Nosocomial outbreak of monoclonal VIM carbapenemase-producing Enterobacter cloacae complex in an intensive care unit during the COVID-19 pandemic: an integrated approach.

BACKGROUND: An outbreak of VIM carbapenemase-expressing Enterobacter cloacae complex occurred between March and October 2020 in an intensive care unit (ICU) of a tertiary care and teaching hospital in France. At the same time, the hospital was facing the COVID-19 first wave. AIM: To describe the management of an outbreak caused by a VIM-producing Enterobacter cloacae complex strain during the COVID-19 pandemic in an ICU and to show the importance of an integrated approach. METHODS: A multi-focal investigation was conducted including descriptive and molecular epidemiology, environmental screening, and assessment of infection prevention and control measures. FINDINGS: A total of 14 cases were identified in this outbreak with a high attributable mortality rate (85.7%). The outbreak management was coordinated by a crisis cell, and involved the implementation of multi-disciplinary actions such as: enhanced hygiene measures, microbiological and molecular analysis of patients and environmental E. cloacae complex strains, and simulation-based teaching. All 23 E. cloacae complex strains isolated from patients and environment samples belonged to multi-locus sequence type ST78 and carried bla-VIM4 gene. Using Fourier transform infrared spectroscopy, all but two isolates were also found to belong to a single cluster. Although the source of this outbreak could not be pinpointed, the spread of the strain was controlled thanks to this multi-focal approach and multi-disciplinary implementation. CONCLUSION: This investigation highlighted the usefulness of Fourier transform infra-red spectroscopy in the rapid typing of outbreak strains as well as the importance of an integrated approach to successfully fight against multidrug-resistant micro-organism dissemination and healthcare-associated infections.

Characterization of a multidrug-resistant Klebsiella pneumoniae ST607-K25 clone responsible for a nosocomial outbreak in a neonatal intensive care unit.

PURPOSE: Multidrug-resistant Klebsiella pneumoniae strains are regularly involved in hospital outbreaks. This study describes an ESBL-producing K. pneumoniae clone (ST607-K25) responsible for a nosocomial outbreak in a neonatal intensive care unit. METHODOLOGY: Fourteen strains isolated from 13 patients were included. Antimicrobial susceptibility testing was performed by the agar diffusion method. A clonal link was first investigated by fingerprinting (ERIC-PCR and REP-PCR) then confirmed by MLST. Characterization was performed by molecular detection and identification of several drug resistance and virulence determinants. RESULTS: All strains expressed the same antibiotype, combining ESBL production, fluoroquinolones and aminoglycoside resistance, except for one which remained susceptible to fluoroquinolones. Fingerprinting methods confirmed the clonal link and MLST identified a ST607 clone. Molecular investigations revealed: (I) genes encoding for two narrow-spectrum beta-lactamases (SHV-1 and TEM-1) and an ESBL (CTX-M-15); (II) absence of any chromosomal mutation in quinolone resistance-determining- regions (QRDR) of gyrA/gyrB and parC/parE genes; (III) genes encoding for three plasmid-mediated quinolone-resistance (PMQR) determinants: oqxAB (14/14), aac(6')-Ib-cr (14/14) and qnrB (13/14); (IV) production of a K25 capsule; and (V) carriage of three genes encoding for virulence factors: mrkD (type 3 fimbriae) (14/14), ybts (yersiniabactin) (12/14) and entB (enterobactin) (14/14). CONCLUSION: We described a multidrug-resistant Kp ST607 clone responsible for a nosocomial outbreak in vulnerable and premature newborns. Molecular investigations allowed us to identify several resistance factors responsible for ESBL production (CTX-M-15) and quinolone resistance (three PMQR determinants). The detection of a gene (ybtS) belonging to the high-pathogenicity island yersiniabactin could partly explain its high colonization and diffusion potential.

Breastmilk donations: Bacteriological assessment, analysis of causes of non-compliance and suggestions for improvement.

A total of 1099 breastmilk donations received by the milk bank at the Amiens University Hospital from January to June 2016 were assessed for bacteriological contamination according to French regulations. This consisted in enumerating the total aerobic flora before and after heat treatment as well as the specific enumeration of coagulase-positive staphylococci. Results above the mandatory limits for at least one of these parameters were found in 25.9% of the donations, resulting in the destruction of approximately one-quarter of the volume of the donations (∼195L). This is a huge loss in both economic and health-related terms for neonates, especially for pre-terms. To identify ways to improve the bacteriological assessment results and reduce the percentage of discarded milk, an analysis of the causes was conducted. The two main causes of non-compliance were the detection of a cultivable aerobic flora after heat treatment and the presence of coagulase-positive staphylococci above the mandatory limit (11.7% and 11.2% of the tested donations, respectively). Bacillus spp. were the leading cause of post-heat-treatment non-compliance. Therefore, the implementation of better environmental control could help reduce this kind of contamination. As for samples harboring coagulase-positive staphylococci, a further detection of toxins using molecular biology techniques could help discriminate actual health-hazardous donations that have to be destroyed while enabling the use of toxin-negative donations. Nevertheless, the economic viability of this proposal needs to be further assessed because these techniques are costly. Finally, a change in breastmilk dilutions used to enumerate the total aerobic flora to better reflect the actual level of these bacteria in the milk was proposed. Indeed, the comparison of various combinations of milk dilutions led to the conclusion that the association of the 1/10 and 1/100 dilutions was the best compromise between technical ease of enumeration and ensuring the safety of the donations. Implementing these suggestions would help reduce the rate of non-compliance and give better access to safe breastmilk donations for neonates.

[Quantitative assessment of fungal risk in the case of construction works in healthcare establishments: Proposed indicators for the determination of the impact of management precautions on the risk of fungal infection]. / Appréciation quantitative du risque fongique en cas de travaux en établissements de santé : propositions d'indicateurs d'impact des mesures de gestion du risque infectieux fongique.

Construction works in healthcare establishments produce airborne fungal spores and considerably increase the risk of exposure of immunosuppressed patients. It is necessary to reinforce protective measures, or even to implement specific precautions, during this critical phase. The aim of these precautions is to protect both those areas, which are susceptible to dust, and patients at risk of a fungal infection particularly invasive aspergillosis. When construction works are planned in healthcare establishments, the first step consists in the characterisation of the environmental fungal risk and the second one in proposing risk management methods. It is then essential to establish impact indicators in order to evaluate the risk management precautions applied. The working group promoted by the French societies of medical mycology and hospital hygiene (SFMM & SF2H) details here both environmental and epidemiological impact indicators that can be used.

Molecular characterization of glycopeptide-resistant enterococci from hospitals of the picardy region (france).

We studied 138 glycopeptide-resistant enterococci (GRE) strains, consisting of 131 glycopeptide-resistant Enterococcus faecium (GREfm) and 7 glycopeptide-resistant Enterococcus faecalis (GREfs). The GREfm strains were resistant to penicillin, ampicillin, vancomycin, and teicoplanin, while the GREfs strains were only resistant to vancomycin and teicoplanin. The van A gene was the only glycopeptide determinant present in all GRE isolates investigated. Genes coding for Hyl and Hyl+ Esp were detected in 39 (29.8%) and 92 (70.2%) of the 131 GREfm isolates, respectively. Three of the 7 GREfs were positive for gelE+asa 1 genes, 3 for gel E gene, and 1 for asa 1 gene. The genetic relationship between the 138 GRE was analyzed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). GREfm isolates were clustered in a single genogroup (pulsotype A), and GREfs were clustered in six genogroups (pulsotypes B-G). Among the isolates investigated by MLST, only 18 PCR products were sequenced (12 E. faecium and 6 E. faecalis), and 9 sequence types (STs) were identified.

[A sensitive, specific and predictive isolation medium developed for Stenotrophomonas maltophilia study in healthcare settings]. / La mise au point d'un milieu sensible, spécifique et prédictif de recherche de Stenotrophomonas maltophilia dans l'environnement des soins.

BACKGROUND: Stenotrophomonas maltophilia (Smalto) is a prominent nosocomial pathogen, commonly isolated in the hospital environment. Multiple Smalto nosocomial outbreaks have been linked to contaminated water sources. This study aimed to develop a medium able to ease healthcare environment Smalto isolation. METHODS: Financed, from March 2007 to June 2008, by a university hospital of Amiens' clinical research program, this study allowed Stenotrophomonas maltophilia selective medium with coloured indicator (SM2i) development. SM2i is constituted of Mueller Hinton agar (MH), maltose, DL-methionine, bromothymol blue. The mixture sterilized is refreshed at 50 degrees C, its pH adjusted to 7.1, and render selective by addition of vancomycin, imipenem and amphotericin B. Then, SM2i agar is sunk into 90 cm diameter Petri dish dated and stored at 4 degrees C for 4 weeks. SM2i is developed using Pasteur Institute culture type collection (CIP) strains of Smalto, Burkholderia cepacia, Pseudomonas aeruginosa (Psa) and a Smalto strain of our hygiene laboratory collection. It was validate on Psa imipenem-resistant and Enterococcus faecium vancomycin-resistant strains, then, tested on cold water first jet and faucet cotton-swabs samples. SM2i tests were made in comparison with the MH agar, MH agar plus four paper disks loaded 10 microg of imipenem and Cetrimed agar. Its sensitivity, specificity, positive (PPV) and negative (NPV) predictive values, accuracy, likehood-ratio (LR) and Youden index have been determined. RESULTS: SM2i agar is better in culturing Smalto test-strains. On SM2i, Smalto colonies are smooth, round, greeny, olive or lime green, have a green olive centre with a peripheral lighter or a dark green centre with an olive green suburb surrounded by a blue halo. SM2i is a selective, specific, predictive, accurate medium to search for Smalto in healthcare environment. In 122 pairs of cold water first jet and taps cotton-swabs samples, Smalto was isolated from 14.8% of water samples, 10.7% of cotton-swabs samples. It was isolated alone in 6.6% of water samples and 2.5% of swab samples. Thus, smalto has biocontaminated 17.2% of cold water taps. Compared to MH agar, SM2i sensitivity, specificity, PPV, NPV, accuracy, LR were 100, 100, 100, 100, 100% and infinity, and 87.5, 100, 100, 98.1, 98.4% and infinity for water and cotton-swabs samples respectively. CONCLUSION: SM2i is a selective, specific, predictive medium which can allow easily isolating and identifying accurately Smalto in environmental samples. Its evaluation on clinical samples is on going.

[Stenotrophomonas maltophilia and Pseudomonas aeruginosa water-associated microbiologic risk assessment in Amiens' University Hospital Centre]. / Evaluation des risques microbiologiques hydriques associés à Stenotrophomonas maltophilia et Pseudomonas aeruginosa au CHU d'Amiens.

BACKGROUND: Pseudomonas aeruginosa (Psa) and Stenotrophomonas maltophilia (Smalto) are major opportunistic waterborne pathogens causing hospital-acquired infections. This study aimed to assess the biocontamination level of cold water used in Amiens' university hospital wards, from March to June 2008. METHODS: We cultivated 122 pairs of cold water first jet and taps cotton-swabs on Cetrimide agar for Psa, on Stenotrophomonas maltophilia selective medium with coloured indicator (SM2i) for Smalto, on Mueller Hinton agar used as isolation medium reference for both, 48h at 30 degrees C. Data analysed with Epi-Info 6.04dFr were compared with chi(2) test, significant at p<.05. RESULTS: Psa and Smalto were isolated in 26.2 and 14.8% of water samples and in 21.3 and 10.7% of swab samples respectively. They were associated in 11.5% of water samples and 5% of swab samples. Psa was alone in 13.1% of water samples and 7.4% of swab samples whereas Smalto was found in 6.6% of water and 2.5% of swabs. Psa and Smalto were isolated from 14.8% of water samples and 8.2% of swab samples of the same tap. Finally, respectively 35.2 and 17.2% of the cold water taps were biocontaminated by Psa and Smalto. In fact, microbiologic water taps contamination risk was two-fold higher for Psa than for Smalto, p<.001, without variation between wards. CONCLUSION: Sm2i and Cetrimide are suited and efficient medium respectively for Smalto and Psa isolation. Cold-water samples are sufficient for waterborne pathogens biocontamination risk appraisal. Our results urged healthcare workers on efficient water fittings microbiologic risk control to prevent healthcare associated waterborne infections, notably due to Psa and Smalto.

Successive emergence of extended-spectrum beta-lactamase-producing and carbapenemase-producing Enterobacter aerogenes isolates in a university hospital.

Sixty-two clinical isolates of Enterobacter aerogenes resistant to expanded-spectrum cephalosporins were collected between July 2003 and May 2005. Among these isolates, 23 (37.1%) were imipenem (IPM) susceptible, and 39 (62.9%) were IPM insusceptible, of which 89.7% (35/39) were resistant and 10.3% (4/39) were intermediate. Isolate genotypes were compared by pulsed-field gel electrophoresis. Of 62 isolates, 48 belonged to epidemic pulsotype A (77.4%). This pulsotype included 37.5% and 58.4% of beta-lactam phenotypes b and a, respectively. Nine isolates (14.5%) belonged to pulsotype E, which included 22.3% and 77.7% of phenotypes b and a, respectively. The beta-lactamases with pIs of 5.4, 6.5, 8.2, and 8.2 corresponded to extended-spectrum beta-lactamases (ESBLs) TEM-20, TEM-24, SHV-5, and SHV-12, respectively. Of 39 IPM-insusceptible E. aerogenes isolates, 26 (66.6%) were determined to be metallo-beta-lactamase producers, by using a phenotypic method. Of these isolates, 24 harbored a bla(IMP-1) gene encoding a protein with a pI of >9.5, and two carried the bla(VIM-2) gene encoding a protein with a pI of 5.3, corresponding to beta-lactamases IMP-1 and VIM-2, respectively. The remaining 13 (33.4%) isolates were negative for the bla(IMP-1) and bla(VIM-2) genes but showed an alteration of their outer membrane proteins (OMPs). Ten of these isolates produced the two possible OMPs (32 and 42 kDa), with IPM MICs between 8 and 32 microg/ml, and three others produced only a 32-kDa OMP with IPM MICs >32 microg/ml. This work demonstrates that, in addition to resistance to expanded-spectrum cephalosporins, IPM resistance can occur in ESBL-producing E. aerogenes isolates by carbapenemase production or by the loss of porin in the outer membrane.

[Extended-spectrum betalactamases producing Escherichia coli: a new health-care associated infection threat?]. / Escherichia coli producteurs de bêtalactamases à spectre étendu: de nouvelles menaces nosocomiales ?

OBJECTIVES: Study the health-care associated infection risk due to Extended-Spectrum Betalactamases Producing Escherichia coli (ESBL Esc) isolated from diagnostic samples. METHODS: Descriptive, longitudinal and prospective study of 104 diagnostic isolates of ESBL Esc, one per patient, identified in Amiens university hospital between February 1999 and December 2005. Patients (sex, age, contamination risk factor, antecedent hospitalization) and microbiological data were progressively collected, entered into EPI INFO 6.04dFr software (ENSP, France) database, and compared using the chi-square test and Wilcoxon rank sum test, as appropriate. A p value of less than 0.05 was considered significant. RESULTS: Diagnostic ESBL Esc isolates raised, per 1000 isolates of Esc, from 1.2 in 1999 to 6 in 2005. Global and acquired isolates number of ESBL Esc varied from 7 and 3 in 2002 to 25 and 19 in 2003 (P=0.22). ESBL Esc global and acquired incidence per 10(5) patient-days were, 0.8 and 0.6 in 1999 and 4.99 and 3.4 in 2005 (P<10(-6)), but rose from 0.6 acquired isolate in 2002 to 3.9 in 2003 (P=0.002). ESBL Esc, isolated from urines, stools, pulmonary, blood and surgical site samples of patients of>/=65 years aged (68.3%), were imipenem and latamoxef sensitive. Their acquisition risk factors found were hospitalization during the last 6 month period (40/104) and transfer from other institutions (20/104). CONCLUSION: ESBL Esc isolates, among ESBL-producing Enterobacteriaceae, constitute an escalating health-care associated risk in our institution. The research at admission time of ESBL-producing Enterobacteriaceae, mainly in acute geriatric wards, strict isolation precaution and hand hygiene observance, rational antibiotic usage, are the key actions to control their cross transmission. Nonetheless, other studies are needed to determine whether we are in front of an ESBL Esc new clone emergence.

[Case control study of extended-spectrum betalactamase producing Serratia marcescens outbreak in a paediatric intensive care unit]. / Etude cas-témoins d'une épidémie à Serratia marcescens dans un service de réanimation pédiatrique.

OBJECTIVES: To identify patient-related risk factors of infection and ways of transmission of extended-spectrum betalactamase (ESBL) producing Serratia marcescens in the paediatric intensive care unit (PICU) of Amiens university hospital (France) between June and July 2002. METHODS: Five cases (four pulmonary infected and one stool contaminated symptom-free neonates) and 35 controls, admitted in the PICU, are included. S. marcescens ESBL analysed are isolated from respiratory tract and faecal samples for cases and urine and pus samples from two non-paediatric other patients. Univariate and multivariate analysis are performed on EPI INFO 6.04 dFr and SPSS 11.0.1. RESULTS: S. marcescens ESBL infections or colonisations rate is 12.5% [4.7-27.6]. The incidence is 8.8 [6.7-11.6] per 1000 hospital-stay days. By univariate analysis, cases and controls don't differ with respect of age, sex, and weight at admission or preterm delivery. Cases don't have more often invasive nursing care than controls. But, they were intubated (P <0.03) and hospitalised (P <0.03) for a longer time than controls. Linear regression analysis showed that duration of intubation was independent predictor of acquisition of S. marcescens ESBL (P <0.008). S. marcescens ESBL strains implicated in pulmonary infections, showed the same pattern of multidrug resistant and ERIC-PCR profile. This clone differs from others isolated from stool or other samples from other hospital wards. CONCLUSION: As S. marcescens cross-colonization appears to be due to lake of hand hygiene and asepsis during invasive nursing care, reinforcing hygiene measures permit to contain the outbreak.

Prevalence of Paramphistomum daubneyi infection in cattle in central France.

Investigations at the slaughterhouse of Limoges (Central France) were performed in 1994-1996 to determine the prevalence of Paramphistomum daubneyi infection in cattle. In 1994/1995, higher prevalences of P. daubneyi were recorded in May, October, and January. In 1996, smaller variations were recorded, and prevalence in April and May was higher. The prevalence of P. daubneyi in summer was lower. Breed and age of cattle were not significantly associated to P. daubneyi infection, but females were significantly more infected than males. P daubneyi infection was related to that of Fasciola hepatica, but not to that of Dicrocoelium lanceolatum.

[Pneumococcus: an unusual agent of maternal-fetal infection]. / Le pneumocoque: un agent inhabituel d'infection materno-foetale.

BACKGROUND: Since the advent of antibiotics, Streptococcus pneumoniae has become a very unusual agent of materno-fetal infection. We report two cases. CASE REPORTS: In case n(o) 1, early neonatal meningitis was caused by a penicillin-resistant strain. In case n(o) 2, the mother developed meningitis 16 days after delivery. In both cases, premature rupture of the membranes in otherwise asymptomatic mothers was the initial event and outcome was favorable under amoxicillin alone. The pneumococci were easily recovered fom placental, amniotic and neonatal samples, less easily from the maternal samples. DISCUSSION: The increasing prevalence of penicillin-resistant pneumococci is an emerging problem of real concern. It might increase the very low present incidence of pneumococcal neonatal materno-fetal infection which is a particularly serious infection with up to 60% neonatal mortality. Any type of S. pneumoniae infection, or even colonization, occurring in the peripartum should prompt adequate treatment and suggests considering first line vancomycin for the newborn.

Seroprevalence of cysticercosis in Bénin.

We report the results of a seroepidemiological study on the prevalence of cysticercosis in Bénin. Cluster sampling at 3 levels was performed in the 6 départements (Atacora, Borgou, Zou, Mono, Atlantique and Oueme) and 2625 serum samples, from 1329 adult females and 1296 adult males, were collected. Antibodies against Taenia solium cysticerci were first searched for by enzyme-linked immunosorbent assay and the 41 seropositive samples were then examined by enzyme-linked electroimmunotransfer blot assay (EITB). Thirty-five samples gave positive results in the EITB. The overall seroprevalence of cysticercosis was therefore 1.3% (95% confidence interval [95% CI] 0.9-1.9). The seroprevalence was 1.9% in males (95% CI 1.2-2.7) and 0.8% (95% CI 0.4-1.5) in females (P < 0.05). A progressive increase in seroprevalence with increasing age was found. The highest seroprevalences were observed in Atacora and Atlantique, 2 non-Muslim départements (3.3% and 3.0%, respectively). This study demonstrated the public health importance of cysticercosis in Bénin.

[Seroprevalence of HBV, HCV and HDV hepatitis markers in 500 patients infected with the human immunodeficiency virus]. / Séroprévalence des marqueurs des virus des hépatites B, C et D chez 500 patients infectés par le virus de l'immunodéficience humaine.

The serological status for both hepatitis B, C and D viruses was analyzed for 500 HIV seropositive patients and for 1037 of a control group. The prevalence was 31.4% for anti HCV, 13.8% for HBs Ag and 69.0% for one or more HBV markers in HIV positive patients and respectively 2.5%, 2.7% and 13.1% in control group. The markers for hepatitis D were founded among 21% of the HBs Ag carriers (patients and control group), correlated with drug i.v. use. The prevalence of anti-HCV was 71.6% in subjects who had blood-borne HIV infection and 1.5% in those with sexually acquired infection. The prevalence in control group was 10.2% and 1.7% respectively according to the same risk factors. The prevalence of HBs Ag was higher among HIV positive patients with sexual risk (17.5%) than with blood exposition (9.9%) and a variation in the same direction is observed in control group (3% v.s. 1%). The relation between markers for hepatitis B and hepatitis C was negative.

[Seroprevalence of cysticercosis in the lacustrian community of Vekky, Atlantic district (Benin)]. / Séroprévalence de la cysticercose dans la commune lacustre de Vekky, département de l'Atlantique (Bénin).

Cysticercosis is a parasitic disease due to the infection of man with Cysticercus cellulose, the larva of Taenia solium. This disease is frequent in countries with low socio-economic development and is linked to sanitary conditions. The aim is to assess the seroprevalence of cysticercosis in the lacustrine vicinity of Vekky, located on the lake Nokoué, District Atlantic, south Benin, an epidemiological survey was undertaken in April and May 1994. The lacustrian vicinity of Vekky comprises 12 villages including 16, 142 inhabitants. Population has been sampled using cluster sampling method (n = 30) at two levels (village and household). The whole samples consisted in 319 adults (123 females and 196 males, mean of ages: 32.8 +/- 18.3). Titration of cysticercosis antibodies has been made using ELISA. Eleven patients (3 females and 8 males) showed a positive ELISA response for cysticercosis, i.e. the seroprevalence of cysticercosis was 3.5% (CI 95%:1.3-8%). There was no significant difference according to age and sex. The seroprevalence of cysticercosis reached 9.1% in patients who presented history of epilepsy. We failed to find any linkage between seropositivity and i) clinical history of epilepsy or taeniasis, or ii) several studied environmental factors such as consumption of pork, wandering of pigs, lack of veterinary supervision, religion and occupation. Human seroprevalence of cysticercosis reaches 3.5% in vicinity of Vekky, which denotes a high endemic level. Further epidemiological studies are necessary to precise the factors involving cysticercosis in this area.