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.

Design, synthesis, and characterization of novel aminoalcohol quinolines with strong in vitro antimalarial activity.

Malaria is the fifth most lethal parasitic infections in the world. Herein, five new series of aminoalcohol quinolines including fifty-two compounds were designed, synthesized and evaluated in vitro against Pf3D7 and PfW2 strains. Among them, fourteen displayed IC50 values below or near of 50.0 nM whatever the strain with selectivity index often superior to 100.17b was found as a promising antimalarial candidate with IC50 values of 14.9 nM and 11.0 nM against respectively Pf3D7 and PfW2 and a selectivity index higher than 770 whatever the cell line is. Further experiments were achieved to confirm the safety and to establish the preliminary ADMET profile of compound 17b before the in vivo study performed on a mouse model of P. berghei ANKA infection. The overall data of this study allowed to establish new structure-activity relationships and the development of novel agents with improved pharmacokinetic properties.

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.

ß-Alkylated oligomaltosides as new alternative preservatives: antimicrobial activity, cytotoxicity and preliminary investigation of their mechanism of action.

AIM: The need for developing efficient and safe alternatives to parabens has been growing in the cosmetic and pharmaceutical industries. To this end, the antimicrobial activities and safety of methyl-ß-d-maltoside, methyl-ß-d-maltotrioside and their dodecyl homologues were investigated. METHODS AND RESULTS: Antimicrobial activities of methyl- and dodecyl-ß-d-oligomaltoside have been tested on European pharmacopoeia reference strains. When effective, minimal inhibitory concentrations ranged from 32 to 1024 mg l(-1) . Methyl derivatives exhibited greater antibacterial properties, while their dodecyl homologues were more active on fungal strains. To elucidate the mechanism of action of methyl compounds, enzymatic inhibition assays of key maltose metabolism enzymes have been conducted. Methyl-ß-maltotrioside (MeG3) was found to be effective in inhibiting microbial glucoamylase and α-amylase. MeG3 and dodecyl-ß-maltotrioside cytotoxicity were also checked on HepG2 cells and were found to be low, compared with benzalkonium chloride or parabens. CONCLUSION: Methyl- and dodecyl-ß-maltoside or maltotrioside were found to be active against reference strains used for preservatives efficacy testing. Methyl derivatives could act through an interesting inhibition of the microbial enzymatic metabolism. SIGNIFICANCE AND IMPACT OF THE STUDY: Given their very simple chemical structure, low toxicity and good antimicrobial activity, methyl-ß-d-oligomaltosides could represent a valuable alternative to currently used preservatives such as parabens.

[Establishment of the intestinal microflora and regulation of bacterial translocation after caffeine citrate treatment during postnatal period in rat]. / Implantation de la microflore intestinale et régulation de la translocation bactérienne sous l'effet d'un traitement au citrate de caféine chez le raton en période néonatale.

To relieve respiratory problems such as apnea in newborns, caffeine citrate is the drug of choice because of its good tolerance and therapeutic index. However, its impact on the intestinal microbial ecosystem and on bacterial translocation in the neonatal period remains insufficiently investigated. Therefore, the objective of this study was to evaluate the effects of caffeine citrate on the establishment of the intestinal microflora and bacterial translocation in rats from birth to the 30th day of life. Newborn Wistar rats were divided into four groups of 14 animals, each subdivided into a control group receiving a placebo (12mL tap water/kg/day) and another treated with caffeine citrate (12mg/kg/day). The animals were nursed by their mothers and weighed daily. A group of 14 rats was killed at birth and after 10, 20, or 30 days of life. Organs in which translocation was assessed (liver, lungs, spleen, and kidneys) and various fragments of intestine (duodenum, jejunum, ileum, and colon) were surgically removed. The bacteriological analysis performed involved enumeration of the total microflora, staphylococci, enterobacteria, and lactobacilli. From the 10th day, caffeine was shown to significantly decrease the weight of treated animals as compared with controls (P<0.05). However, caffeine treatment did not drastically alter the kinetics of establishment of the intestinal microflora as only enterobacteria were found to be significantly lower in any intestinal segment of the treated group (P<0.05). Moreover, from the 20th day of life, caffeine citrate significantly downregulated bacterial translocation of both Gram-positive and -negative bacteria (P<0.05). This preliminary study on the effects of treatment with caffeine citrate may open opportunities in clinical pediatrics; the treatment will remain partially effective in preventing bacterial translocation in the neonatal period.

In vitro antimalarial activity of ICL670: a further proof of the correlation between inhibition of ß-hematin formation and of peroxidative degradation of hemin.

Iron chelators such as deferiprone, deferoxamine (DFO) and ICL670 (deferasirox) have previously been shown to display in vitro and/or in vivo antimalarial activities. To gain further insight in their antimalarial mechanism of action, their activities on inhibition of ß-hematin formation and on both peroxidative and glutathione (GSH)-mediated degradation of hemin were investigated. Neither deferiprone nor DFO were able to inhibit ß-hematin formation while ICL670 activity nearly matched that of chloroquine (CQ). Peroxidative degradation of hemin was also only strongly inhibited by both CQ and ICL670, the latter being significantly more efficient at pH 5.2. All iron chelators displayed minor, if any, inhibitory activity on GSH-mediated degradation of hemin. Discrepancies in the results obtained for the three iron chelators show that iron chelation is not the main driving force behind interference with heme degradation. Deferiprone, DFO and ICL670 share little structural community but both ICL670 and antimalarial ursolic acid derivatives (previously shown to block ß-hematin formation and the peroxidative degradation of hemin) have hydrophobic groups and hydroxyphenyl moieties. These similarities in structures and activities further back up a possible two-step mechanism of action previously proposed for ursolic acid derivatives (Mullié et al., 2010) implying (1) stacking of an hydrophobic structure to hemin and (2) additive protection of hemin ferric iron from H(2)O(2) by hydroxyphenyl groups through steric hindrance and/or trapping of oxygen reactive species in the direct neighborhood of ferric iron. These peculiar antimalarial mechanisms of action for ICL670 warrant further investigations and development.

Impact of colostomy on intestinal microflora and bacterial translocation in young rats fed with heat-killed Lactobacillus acidophilus strain LB.

A rat animal model of left colostomy was found to significantly impair the growth curve of rats. Assessment of the intestinal flora showed that colostomy mostly affects the cecal but not colonic microflora. Generally, the number of enterococci was increased in both ileum and cecum; cecal lactobacilli also rose, accounting for a promotion of lactic acid bacteria in colostomised rats. No significant differences between colostomised, laparotomised and control rats could be observed for the translocation of intestinal bacteria to internal organs of rats (i.e. spleen, kidneys, lungs or liver), whatever their diet. Heat-killed Lactobacillus acidophilus strain LB administration (dead probiotic bacteria) tended to exhibit a stimulatory effect on bifidobacteria, probably affecting the culture-medium fermentation substances included in the pharmaceutical product. This effect was abolished by laparotomy and colostomy. A trend towards a probiotic-like effect, not susceptible to colostomy, was also witnessed as counts of lactobacilli tended to increase in both cecum and colon of all animals fed with L. acidophilus LB.

Establishment and follow-up of bifidobacterial species in the gut of healthy bottle-fed infants of 1-4 months age.

Twenty-one healthy bottle-fed infants were screened monthly (1-4 months) for bifidobacteria in their stools. Bifidobacteria were detected by culture and isolates specified by PCR. Alternatively, direct PCR in undiluted fecal suspensions was carried out for detection of bifidobacteria under the cultural detection level. All infants harbored cultivable bifidobacteria throughout the study period. Beerens medium was shown to permit a better recovery of bifidobacteria than MRS and horse blood Columbia agar. Direct PCR detection proved valuable in detecting species for which no cultural isolate could be recovered since the species were under the cultural detection level. B. bifidum, B. longum-infantis and B. breve were confirmed as dominant and stable species in infant stools while B. adolescentis and B. catenulatum group exhibited unstable colonization profiles. A trend towards B. breve decrease began at month 3 while carriage of the B. catenulatum group and B. adolescentis was rising. This observation warrants further analysis to assess a possible switch occurring at month 3 in bottle-fed infants, between so-called infant and adult bifidobacterial species.

[Intestinal bacterial colonisation and translocation in C3H/HeJ conventional mice fed a unique dose of live or dead Bifidobacterium breve C50]. / Colonisation intestinale et translocation bactérienne chez la souris conventionnelle C3H/HeJ apres ingestion d'une dose unique de Bifidobacterium breve C50 vivantes ou mortes.

The efficiency and safety of use of Bifidobacterium breve C50 (BbC50), a potential probiotic, was assessed as regards intestinal microbial colonisation and bacterial translocation. A suspension of BbC50, containing 1-5 to 107-108 live bacteria, was fed to C3H/HeJ mice. The passage of live BbC50 was not demonstrated by culture either in the intestine or extra-intestinal organs. However, mice receiving the highest dose of live bacteria harbored more lactobacilli and less Bacteroides fragilis group in the cecum and colon when compared to control mice. Translocation of lactobacilli observed in the control group was not regulated by Bb50 feeding. Indeed, the spleen was significantly more frequently contaminated in mice fed BbC50, whatever the dose of live bacteria. The kidneys were also significantly more contaminated with lactobacilli in mice fed the highest dose of live Bb50. Moreover, higher dose of live BbC50 was associated with greater number of extra-intestinal contaminated organs. To conclude, BbC50 feeding induced a favorable balance in the mouse intestinal flora and was never found translocating, demonstrating its efficiency and safety of use. However, BbC50 seemed to interfere with the ability of lymphoid organs (e.g. the spleen) to eliminate translocating lactobacilli.

Partial characterization of bifidobacterium breve C50 cell-free whey compounds inducing modifications to the intestinal microflora.

Cell-free whey obtained from milk fermented with Bifidobacterium breve C50 (Bb C50) has been shown to modify the intestinal flora in humans and mice. Previous work showed that no antibiotic-like or barrier effect due to overgrowing bifidobacteria was implied in the microbial modifications. The present study was focused on characterizing the compounds and mechanisms involved. Protein, sugar, and enzymatic profiles of Bb C50 whey were therefore determined and compared to those of a whey unable to modify the intestinal flora of humans and mice. No remarkable difference was noted except for a higher lactosidase activity in Bb C50 whey. Various physical treatments were then applied to fractions of Bb C50 whey. Activity was assessed in C3H mice by analyzing changes in the intestinal flora balance throughout a 15-d administration of each treated whey. Heating at 80 degrees C and aerobic storage for 2 wk completely abolished Bb C50 whey activities. In contrast, the addition of a reducing agent (cysteine hydrochloride), either at the beginning of a 15-d aerobic storage or prior to administration, as well as preserved these activities. Susceptibility to heating and oxidation suggested that an enzyme might play a role in the induced microbial changes. Since the Bb C50 lactosidase was partly inactivated by the oxidative treatment, it could support the in vivo activity. The enzyme might reach the intestinal lumen and partly degrade substrates, such as mucins, usually used by the intestinal flora. The released molecules might then favor the development of a new microbial balance.