Microbiological Samples Research Articles

Devices for the transport and microbiological detection of Neisseria gonorrhoeae in sexual health clinic samples: a prospective comparative study.

Propagation and microbiological detection of N. gonorrhoeae can be challenging due to the fastidious nature of the bacterium outside the human host environment. Different sample transport options were evaluated in order to address this issue. Symptomatic and high-risk patients consented to take part. Standard practice in clinics involve using a guanine and cytosine selective (GC) plate for transport and incubation. Other devices were assessed in two different studies. MWE's Sigma VCM™-a transport device for use between clinic and laboratory-was used in one study (n = 166). In the other, (n = 102), Biomed's Intray™ plates were used, which can be used for both initial transport and incubation at the destination laboratory. In the Sigma VCM™ study, positive results were obtained for 14% (Sigma VCM™), 21% (GC plates); the distribution of outcomes did not significantly differ between the two microbiological sampling methods (p-value 0.09, Chi-squared test). Concerning the InTray™ sub-study, N. gonorrhoeae was detected in 9% (InTray™), 13% (GC plating) of cases respectively, with again no significant difference between the microbiological sampling methods (p-value 0.25). Regression analysis identified a significant association between N. gonorrhoeae detection and male patients, absence of dysuria and previous gonorrhoea infection. No significant difference in rate of microbiological detection of N. gonorrhoeae could be detected between different transport devices in a sexual health clinic setting. Performance appraisal of transport devices for bacterial sexually transmitted infection can inform practitioners' options regarding said devices.

Invasive Aggregatibacter infection: shedding light on a rare pathogen in a retrospective cohort analysis.

Introduction. Aggregatibacter are Gram-negative, facultatively anaerobic rods or coccobacilli that are infrequently encountered as pathogens causing infection.Hypothesis/Gap Statement. The range of invasive infection that Aggregatibacter cause is poorly described. The pathogenicity of species such as Aggregatibacter segnis is debated.Aim. To identify invasive infection due to Aggregatibacter species in a large healthcare organization and to characterize clinical syndromes, co-morbidities and risk factors.Methodology. All microbiological samples positive for Aggregatibacter species were identified by conventional culture or 16S rRNA PCR between October 2017 and March 2021. Electronic records for all patients with positive samples were reviewed and the infection syndrome classified for patients with invasive disease.Results. Twenty-seven patients with invasive infection were identified, with a statistically significant difference in species-specific patterns of invasive infection (P=0.02) and a statistically significant association with residence in the 30 % most deprived households in the UK by postcode (P<0.01). The three most common co-morbidities were periodontitis or recent dental work (29.6%), cardiovascular disease (25.9%) and diabetes (18.5 %).Conclusion. We describe a novel association of Aggregatibacter segnis with skin and soft tissue infection. The propensity of the Aggregatibacter species to cause invasive infection at different body sites and be associated with deprivation is reported. Aggregatibacter actinomycetemcomitans bacteraemia was associated with infective endocarditis, and Aggregatibacter aphrophilus was implicated in severe appendicitis and noted to cause brain abscess. Areas warranting future research include exploring the risk-factors required for invasive infection and those that may determine the species-specific differences in patterns of invasive disease.

Antimicrobial Resistance Profile of Bacteria Causing Pediatric Infections at the University Teaching Hospital in Rwanda.

Bacterial infections pose a global threat, especially in the pediatric population. Antimicrobials that are used to treat such infections continuously show reduced efficacy, and empirical therapy is a major treatment option in Rwanda. This study aimed to determine the resistance rate of commonly used antibiotics in pediatric patients. The study was conducted from June 1, 2018 to May 30, 2019, and microbiological samples were collected from 712 children with suspected bacterial infections. Antimicrobial sensitivity testing was performed on 177 positive cultures (24%) that were considered for data analysis. The findings show that the major bacterial isolates were Klebsiella pneumoniae (n = 50, 28.2%), Escherichia coli (n = 47, 26.5%), and Staphylococcus aureus (n = 38, 21.4%). In general, the greatest antibiotic resistance rate was observed in ampicillin (n = 125, 86.2%), amoxicillin-clavulanic acid (n = 84, 82.4%), amoxicillin (n = 64, 79%), cefadroxil (n = 83, 69.2%), tetracycline (n = 72, 59.7%), ceftazidime (n = 42, 55.3%), and cefuroxime (n = 14, 53.8%). More specifically, Klebsiella pneumoniae was 100% resistant to amoxicillin-clavulanic acid, cefuroxime, trimethoprim-sulfamethoxazole, ceftazidime, erythromycin, and clindamycin. Staphylococcus aureus was 86.7% resistant to ampicillin, and Escherichia coli was 91.7% resistant to tetracycline, 90.6% resistant to ampicillin, 83.3% resistant to amoxicillin-clavulanic acid, 79.3% resistant to cefadroxil, and 78.6% resistant to ceftazidime. Moreover, Klebsiella pneumoniae from blood and urine was 96.8% and 100% sensitive, respectively, to meropenem. Staphylococcus aureus from blood was 100% sensitive to vancomycin, whereas Escherichia coli from urine was sensitive to clindamycin (100%), nitrofurantoin (80.6%), and ciprofloxacin (72.7%). In conclusion, our findings show a high resistance rate to commonly used antibiotics, which suggests precaution in empirical therapy and continued surveillance of antimicrobial resistance.

Case-control investigation of invasive Salmonella disease in Malawi reveals no evidence of environmental or animal transmission of invasive strains, and supports human to human transmission.

Invasive Salmonella infections cause significant morbidity and mortality in Sub-Saharan Africa. However, the routes of transmission are uncertain. We conducted a case-control study of index-case and geographically-matched control households in Blantyre, Malawi, sampling Salmonella isolates from index cases, healthy people, animals, and the household environment. Sixty index cases of human invasive Salmonella infection were recruited (March 2015-Oct 2016). Twenty-eight invasive Non-Typhoidal Salmonella (iNTS) disease and 32 typhoid patients consented to household sampling. Each index-case household was geographically matched to a control household. Extensive microbiological sampling included stool sampling from healthy household members, stool or rectal swabs from household-associated animals and boot-sock sampling of the household environment. 1203 samples from 120 households, yielded 43 non-Typhoidal Salmonella (NTS) isolates from 25 households (overall sample positivity 3.6%). In the 28 iNTS patients, disease was caused by 3 STs of Salmonella Typhimurium, mainly ST313. In contrast, the isolates from households spanned 15 sequence types (STs). Two S. Typhimurium isolates from index cases closely matched isolates from their respective asymptomatic household members (2 and 3 SNP differences respectively). Despite the recovery of a diverse range of NTS, there was no overlap between the STs causing iNTS disease with any environmental or animal isolates. The finding of NTS strains from index cases that matched household members, coupled with lack of related animal or environmental isolates, supports a hypothesis of human to human transmission of iNTS infections in the household. The breadth of NTS strains found in animals and the household environment demonstrated the robustness of NTS sampling and culture methodology, and suggests a diverse ecology of Salmonella in this setting. Healthy typhoid (S. Typhi) carrier state was not detected. The lack of S. Typhi isolates from the household environment suggests that further methodological development is needed to culture S. Typhi from the environment.

Bacterial keratitis – Challenges with changing microbial resistance

AbstractAbout 90% of corneal infections are bacterial in origin. The most common pathogens are Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae and Pseudomonas aeruginosa. The two main causes of bacterial keratitis are contact lens usage and corneal injury.Patients typically complain of pain, sensitivity to light, blurred vision, tearing and yellowish purulent discharge. Clinical signs of bacterial keratitis include epithelial defect, ulceration in addition, stromal oedema and infiltrates with indistinct margins are present. Purulent material may appear in the anterior chamber.Most cases can be successfully treated with empiric therapy. If the pathogen is not known, moxifloxacin or cefazolin with tobramycin or gentamycin recommended as primary treatment. For severe keratitis eye drops should be applied both day and night for the first 48 hours. If the causative agent and its susceptibility are available from the microbiological sample and/or from smear, the topical treatment should be adjusted accordingly. If no improvement in the clinical picture seen after 48 hours with topical treatment, repeat sampling for microbiological culture recommended. In case of further progression, keratectomy with histological evaluation of the specimen is necessary. Corticosteroids may be considered after 48 hours when the infective organism is identified, and the keratitis is responding to therapy.In addition to antimicrobial treatment pupillary dilation advised to immobilize the iris and ciliary body. Cycloplegia may also decrease pain as well as synechia formation. In therapy resistant cases crosslinking may be an additional option if herpetic origin can be excluded.If the clinical picture does not improve, it is important to reassess the differential diagnosis of the keratitis caused by herpetic, fungal and Acanthamoeba infection.In cases of non‐healing and rapidly progressive corneal inflammation or ulceration, therapeutic corneal transplantation is recommended. However, surgery has a better prognosis in the non‐inflammatory period when the eye is quiet.

A comparison of methods for microbiologic environmental sampling.

We compared the effectiveness of 4 sampling methods to recover Staphylococcus aureus, Klebsiella pneumoniae and Clostridioides difficile from contaminated environmental surfaces: cotton swabs, RODAC culture plates, sponge sticks with manual agitation, and sponge sticks with a stomacher. Organism type was the most important factor in bacterial recovery.

New Antibiotic Prescription Pattern in Critically Ill Patients ("Ant-critic"): Prospective Observational Study from an Indian Intensive Care Unit.

This study aimed to address the issue of antibiotic prescription processes in an Indian Intensive care unit (ICUs). In a prospective longitudinal study, all adult patients admitted in the ICU for 24 hours or above between 01 June 2020 and 31 July 2021 were screened for any new antibiotic prescription throughout their ICU stay. All new antibiotic prescriptions were assessed for baseline variables at prescription, any modifications during the course, and the outcome of antibiotic prescription. A total of 1014 patients fulfilled entry criteria; 59.2 and 7.2% of days they were on a therapeutic and prophylactic antibiotic(s). Patients, who were prescribed therapeutic antibiotic(s), had worse ICU outcomes. A total of 49.5% of patients (502 of 1,014) received a total of 552 new antibiotic prescriptions during their ICU stay. About 92.13% of these prescriptions were empirical and blood or other specimens were sent for culture in 78.81 and 60.04% of instances. A total of 31.7% of episodes were microbiologically proven and were more likely to be prescribed by an ICU consultant. A total of 169 modifications were done in 142 prescription episodes; 73 of them after sensitivity results. Thus, the overall rate of de-escalation was 13.95%. Apart from the negative culture result (36.05%), an important reason for a relatively low rate of de-escalation was the absence of sampling (12.32%). Longer ICU stay before antibiotic prescription, underlying chronic liver disease (CLD), worse organ dysfunction, and septic shock were independently associated with unfavorable treatment outcomes. No such independent association was observed between antibiotic appropriateness and patient outcome. Future antibiotic stewardship strategies should address issues of high empirical prescription and poor microbiological sampling hindering the de-escalation process. Ghosh S, Salhotra R, Singh A, Lyall A, Arora G, Kumar N, et al. New Antibiotic Prescription Pattern in Critically Ill Patients ("Ant-critic"): Prospective Observational Study from an Indian Intensive Care Unit. Indian J Crit Care Med 2022;26(12):1275-1284.

Clinical Microbial Identification of Severe Oral Infections by MALDI-TOF Mass Spectrometry in Stockholm County: an 11-Year (2010 to 2020) Epidemiological Investigation.

Growing evidence suggests that oral infections can modify the course of systemic diseases. To date, epidemiological data on microbial oral infections are scarce. Here, we performed a comprehensive analysis of the trend and microbial diversity in oral infection specimens referred for clinical microbiology analysis from 2010 to 2020. The microbes were isolated by culture and were identified via matrix-assisted laser desorption ionization-time of flight mass spectrometry technology (MALDI-TOF MS) throughout the study period. A total of 1,014 referred samples from dental clinics in Stockholm County with dentoalveolar abscesses and jaw osteomyelitis being the main reason were identified. Overall, the microbial composition was dominated by Firmicutes (51%), followed by Bacteroidetes (19%), Proteobacteria (12%), and Actinobacteria (5%). At the genus level, Streptococcus spp. (36%), Prevotella spp. (18%), and Staphylococcus spp. (11%) were among the most frequently reported. Interestingly, a strong increase in trend was noted for Streptococcus anginosus, Streptococcus mitis, Streptococcus sanguinis, Eikenella corrodens, Actinomyces spp., Aggregatibacter aphrophilus, Staphylococcus epidermidis, and Granulicatella adiacens during the study time (R = 0.66 to 0.89, P < 0.05), and a minor increase was noted for Enterococcus faecalis and Klebsiella spp., whereas steady levels were noted for most of the others. The present study shows the diversity of bacteria that have been involved in dental infections during the last decade in the capital of Sweden, as well as the emerging oral microbiota trend, with clear clinical implications on the oral-systemic link. IMPORTANCE Oral diseases and associated microbes are a risk factor for systemic diseases and can change the courses of these diseases. To date, epidemiological data on microbial oral infections are scarce, and longitudinal reports are lacking. We present for the first time the microbial composition of severe oral bacterial infections determined via the MALDI-TOF mass spectrometry technique in a comprehensive study between 2010 and 2020 (11 years) in Stockholm County. The trend and microbial diversity of oral infections were analyzed on referred clinical microbiological samples and were processed by standardized protocols. Trend increase was noted for Streptococcus anginosus, Streptococcus mitis, Streptococcus sanguinis, Eikenella corrodens, Actinomyces spp., Aggregatibacter aphrophilus, Staphylococcus epidermidis, Granulicatella adiacens, Enterococcus faecalis, and Klebsiella spp. Our results provide new insights into the diversity and trend of oral microbiota that were involved in serious oral infections over the past decade in the capital of Sweden and may influence the oral-systemic link.

Hygiene of Medical Devices and Minimum Inhibitory Concentrations for Alcohol-Based and QAC Disinfectants among Isolates from Physical Therapy Departments.

Disinfectants are used intensively to control and prevent healthcare-associated infections. With continuous use and exposure to disinfectants, bacteria may develop reduced susceptibility. The study aimed to check the hygiene of devices in the physiotherapy department. For isolated bacterial strains, we aimed to determine the minimum inhibitory concentration of five different disinfectant wipe products currently in use. Microbiological environmental sampling in four various institutions in four different cities from two counties was performed, followed by CFU calculation and identification using matrix-assisted laser desorption and ionization with time-of-flight analyzer mass spectrometry (MALDI-TOF). The sampling was performed on three different occasions: before patient use, after patient use, and after disinfection. The susceptibility of isolates to three different alcohol-based and three different quaternary ammonium compounds (QAC) disinfectant wipes was examined by determining the minimal inhibitory concentrations (MIC). We identified 27 different bacterial species from 11 different genera. Gram-positive bacteria predominated. The most abundant genera were Staphylococcus, Micrococcus, and Bacillus. The average MIC values of alcohol-based disinfectants range between 66.61 and 148.82 g/L, and those of QAC-based disinfectants range between 2.4 and 3.5 mg/L. Distinctive strains with four-fold increases in MIC values, compared to average values, were identified. The widespread use of disinfectants can induce a reduction in the susceptibility of bacteria against disinfectants and affect the increase in the proportion of antibiotic-resistant bacteria. Therefore, it is urgent to define clear criteria for defining a microorganism as resistant to disinfectants by setting epidemiological cut-off (ECOFF) values and standardizing protocols for testing the resistance of microorganisms against disinfectants.

Acquisition of extended-spectrum cephalosporin-resistant Gram-negative bacteria: epidemiology and risk factors in a 6-year cohort of 507 severe trauma patients

Severe trauma patients are at higher risk of infection and often exposed to antibiotics, which could favor acquisition of antimicrobial resistance. In this study, we aimed to assess prevalence, acquisition, and factors associated with acquisition of extended-spectrum cephalosporin-resistant Gram-negative bacteria (ESCR-GNB) in severe trauma patients. We conducted a retrospective monocentric cohort study in a French level one Regional Trauma Centre between 01 January 2010and 31 December 2015. Patients admitted for ≥ 7 days, with an Injury Severity Score ≥ 15, and ≥ 1 microbiological sample were included in the analysis. Prevalence and acquisition rate of ESCR-GNB were determined then, factors associated with ESCR-GNB acquisition were assessed using a Cox model. Of 1873 patients admitted during the study period, 507 were included (median Injury Severity Score=29 [22-34] and median intensive care unit length of stay=16 days [10-28]). Most of them (450; 89%) had an antimicrobial therapy. Prevalence of ESCR-GNB increased from 13% to 33% during intensive care unit stay, bringing the ESCR-GNB acquisition rate to 29%. Acquisition of ESCR-GNB was mainly related to AmpC beta-lactamase Enterobacterales and was independently associated with mechanical ventilation needs (hazard ratio [HR]=6.39; 95% confidence interval [CI] [1.51-27.17]; P=0.01), renal replacement therapy needs (HR=2.44; 95% CI [1.24-4.79]; P=0.01), exposure to cephalosporins (HR=1.06; 95% CI [1.01-1.12]; P=0.02), and/or combination therapy with non-beta-lactam antibiotics such as vancomycin, linezolid, clindamycin, or metronidazole (HR=1.03; 95% CI [1.01-1.06]; P=0.02). Acquisition of ESCR-GNB was prevalent in severe trauma patients. Our results suggest selecting antibiotics with caution, particularly in the most severely ill.

Prise en charge médico-chirurgicale des ostéites, point de vue de l’infectiologue et du chirurgien

AbstractOsteitis is the result of contamination of the bone by one or more microorganisms that cause bone destruction. The diagnostic approach includes two steps: positive diagnosis (based on clinical, biological and imaging findings) and microbiological diagnosis. The management will be medical-surgical. it is recommended to discuss the management in a multidisciplinary consultation meeting, or even in complex cases to present the case to a reference center for complex osteoarticular infections (Crioac). in the case of chronic infection, surgical treatment is based on a four stages procedure consisting in surgical sample, surgical excision, bone stabilization and then bone coverage.Antibiotic therapy is systemic and will be started after microbiological samples have been taken. it should be adapted to the microbiological results afterwards. Systemic antibiotic therapy is usually administered initially by the intravenous route, followed by oral, intramuscular, or even subcutaneous administration if possible. The dosage will very often be increased compared to the usual dosages. Drug monitoring is increasingly used. The absence of allergy or drug interaction should also be checked. The patient undergoing antibiotic therapy should be monitored clinically and biologically in a close and prolonged manner because of the frequency of late recurrences.

Building “First Expire, First Out” models to predict food losses at retail due to cold chain disruption in the last mile

Current less-than-truckload (LTL) shipping practices allow for temperature abuse (TA) in the last segment (last mile) of the food supply chain. When this TA is combined with “First In, First Out” product rotation methods, it could lead to food spoilage and food waste; therefore, data-based decision models are needed to aid retail managers. An experiment was designed using pallets (4 layers/pallet × 5 boxes/layer) of commercially produced boneless chicken breast filet trays. The pallets were exposed to 24 h of simulated LTL TA (cyclic 2 h at 4°C, then 2 h at 23 ± 2°C). Filet temperatures were recorded for all 20 boxes using dataloggers with thermocouple wires. Additionally, microbiological sampling of filets [aerobic plate counts (APC) and psychrotrophic plate counts (PSY)] was conducted before (0 h of LTL TA) and after (24 h of LTL TA) the TA experiment for select boxes of the pallet and compared to control filets (maintained at 4°C). After TA, a shelf-life experiment was conducted by storing filets from predetermined boxes at 4°C until spoilage (7 log CFU/ml). Temperature and microbiological data were augmented using Monte Carlo simulations (MC) to build decision making models using two methods; (1) the risk of each box on the pallet reaching the bacterial “danger zone” (&amp;gt;4°C) was determined; and (2) the risk-of-loss (shelf-life &amp;lt; 4 days; minimum shelf-life required to prevent food waste) was determined. Temperature results indicated that boxes on the top and bottom layers reached 4°C faster than boxes comprising the middle layers while the perimeter boxes of each layer reached 4°C faster than centrally located boxes. Shelf-life results indicate simulated LTL TA reduced shelf-life by 2.25 and 1.5 days for APC and PSY, respectively. The first MC method showed the average risk of boxes reaching 4°C after 24 h of simulated LTL TA were 94.96%, 43.20%, 27.20%, and 75.12% for layers 1–4, respectively. The second MC method indicated that exposure at &amp;gt;4°C for 8 h results in a risk-of-loss of 43.8%. The findings indicate that LTL TA decreases shelf-life of chicken breast filets in a heterogenous manner according to location of boxes on the pallet. Therefore, predictive models are needed to make objective decisions so that a “First Expire, First Out” method can be implemented to reduce food wastes due to TA during the last mile.

Investigation of the outbreaks of abortions and orchitis in livestock in Namibia during 2016-2018.

This study investigated outbreaks of seemingly related abortions and orchitis which occurred in the Khomas, Omaheke and Otjozondjupa regions of Namibia from 2016 to 2018, affecting cattle, sheep and goats. Fifty-nine questionnaires were administered, and 48 were completed giving an 81.4% return. The outbreaks were limited to Namibia's east and central regions, mainly on farms rearing cattle, sheep and goats and on farms with a mixture of these species. There was no significant difference between Khomas and other regions on abortion reporting at the farm level [X2(1,N = 48) = 0.0002,p = 0.987851]. However, there was a significant difference in the abortions at the animal level among the three regions [X2(2,N = 6246) = 239.8339,p = .00001]. In addition, the proportions of abortions calculated at the animal level at each farm were significantly different when the Khomas region was compared to the other regions. Seventeen cattle sera, 35 sheep sera, 52 caprine sera, 18 bovine liver samples, one caprine liver, five aborted cattle foetuses, two cattle placentas, 18 testes (one bull, eight bucks and nine rams) and ten bull sheath scrapings were collected and tested. Histopathology, microbiology, serology, immunohistochemistry, real-time PCR and mineral analytical techniques were used to establish the aetiology of the abortion and orchitis outbreaks. The gross and histopathological findings on the 18 testicles were characteristic of chronic orchitis. In aborted foetuses, significant histopathological findings included meconium aspiration, funisitis and cardiomyopathy. Placentitis and endometritis were the primary pathologies observed in cows. The bacteria isolated from microbiological samples included Enterococcus spp. (65.5% [19/29]), Enterobacter spp. (6.9% [2/29]) and Streptococcus spp. (10.3% [3/29]), Trueperella pyogenes (3.4% [1/29]), Stenotrophomonas maltophilia (3.4% [1/29]), Staphylococcus epidermidis (3.4% [1/29]), Providencia rettgeri (3.4% [1/29]) and Acinetobacter lwoffii (3.4% [1/29]), mostly opportunistic bacteria. On mineral analysis, 28%, 33%, 83%, 33% and 17% (n = 18) of cattle livers were low in copper, zinc, manganese, selenium and iron, respectively. Twenty-three percent (12/52) of the caprine sera were positive for Brucella melitensis on the Rose Bengal and complement fixation tests. Thirty-five ovine sera were tested for B. melitensis, B. ovis and Coxiella burnetii, and the prevalence for each was 2.9% (1/35). PCR tests on foetuses were all negative for Brucella spp., Coxiella burnetii, Chlamydia spp., Listeria monocytogenes, Salmonella spp., Campylobacter fetus spp., Leptospira pathogenic strains, bovine viral diarrhoea virus, Rift Valley fever virus, Anaplasma phagocytophilum and bovine herpes virus 4 Campylobacter fetus spp. and Trichomonas foetus spp. The authors concluded that Brucella spp., Enterococcus spp., Escherichia coli, Streptococcus spp., Trueperella pyogenes and Coxiella burnetii could have contributed to this outbreak. Micronutrient imbalances and pathogenic abiotic nanoparticles were also identified as possible contributors to the abortion outbreaks.

Antimicrobial potential of new diclofenac hydrogels for disinfection in regenerative endodontics: An in vitro and ex vivo study.

There is a need to explore new alternatives for root canal disinfection in regenerative endodontics, since the current strategies are far from ideal. Currently, the potential use of diclofenac (DC) is being investigated for controlling root canal infections. The objective was to evaluate the antimicrobial efficacy of novel DC-based hydrogels (DCHs) against polymicrobial biofilms grown in radicular dentine and root canals and to compare results with triantibiotic (TAH) and diantibiotic (DAH) hydrogels, and calcium hydroxide (Ca[OH]2 ). The in vitro antimicrobial activity of intracanal medicaments was evaluated against 3-week-old polymicrobial root canal biofilms grown on human radicular dentine. Dentine samples were obtained and randomly divided into the study groups (n=4/group): (1) 1mg/ml TAH; (2) 1mg/ml DAH; (3) 5% diclofenac (DCH); (4) 2.5% DCH; (5) 1.25% DCH; (6) 1mg/ml DAH + 5% DCH; (7) Ca(OH)2 paste; (8) positive control. The microbial viability, in terms of percentage of intact cell membranes, was assessed after 7 days by confocal scanning laser microscopy (CSLM). The ex vivo efficacy of intracanal medications was evaluated in root canals infected with a polymicrobial suspension. Intracanal microbiological samples at baseline (S1) and 7 days post-treatment (S2) were taken; microbial quantification and cell viability were assessed by quantitative polymerase chain reaction (qPCR) and flow cytometry (FC). The mean Log10 of bacterial DNA copies in root canal samples before (S1) and the Log10 reduction of DNA copies S1-S2 in qPCR were recorded. The absolute value of total cells stained, and the percentage reduction of intact membrane cells after treatment (S1-S2), were analysed by FC. Global comparison was done using the Kruskal-Wallis test, whilst the Mann-Whitney U test was used for pair-by-pair comparison. Confocal scanning laser microscopy analysis indicated that the greatest effectiveness was obtained with 5% DCH, showing significant differences with respect to the other groups (p < .001). In root canals, the highest Log10 DNA reduction S1-S2 was obtained with 5% DCH and TAH, with no differences between them. The results of FC showed that only 5% DCH proved significantly superior to the other treatments. Sodium DC hydrogels demonstrate antimicrobial efficacy against endodontic biofilms.

Antibiotic Stewardship (ABS) im Kontext schwerer Infektionen

HintergrundSchwere Infektionen stellen aufgrund der häufigen Komplikationen und hohen Sterblichkeit eine große klinische Herausforderung dar.Ziel der Arbeit (Fragestellung)Am Beispiel der Sepsis, gramnegativer Blutstrominfektionen und COVID-19 sollen Anwendung und Bedeutung von Antibiotic Stewardship(ABS)-Strategien im Kontext schwerer Infektionen erläutert werden.Material und MethodeEs erfolgen die Zusammenfassung der aktuellen Literatur und Empfehlungen mit Fokus auf klinische ABS-Implikationen bei schweren Infektionen wie Sepsis, gramnegativen Blutstrominfektionen und COVID-19.ErgebnisseDie aktualisierte Sepsis-Leitlinie schlägt ein differenziertes Vorgehen zum Beginn der Antibiotikatherapie bei Patient*innen mit Sepsis, aber ohne Schock vor. Bei unsicherer Diagnose und bei fehlendem Schock kann der Zeitraum bis zur Einleitung der Antibiotikatherapie auf 3 h ausgedehnt werden. In dieser Zeit gilt es, die Differenzialdiagnostik und ein adäquates mikrobiologisches Sampling zu komplettieren, um eine Antibiotikatherapie aufgrund belastbarer Befunde zu initiieren oder alternativ zu verwerfen. Bei unkomplizierten gramnegativen Blutstrominfektionen ist eine 7‑tägige antibiotische Therapie ausreichend. Eine regelhafte Kombinationstherapie bei Blutstrominfektionen mit Pseudomonas aeruginosa ist aufgrund der bisher vorliegenden Daten nicht erforderlich. Bakterielle Koinfektionen bei COVID-19 sind selten. Eine prophylaktische antibiotische Therapie ist daher nicht gerechtfertigt. Bakterielle Sekundärinfektionen (> 48 h nach Hospitalisierung) treten v. a. bei kritisch kranken COVID-19-Patient*innen auf. Bei pulmonalen Ko- oder Sekundärinfektion ist ein therapeutisches Vorgehen entsprechend den Pneumonie-Leitlinien geboten.SchlussfolgerungenTrotz der Schwere und Komplexität der genannten Infektionen gelten die grundlegenden Prinzipien des Antibiotic Stewardship (ABS) im Sinne einer rationalen Antiinfektivaverordnung hinsichtlich Substanz, Dosis und Dauer der Therapie.

Antibiotic treatment duration in diverticulitis, complicated urinary tract infection, and endocarditis: a retrospective, single-center study

ObjectivesDespite the availability of international guidelines advocating shorter treatment durations, nonadherence to them is common. We assessed duration of antibiotic treatment for diverticulitis, complicated urinary tract infection (UTI), and endocarditis. MethodsMedical records of patients hospitalized with the previously stated diseases in 2017 and 2018 were randomly selected at a Swiss tertiary care hospital. The appropriateness of antibiotic treatment duration was assessed according to international and local guidelines. ResultsA total of 243 patients were included in the study: 100 with diverticulitis, 200 with complicated UTI, and 43 with endocarditis. The dherence to local and international guidelines was 11% and 18% in diverticulitis, 39% and 40% in complicated UTI, and 84% and 86% in endocarditis, respectively. Nonadherence was primarily due to the prolonged treatment in diverticulitis and complicated UTI with a median duration of antibiotic treatment of 11 days (interquartile range 10-13) and 14 days (interquartile range 10-15), respectively. When pooling diverticulitis and complicated UTI cases, the identification of a pathogen in any microbiological sample was associated with an improved adherence to local guidelines in addition to hospitalization in a medical ward and infectious diseases consultation. ConclusionProlonged courses of antibiotic treatment were common and the treatment adherence to guidelines were poor in diverticulitis, moderate in complicated UTI, and excellent in endocarditis.

Borescope examination and microbial culture results of endoscopes in a tertiary care hospital led to changes in storage protocols to improve patient safety

Flexible endoscopes are highly versatile and useful medical instruments, and their proper reprocessing is critical to patient health and safety. The value of routine visual inspections and surveillance of endoscopes in a tertiary care hospital was assessed by performing borescope examinations and microbial sampling on respiratory, gastro-intestinal (GI), and urological endoscopes. A total of 42 endoscopes were cultured, and 36 endoscopes were examined with a borescope. The flush-brush-flush method was used to culture the endoscopes. Microbiologic sampling water was vortexed prior to being suctioned through a membrane filter device. The membrane was plated on a blood agar plate and incubated at 37°C for 48 hours. A 1.9-meter borescope with a 1.6mm diameter was used to perform borescope examinations in an antegrade and retrograde approach. Positive microbial cultures were seen in 28% of bronchoscopes, 22% of GI endoscopes, and 30% of urological endoscopes. Only Gram-positive colonies were identified in endoscopes that had microbial growth. Borescope examinations revealed multiple abnormalities and damage including channel shredding, filamentous debris, water retention, discoloration, dents, and red particles. Borescope examination and microbial sampling should be used routinely to assure endoscopic safety. Borescope examination enabled us to visualize structural damage, foreign material, and moisture within endoscopes. The structural damages and the particles found in endoscopes resulted in timely repair and discontinuation of this type of distal end protectors in our facility.

Patients with obstructive sleep apnea can favor the predisposing factors of periodontitis by the presence of P. melaninogenica and C. albicans, increasing the severity of the periodontal disease.

ObjectiveThe aim of this study was to analyze the cultivable oral microbiota of patients with obstructive sleep apnea (OSA) and its association with the periodontal condition.MethodsThe epidemiology profile of patients and their clinical oral characteristics were determined. The microbiota was collected from saliva, subgingival plaque, and gingival sulcus of 93 patients classified into four groups according to the periodontal and clinical diagnosis: Group 1 (n = 25), healthy patients; Group 2 (n = 17), patients with periodontitis and without OSA; Group 3 (n = 19), patients with OSA and without periodontitis; and Group 4 (n = 32), patients with periodontitis and OSA. Microbiological samples were cultured, classified, characterized macroscopically and microscopically, and identified by MALDI-TOF-MS. The distribution of complexes and categories of microorganisms and correlations were established for inter- and intra-group of patients and statistically evaluated using the Spearman r test (p-value <0.5) and a multidimensional grouping analysis.ResultThere was no evidence between the severity of OSA and periodontitis (p = 0.2813). However, there is a relationship between the stage of periodontitis and OSA (p = 0.0157), with stage III periodontitis being the one with the highest presence in patients with severe OSA (prevalence of 75%; p = 0.0157), with more cases in men. The greatest distribution of the complexes and categories was found in oral samples of patients with periodontitis and OSA (Group 4 P-OSA); even Candida spp. were more prevalent in these patients. Periodontitis and OSA are associated with comorbidities and oral conditions, and the microorganisms of the orange and red complexes participate in this association. The formation of the dysbiotic biofilm was mainly related to the presence of these complexes in association with Candida spp.ConclusionPeriodontopathogenic bacteria of the orange complex, such as Prevotella melaninogenica, and the yeast Candida albicans, altered the cultivable oral microbiota of patients with periodontitis and OSA in terms of diversity, possibly increasing the severity of periodontal disease. The link between yeasts and periodontopathogenic bacteria could help explain why people with severe OSA have such a high risk of stage III periodontitis. Antimicrobial approaches for treating periodontitis in individuals with OSA could be investigated in vitro using polymicrobial biofilms, according to our findings.

Standardization of Epidemiological Surveillance of Group A Streptococcal Cellulitis.

Cellulitis is an acute bacterial infection of the dermis and subcutaneous tissue usually found complicating a wound, ulcer, or dermatosis. This article provides guidelines for the surveillance of cellulitis. The primary objectives of cellulitis surveillance are to (1) monitor trends in rates of infection, (2) describe the demographic and clinical characteristics of patients with cellulitis, (3) estimate the frequency of complications, and (4) describe the risk factors associated with primary and recurrent cellulitis. This article includes case definitions for clinical cellulitis and group A streptococcal cellulitis, based on clinical and laboratory evidence, and case classifications for an initial and recurrent case. It is expected that surveillance for cellulitis will be for all-cause cellulitis, rather than specifically for Strep A cellulitis. Considerations of the type of surveillance are also presented, including identification of data sources and surveillance type. Minimal surveillance necessary for cellulitis is facility-based, passive surveillance. Prospective, active, facility-based surveillance is recommended for estimates of pathogen-specific cellulitis burden. Participant eligibility, surveillance population, and additional surveillance considerations such as active follow-up of cases, the use of International Classification of Disease diagnosis codes, and microbiological sampling of cases are discussed. Finally, the core data elements to be collected on case report forms are presented.

Cytocompatibility and antimicrobial activity of a novel endodontic irrigant combining citric acid and chlorhexidine

ObjectivesThis study aimed to evaluate the antibacterial ability and cytocompatibility of a new irrigant solution for endodontic treatment composed of 10% citric acid (CA) and 1% chlorhexidine (CHX). MethodsThirty-five extracted single-canal human teeth were selected and de-crowned. Canal systems (n = 7/group) were infected with Enterococcus faecalis for 4 weeks and subject to irrigation with 1% CHX; 10% CA; irrigating solution 10% CA associated with 1% CHX (CACHX); 2.5% NaOCl or sterile water (control). Microbiological samples were collected immediately and 18 h after irrigation (enriched samples). The canals were filled with culture medium post irrigation to verify the bacterial presence/absence qualitatively and quantitatively through colony counting (log10 CFU/mL). A multiparametric assay was performed after exposure of human periodontal ligament fibroblasts (HPdLF) to the test solutions. The Kruskal-Wallis test with Dunn´s post-test and Fisher's exact test were employed at the 95% confidence level to compare differences among groups. ResultsAll tested solutions were cytocompatible with human periodontal ligament fibroblasts. No difference was observed on antibacterial activity between 1% CHX, 10% CA, CACHX and 2.5% NaOCl (p > 0.05). Eighteen hours after irrigation, CACHX samples were the only that did not present E. faecalis in the root canal system. ConclusionsThe demonstrated good in vitro biocompatibility and elimination of E. faecalis suggest a potential use of 10% CA associated with 1% CHX as a solution for microbiological control during endodontic treatment. Clinical RelevanceIrrigants play an essential role during endodontic therapy. This irrigating solution, based on the association of 10% citric acid with 1% chlorhexidine, seems viable for clinical procedures.