Airborne SARS-CoV-2 RNA excretion by patients with COVID-19 on different oxygen-delivery systems: a prospective observational study.

BACKGROUND: Concerns persist regarding the risk of airborne SARS-CoV-2 transmission by patients with COVID-19 on various modalities of oxygen therapy, such as high-flow nasal cannula (HFNC). AIM: We aimed to compare the presence of airborne RNA in air samples between groups of patients with COVID-19 on different oxygen-delivery systems. We also explored factors that were associated with SARS-CoV-2 RNA positivity in air samples. RESULTS: Air samples were positive for SARS-CoV-2 RNA in three of 39 patients (8%) on HFNC, 0 of 13 (0%) on masks, versus five of 20 (25%) on nasal cannula. Odds ratio for air sample positivity was 0.52 (95% confidence interval (CI) 0.11-2.34) when comparing HFNC vs non-HFNC group, and 5.78 (1.24-27.01) for nasal cannula vs non-nasal cannula group. Patients with positive air samples in comparison with those with negative air samples were sampled earlier after symptoms onset (median: 7 vs 10 days; P=0.04) and had lower Ct values of diagnostic nasopharyngeal samples (median: 22 vs 26; P=0.02). CONCLUSIONS: Air sample positivity was not related to oxygen support device but to viral load. These data suggest that the use of personal protection equipment should be based on risk management according to viral load rather than oxygen support device.

Airborne SARS-CoV-2 in home and hospital environments investigated with a high-powered air sampler.

BACKGROUND: The initial aim was to study the effects of face masks worn by recently infected individuals on the airborne spread of SARS-CoV-2, but findings motivated us to proceed with comparing the presence of SARS-CoV-2 in air samples near infected individuals at home with those near infected intensive care unit (ICU) patients. AIM: To assess the presence of SARS-CoV-2 in the air of homes of infected individuals and in ICU rooms of critically ill patients with COVID-19 who were undergoing different forms of potential aerosol-generating medical procedures. METHODS: A high-volume air sampler method was developed that used a household vacuum cleaner with surgical face masks serving as sample filters. SARS-CoV-2 RNA was harvested from these filters and analysed by polymerase chain reaction. Fog experiments were performed to visualize the airflow around the air sampler. Air samples were acquired in close proximity of infected individuals, with or without wearing face masks, in their homes. Environmental air samples remote from these infected individuals were also obtained, plus samples near patients in the ICU undergoing potential aerosol-generating medical procedures. FINDINGS: Wearing a face mask resulted in a delayed and reduced flow of the fog into the air sampler. Face masks worn by infected individuals were found to contain SARS-CoV-2 RNA in 71% of cases. SARS-CoV-2 was detected in air samples regardless of mask experiments. The proportion of positive air samples was higher in the homes (29/41; 70.7%) than in the ICU (4/17; 23.5%) (P < 0.01). CONCLUSION: SARS-CoV-2 RNA could be detected in air samples by using a vacuum cleaner based air sampler method. Air samples in the home environment of recently infected individuals contained SARS-CoV-2 RNA nearly three times more frequently by comparison with those obtained in ICU rooms during potential aerosol-generating medical procedures.

Transmission of SARS-CoV-2 among healthcare workers and patients in a teaching hospital in the Netherlands confirmed by whole-genome sequencing.

AIM: To investigate the sources of infection among healthcare workers (HCWs) and patients in a teaching hospital in the Netherlands during the early stages of the coronavirus disease 2019 (COVID-19) pandemic using epidemiological and whole-genome sequencing data. METHODS: From 3rd April to 11th May 2020, 88 HCWs and 215 patients were diagnosed with COVID-19. Whole-genome sequences were obtained for 30 HCWs and 20 patients. RESULTS: Seven and 11 sequence types were identified in HCWs and patients, respectively. Cluster A was the most common sequence type, detected in 23 (77%) HCWs; of these, 14 (61%) had direct patient contact and nine (39%) had indirect patient contact. In addition, seven patients who were not hospitalized in the COVID-19 cohort isolation ward who became positive during their admission were infected with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) cluster A. Following universal masking of all HCWs and emphasis on physical distancing during meals and breaks, no further evidence was found for patient-to-HCW or HCW-to-HCW transmission or vice versa. CONCLUSION: The finding that patients and HCWs were infected with SARS-CoV-2 cluster A suggests both HCW-to-HCW and HCW-to-patient transmission.

Is there an adequate alternative to commercially manufactured face masks? A comparison of various materials and forms.

BACKGROUND: There is a worldwide shortage of medical-grade face masks. Donning masks can play an important role in curbing the spread of SARS-CoV-2. AIM: To conclude whether there is an effective mask for the population to wear in public that could easily be made during a medical face mask shortage using readily available materials. METHODS: We determined the effectiveness of readily available materials and models for making a face mask. The outcomes were compared with N95/FFP2/KN95 masks that entered the Netherlands in April-May 2020. Masks were tested to determine whether they filtered a minimum of 35% of 0.3-µm particles, are hydrophobic, seal on the face, are breathable, and can be washed. FINDINGS: Fourteen of the 25 (combinations of) materials filtered at least 35% of 0.3-µm particles. Four of the materials proved hydrophobic, all commercially manufactured filters. Two models sealed the face. Twenty-two of the 25 materials were breathable at <0.7 mbar. None of the hydrophobic materials stayed intact after washing. CONCLUSIONS: It would be possible to reduce the reproduction rate of SARS-CoV-2 from 2.4 to below one if 39% of the population would wear a mask made from ePM1 85% commercially manufactured filter fabric and in a duckbill form. This mask performs better than 80% of the imported N95/FFP2/KN95 masks and provides a better fit than a surgical mask. Two layers of quilt fabric with a household paper towel as filter is also a viable choice for protecting the user and the environment.

Epstein-Barr virus mimicking lymphoma--a case report.

A 50-year-old male without a relevant medical history came to the emergency department with fever, muscle pain and fatigue without any localising symptoms. Blood and urine cultures remained negative. Laboratory work- up showed elevated liver enzymes and lactate dehydrogenase; Epstein-Barr virus (EBV) serology was negative. Additional imaging showed a splenomegaly and cervical, axillary, mediastinal and hilar lymphadenopathy. Pathological examination of one of the lymph nodes and bone marrow biopsy revealed a peripheral T-cell non-Hodgkin's lymphoma not otherwise specified. Before the start of treatment he was asymptomatic, the laboratory results had normalised and the EBV polymerase chain reaction was strongly positive. Computed tomography scan was repeated and showed complete remission of the lymphadenopathy and normalised spleen volume. Follow-up bone marrow analysis including clonal rearrangement of the T-cell receptor after three months and one year revealed a decreasing clonal T-cell population (41%, 39% and 11% respectively). In conclusion, this was an extreme course of an EBV infection. The clinical relevance of the remaining small monoclonal T-cell population detectable in the bone marrow is unclear.

Comparison of the COBAS AMPLICOR MTB and BDProbeTec ET assays for detection of Mycobacterium tuberculosis in respiratory specimens.

The performances of the BDProbeTec ET (Becton Dickinson) and COBAS AMPLICOR MTB (Roche) were retrospectively evaluated for detecting Mycobacterium tuberculosis complex in various respiratory specimens. The BACTEC and MGIT liquid culture system (Becton Dickinson) was used as a reference method. A total of 824 respiratory specimens, comprised of sputa, bronchoalveolar lavage fluid, and bronchial and tracheal aspirates from 580 patients, were evaluated. Out of 824 clinical specimens, 109 specimens from 43 patients were culture positive for M. tuberculosis. Of these 109 specimens, 67 were smear positive, 85 were positive by the COBAS AMPLICOR MTB test, and 94 were positive by the BDProbeTec ET. Of the 715 culture-negative specimens, 17 were positive by the auramine staining, 11 were positive by the COBAS AMPLICOR MTB test, and 12 were positive by the BDProbeTec ET. After discrepancy analysis and review of the patients' clinical data, 130 specimens from 50 patients were considered "true-positive" specimens. This resulted in the following sensitivities: microscopy, 61.5%; COBAS AMPLICOR MTB test, 78.0%; and BDProbeTec ET, 86.2%. The specificities of each system, based on the clinical diagnosis, were 99.7% for microscopy, 99.9% for the COBAS AMPLICOR MTB test, and 99.9% for the BDProbeTec ET. The data presented represent a considerable number of specimens evaluated with a considerable number of culture- and auramine-positive and culture-positive and auramine-negative results and therefore give a realistic view of how the data should be interpreted in a daily routine situation. Specifically, the data with regard to the culture-positive and auramine-negative specimens are useful, because in a routine situation, auramine-negative specimens are sometimes accepted, on clinical indications, to be analyzed by an amplification method.

Enterobacter species in a pediatric hospital: horizontal transfer or selection in individual patients?

Enterobacter species were studied longitudinally in a children's hospital. In total, 287 Enterobacter isolates were obtained from 171 children in 15 different wards (from March 1995 through April 1997). Strains were typed by random amplified polymorphic DNA and pulsed-field gel electrophoresis, which were concordant in outcome. In total, 97 DNA types and 199 colonization events were identified. A predominant clone was isolated 111 times from 62 children; another clone was isolated 19 times from 10 patients. These clones caused 36% of all colonizations. In 34% of the children, Enterobacter clones were found in 2-4 patients. The remaining colonizations were due to unique Enterobacter isolates. A large proportion of the Enterobacter strains was acquired through cross-transmission. This finding contrasts with the prevailing opinion that resistant Enterobacter strains are selected primarily from the patient's own gut flora.

Improved detection of methicillin-resistant Staphylococcus aureus using phenyl mannitol broth containing aztreonam and ceftizoxime.

We tested a phenyl mannitol broth containing ceftizoxime and aztreonam (PHMB(+)) for detection of methicillin-resistant Staphylococcus aureus (MRSA) with reference MRSA strains and, subsequently, with clinical samples (n = 1,098). All reference MRSA strains induced color change in PHMB(+) after 24 to 72 h of incubation. In a clinical setting, 40 MRSA strains were detected with PHMB(+), compared with only 23 detected with a routine method. Thus, this selective broth significantly (P < 0.001) improved the rate of MRSA detection.

Evaluation of a new commercial immunoassay for rapid detection of Campylobacter jejuni in stool samples.

In order to evaluate a new commercial enzyme immunoassay (ProspecT Campylobacter Microplate Assay; Alexon-Trend, USA) for the detection of Campylobacter jejuni and Campylobacter coli in stool samples, 30 faecal specimens known to be culture-positive for Campylobacter jejuni were tested with the new assay. The detection limit was approximately 3 x 10(6)/ml in faecal suspensions. The sensitivity relative to culture was 80% (24/30). All of the 24 positive samples, except for one, remained positive after being stored at -20 degrees C for 60 days. The specificity of the test was 100%. Interestingly, 6 of 11 additional Campylobacter jejuni culture-positive samples that had been obtained from patients with Guillain-Barré syndrome and stored at -20 degrees C for periods of up to 5 years tested positive in the assay. The performance of the assay indicates that it has potential value for use in future early intervention studies.

Role of bacterial virulence factors and host factors in the outcome of Escherichia coli bacteraemia.

In a study of the role of virulence factors in the outcome of Escherichia coli bacteraemia, blood isolates from 30 hospitalised patients were characterised with regard to O and K antigens, P and type 1 fimbriae, haemolysin production, cytonecrotising factor 1 production, serum resistance, ability to activate neutrophils and resistance to killing. Patients were analysed to identify host factors contributing to morbidity and mortality. In univariate analyses the presence of a K antigen, type 1 fimbriae, absence of haemolysin production, serum resistance and resistance to killing were associated with morbidity and mortality. In multivariate analyses only the absence of haemolysin production was associated with morbidity and mortality, after taking host factors into account. These preliminary findings suggest that host factors override bacterial virulence factors in determining the course of Escherichia coli bacteraemia. The negative association between haemolysin production and clinical deterioration during Escherichia coli bacteraemia might indicate predominance of less virulent strains in patients with other risk factors for morbidity and mortality or inactivation of neutrophil products needed for host defence.

An antibiotic policy to prevent emergence of resistant bacilli.

BACKGROUND: Fear of infection in neonatal intensive care units (NICUs) often leads to early use of empiric broad-spectrum antibiotics, a strategy that selects for resistant bacteria. We investigated whether the emergence of resistant strains could be halted by modifying the empiric antibiotic regimens to remove the selective pressure that favours resistant bacteria. METHODS: Two identical NICUs were assigned to different empiric antibiotic regimens. On unit A, penicillin G and tobramycin were used for early-onset septicaemia, flucloxacillin and tobramycin were used for late-onset septicaemia, and no broad-spectrum beta-lactam antibiotics, such as amoxicillin and cefotaxime were used. In unit B, intravenous amoxicillin with cefotaxime was the empiric therapy. After 6 months of the study the units exchanged regimens. Rectal and respiratory cultures were taken on a weekly basis. FINDINGS: There were 436 admissions, divided equally between the two regimens (218 in each). Three neonates treated with the penicillin-tobramycin regimen became colonised with bacilli resistant to the empirical therapy used versus 41 neonates on the amoxicillin-cefotaxime regimen (p<.0001). The relative risk for colonisation with strains resistant to the empirical therapy per 1000 patient days at risk was 18 times higher for the amoxicillin-cefotaxime regimen compared with the penicillin-tobramycin regimen (95% CI 5.6-58.0). Enterobacter cloacae was the predominant bacillus in neonates on the amoxicillin-cefotaxime regimen, whereas Escherichia coli predominated in neonates on the penicillin-tobramycin regimen. These colonisation patterns were also seen when the units exchanged regimens. INTERPRETATION: Policies regarding the empiric use of antibiotics do matter in the control of antimicrobial resistance. A regimen avoiding amoxicillin and cefotaxime restricts the resistance problem.

Persistence of clones of coagulase-negative staphylococci among premature neonates in neonatal intensive care units: two-center study of bacterial genotyping and patient risk factors.

From 1 January 1995 until 1 January 1996, we studied the molecular epidemiology of blood isolates of coagulase-negative staphylococci (CoNS) in the Neonatal Intensive Care Units (NICUs) of the Sophia Children's Hospital (SCH; Rotterdam, The Netherlands) and the Wilhelmina Children's Hospital (WCH; Utrecht, The Netherlands). The main goal of the present study was to detect putatively endemic clones of CoNS persisting in these NICUs. Pulsed-field gel electrophoresis was used to detect the possible presence of endemic clones of clinical significance. In addition, clinical data of patients in the SCH were analyzed retrospectively to identify risk factors for the acquisition of positive blood cultures. In both centers, endemic CoNS clones were persistently present. Thirty-three percent of the bacterial isolates derived from blood cultures in the SCH belonged to a single genotype. In the WCH, 45% of all bacterial strains belonged to a single clone. These clones were clearly different from each other, which implies that site specificity is involved. Interestingly, we observe that the clonal type in the SCH differed significantly from the incidentally occurring strains with respect to both the average pH and partial CO2 pressure of the patient's blood at the time of bacterial culture. We found that the use of intravascular catheters, low gestational age, and a long hospital stay were important risk factors for the development of a putative CoNS infection. When the antibiotic susceptibility of the bacterial isolates was assessed, a clear correlation between the nature of the antibiotics most frequently used as a first line of defense versus the resistance profile was observed. We conclude that the intensive use of antibiotics in an NICU setting with highly susceptible patients causes selection of multiresistant clones of CoNS which subsequently become endemic.

Susceptibility of lipopolysaccharide-responsive and -hyporesponsive ItyS Mice to infection with rough mutants of Salmonella typhimurium.

The R5 (chemotype Rb) but not the R10 (chemotype Rd) mutant of murine pathogen Salmonella typhimurium 395MS was extremely virulent in intraperitoneal infections of C57BL/10ScCr mice carrying the ityS and lpsD alleles. C57BL/6J (ityS lpsN) and C3H/HeJ (ityR lpsD) mice showed a much higher resistance to the R5 mutant. Further studies were performed with peritoneal macrophages in vitro in order to elucidate susceptibility in lipopolysaccharide (LPS)-hyporesponsive mice carrying ItyS. The intracellular killing capacity of the ItyS LpsD macrophages was lower than that of the ItyS LpsN macrophages for the R5 mutant and may partly explain the increased susceptibility of the ItyS LpsD mice. The deep rough mutant, R10, was rapidly killed intracellularly by the ItyS LpsD macrophages. Processing of the bacteria in macrophages that had phagocytosed R5 or R10 bacteria was followed for up to 18 days by endotoxin measurements (limulus assay) and immunostaining, with monoclonal antibodies to various parts of the LPS molecule being used. Only 0.1% or less of the macrophage-associated bacteria remained alive after 48 h of incubation, and none were alive on day 7. Although immunostaining showed that LPS was present in both the LpsD and LpsN macrophages during the whole incubation period of 18 days, endotoxin activity in the LpsD macrophages on day 7 was lower than that in the LpsN macrophages, indicating that qualitative modifications of the chemical composition or physical state of the LPS molecule occurred. The interleukin-6 response in the ItyS LpsD macrophages was delayed and of shorter duration compared with that in the ItyS LpsN macrophages. The results suggest that the difference between the LPS-hyporesponsive and -responsive ItyS mice in susceptibility to infection with the R5 mutant was due to the lower activation state of the LpsD macrophages during infection, leading to a lower intracellular bactericidal systems of the macrophages. A rapid killing of the bacterium should restrict the infection and may partly compensate for a diminished inflammatory response. The persistence of LPS within the cells is discussed.

Lack of association between hemolysin production and acute inflammation in human urinary tract infection.

Hemolysins are cytolytic proteins which have been extensively characterized at the molecular level, however, their in vivo functions remain unclear. This study analyzed the association of hemolysin production with the inflammatory response in patients with urinary tract infection (UTI). Infants and children with their first episode of UTI (n = 644) were followed prospectively. The body temperature, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), urinary leucocyte count and renal concentrating capacity were used as measures of the inflammatory response. The hemolytic genotype (hly) of the Escherichia coli strain from each UTI episode was defined by DNA-DNA hybridization, and the phenotype by hemolysis in blood agar. There was no significant increase in the level of fever, CRP, ESR, or decrease in renal concentrating capacity during UTI episodes caused by hly positive compared to hly negative E. coli. Multiple regression analysis did not demonstrate significant associations of hly with elevated fever, CRP, ESR or reduced renal concentrating capacity. In contrast, patients infected with P fimbriated E. coli strains had higher fever, CRP, ESR and lower renal concentrating capacity than those infected with other strains. This association was not influenced by the hly genotype of the P fimbriated strains. The frequency of hly+ strains was not significantly higher in the subset of patients assigned a diagnosis of acute pyelonephritis compared to asymptomatic bacteriuria. This was in contrast to P fimbriae, which were accumulated in acute pyelonephritis. The results suggested that the acute inflammatory response to E. coli UTI is independent of hemolysin production. The inflammatogenic potential of uropathogenic E. coli clones was better described by the presence or absence of P-fimbriae than by hemolysin.

Blood group non-secretors have an increased inflammatory response to urinary tract infection.

The aim of this study was to assess the possible relationship between secretor state and the inflammatory response to urinary tract infection (UTI). Girls with recurrent UTI were prospectively studied. They included 61 secretor and 23 non-secretor individuals with 604 episodes of recurrent UTI. The response to each UTI episode was measured as the levels of C-reactive protein, erythrocyte sedimentation rate and the body temperature as well as renal concentrating capacity and pyuria. The levels of C-reactive protein, erythrocyte sedimentation rate and the body temperature were significantly higher in non-secretors than in secretors (p less than 0.04). As a consequence, non-secretors had an increased probability of being assigned a diagnosis of acute pyelonephritis rather than asymptomatic bacteriuria (p less than 0.05). The higher inflammatory response in non-secretors was independent of the Gal alpha 1-4Gal beta adhesin expression of the infecting Escherichia coli strains. The increased inflammatory response to UTI in non-secretors might explain the accumulation of these individuals among patients with renal scarring.

Resident colonic Escherichia coli strains frequently display uropathogenic characteristics.

Bacterial factors associated with long-term persistence in the colon have not been defined. Individual Escherichia coli strains in the colonic flora of 13 schoolgirls with asymptomatic bacteriuria were identified by electromorphic typing of chromosomally encoded enzymes and defined as resident or transient. The strains were characterized as to serotype, receptor specificity, and adherence to the human colonic epithelial cell line HT-29. Colonic resident strains expressed P fimbriae, adhered to colonic epithelial cells via a mannose-resistant mechanism, and expressed the uropathogenic serotypes O1, O2, O6, O7, O18, O25, or O75 more often than did the transient strains, which were often nontypeable. The serotype and hemagglutination pattern were generally retained during intestinal carriage, in contrast to the loss of such properties upon prolonged colonization of the urinary tract. P fimbriae with Gal alpha 1----4Gal beta-specific adherence may, in fact, have evolved to increase persistence in the colon.

Intestinal colonization with Enterobacteriaceae in Pakistani and Swedish hospital-delivered infants.

Rectal cultures from Swedish and Pakistani hospital-delivered newborn infants were analysed regarding the early acquisition of enterobacteria. Swedish infants were delivered vaginally, Pakistani infants were delivered either vaginally or by caesarean section. The Swedish infants were all breast-fed, whereas breastfeeding was incomplete and often started late among the Pakistani infants. Both groups of Pakistani infants were more rapidly colonized with enterobacteria than were the Swedish infants. Cultures from Swedish infants seldom yielded more than one kind of enterobacteria; E. coli and Klebsiella were most frequently isolated. E. coli dominated in both Pakistani groups, but especially caesarean section delivered infants were in addition often colonized with Proteus, Klebsiella, Enterobacter or Citrobacter species. Breastfeeding from the first day of life reduced colonization with Klebsiella/Enterobacter/Citrobacter. The results suggest that environmental exposure, delivery mode and early feeding habits all influence the early intestinal colonization with enterobacteria.