technique was therefore applied, identifying the isolate as P. asaccharolytica. Only resistance to penicillin and clindamycin was documented. Treatment with meropenem was administered, and the patient was discharged following her recovery.

First case of abdominal infection caused by bacteroides fluxus.

Bacteroides fluxus is a Gram-negative anaerobic bacillus isolated from human faeces in healthy individuals. Until now, this bacterium had not been involved in human diseases. We report the first case of abdominal infection due to this microorganism in an elderly patient. A 76-year-old man with a history of chronic pulmonary obstructive disease presented with dyspnea, orthopnea and cough. The clinical evolution worsened with both a colonic ischemia and further diffuse peritonitis of pancreatic origin. Peritoneal fluid was obtained and the culture yielded B. fluxus in pure culture. Resistance to penicillin, amoxicillin-clavulanate, clindamycin and moxifloxacin was documented. Treatment with meropenem + linezolid was started, but the patient finally died due to a multiorganic failure.

Recent clinical relevance of mono-genital colonization/infection by Ureaplasma parvum.

Ureaplasma parvum is the most prevalent genital mycoplasma in women of childbearing age. There is debate around the relevance of its presence in male or female genitals for disease development and as a cofactor. The objective of this study was to determine the prevalence of colonization/infection by U. parvum and its possible relationship with reproductive tract infections. We retrospectively analyzed the presence of U. parvum in patients referred by specialist clinicians for suspicion of genitourinary tract infection. U. parvum was detected in 23.8% of samples, significantly more frequently in females (39.9%) than in males (6%). Among the males, U. parvum was found alone in 68.4% of episodes, with Ct < 30. Among the females, U. parvum was detected in 88.6% of cases, with Ct < 30, including 22 cases with premature rupture of membranes and 6 cases with threat of preterm labor. Co-infection was significantly more frequent in females (62.6%) than in males (31.6%). Given the high prevalence of U. parvum as sole isolate in males and females with genitourinary symptoms, it should be considered in the diagnosis and treatment of genital infections, although its pathogenic role in some diseases has not been fully elucidated.

Potential clinical use of azithromycin against gastroenteritis-causing pathogens other than Campylobacter.

The activity of azithromycin against enteritis-producing agents other than Campylobacter spp. was studied. The susceptibility to azithromycin, through gradient test, of 88 clinical isolates (51 Salmonella spp., 23 Aeromonas spp., 10 Shigella sonnei and 4 Yersinia enterocolitica) for one year was studied prospectively. The results were compared with the activity of ampicillin, trimethoprim-sulfamethoxazole and ciprofloxacin by microdilution. For azithromycin, the minimum inhibitory concentration (MIC) 50 and MIC90 were 4 and 12 mg/l, respectively. Six (6.8%) isolates were simultaneously resistant to ampicillin, trimethoprim- sulfamethoxazole and ciprofloxacin, and 3 (50%) of them presented a MIC >256 mg/l. Azithromycin may be a good empirical therapeutic option for the treatment of bacterial enteritis.