Impact of a new hospital with close to 100% single-occupancy rooms on environmental contamination and incidence of vancomycin-resistant Enterococcus faecium colonization or infection: a genomic surveillance study.

BACKGROUND: Vancomycin-resistant Enterococcus faecium (VRE) is a leading cause of nosocomial infection, driven by its ability to spread between patients and persist in the hospital environment. AIM: To investigate the impact of a long-established cardiothoracic hospital moving to new premises with close to 100% single-occupancy rooms on the rates of environmental contamination and infection or colonization by VRE. METHODS: Prospective environmental surveillance for VRE was conducted at five time-points between April and November 2019, once in the original building, and four times in the new building. Incidence rate ratios (IRRs) of VRE infection/colonization were determined for the one-year period before and after the hospital move, and compared to a nearby hospital. FINDINGS: In the original location, the first environmental screen found 29% VRE positivity. The following four screens in the new location showed a significant reduction in positivity (1-6%; P<0.0001). The VRE infection/colonization rates were halved in the new location (IRR: 0.56; 95% confidence interval: 0.38-0.84), compared to the original location, contrasting with an increase in a nearby hospital (1.62; 1.17-2.27) over the same time-period. Genomic analysis of the environmental isolates was consistent with reduced transmission in the new hospital. CONCLUSION: The use of single-occupancy rooms was associated with reduced environmental contamination with VRE, and lower transmission and isolation of VRE from clinical samples. The cost-effectiveness of single-occupancy room hospitals in reducing healthcare-associated infections should be reassessed in the context of operational costs of emerging pandemic and increasing antimicrobial resistance threats.

Serotype 3 is a common serotype causing invasive pneumococcal disease in children less than 5 years old, as identified by real-time PCR.

Serotype 3 is one of the most often detected pneumococcal serotypes in adults and it is associated with serious disease. In contrast, the isolation of serotype 3 by bacterial culture is unusual in children with invasive pneumococcal disease (IPD). The purpose of this study was to learn the serotype distribution of IPD, including culture-negative episodes, by using molecular methods in normal sterile samples. We studied all children<5 years of age with IPD admitted to two paediatric hospitals in Catalonia, Spain, from 2007 to 2009. A sequential real-time polymerase chain reaction (PCR) approach was added to routine methods for the detection and serotyping of pneumococcal infection. Among 257 episodes (219 pneumonia, 27 meningitis, six bacteraemia and five others), 33.5% were identified by culture and the rest, 66.5%, were detected exclusively by real-time PCR. The most common serotypes detected by culture were serotypes 1 (26.7%) and 19A (25.6%), and by real-time PCR, serotypes 1 (19.8%) and 3 (18.1%). Theoretical coverage rates by the PCV7, PCV10 and PCV13 vaccines were 10.5, 52.3 and 87.2%, respectively, for those episodes identified by culture, compared to 5.3, 31.6 and 60.2% for those identified only by real-time PCR. Multiplex real-time PCR has been shown to be useful for surveillance studies of IPD. Serotype 3 is underdiagnosed by culture and is important in paediatric IPD.

Characterization of avocado honey (Persea americana Mill.) produced in Southern Spain.

Plant origin, physicochemical parameters and composition were analysed to characterize the avocado honey (Persea americana Mill.) from Andalusia (Southern, Spain). Ashes content, total polyphenol, and electrical conductivity corresponded to these of a typical dark honey (>80 mm scale Pfund). Regarding mineral elements, K was predominant, followed by P and Mg. Antioxidant and invertase activities presented some desirable values. In the 20 analysed samples, 48 pollen types corresponding to 33 families were identified. Avocado pollen was found in high variability (13-58%). At least a 20% was suggested to guarantee the authentic avocado honey. Perseitol, sugar-alcohol identified only in avocado honey, fundamentally contributes to distinguish this kind of honey. The content varied between 0.31 and 1.56 g/100 g. The correlation between perseitol and avocado pollen was found to be significant. A minimum concentration of 0.30 g/100 g of perseitol is suggested to characterize the proposed monofloral avocado honey.

Population-based analysis of ocular Chlamydia trachomatis in trachoma-endemic West African communities identifies genomic markers of disease severity.

BACKGROUND: Chlamydia trachomatis (Ct) is the most common infectious cause of blindness and bacterial sexually transmitted infection worldwide. Ct strain-specific differences in clinical trachoma suggest that genetic polymorphisms in Ct may contribute to the observed variability in severity of clinical disease. METHODS: Using Ct whole genome sequences obtained directly from conjunctival swabs, we studied Ct genomic diversity and associations between Ct genetic polymorphisms with ocular localization and disease severity in a treatment-naïve trachoma-endemic population in Guinea-Bissau, West Africa. RESULTS: All Ct sequences fall within the T2 ocular clade phylogenetically. This is consistent with the presence of the characteristic deletion in trpA resulting in a truncated non-functional protein and the ocular tyrosine repeat regions present in tarP associated with ocular tissue localization. We have identified 21 Ct non-synonymous single nucleotide polymorphisms (SNPs) associated with ocular localization, including SNPs within pmpD (odds ratio, OR = 4.07, p* = 0.001) and tarP (OR = 0.34, p* = 0.009). Eight synonymous SNPs associated with disease severity were found in yjfH (rlmB) (OR = 0.13, p* = 0.037), CTA0273 (OR = 0.12, p* = 0.027), trmD (OR = 0.12, p* = 0.032), CTA0744 (OR = 0.12, p* = 0.041), glgA (OR = 0.10, p* = 0.026), alaS (OR = 0.10, p* = 0.032), pmpE (OR = 0.08, p* = 0.001) and the intergenic region CTA0744-CTA0745 (OR = 0.13, p* = 0.043). CONCLUSIONS: This study demonstrates the extent of genomic diversity within a naturally circulating population of ocular Ct and is the first to describe novel genomic associations with disease severity. These findings direct investigation of host-pathogen interactions that may be important in ocular Ct pathogenesis and disease transmission.

Large-scale whole genome sequencing of M. tuberculosis provides insights into transmission in a high prevalence area.

To improve understanding of the factors influencing tuberculosis transmission and the role of pathogen variation, we sequenced all available specimens from patients diagnosed over 15 years in a whole district in Malawi. Mycobacterium tuberculosis lineages were assigned and transmission networks constructed, allowing ≤10 single nucleotide polymorphisms (SNPs) difference. We defined disease as due to recent infection if the network-determined source was within 5 years, and assessed transmissibility from forward transmissions resulting in disease. High-quality sequences were available for 1687 disease episodes (72% of all culture-positive episodes): 66% of patients linked to at least one other patient. The between-patient mutation rate was 0.26 SNPs/year (95% CI 0.21-0.31). We showed striking differences by lineage in the proportion of disease due to recent transmission and in transmissibility (highest for lineage-2 and lowest for lineage-1) that were not confounded by immigration, HIV status or drug resistance. Transmissions resulting in disease decreased markedly over time.

Factors affecting the formation of alkylpyrazines during roasting treatment in natural and alkalinized cocoa powder.

The cocoa roasting process at different temperatures (at 125 and 135 degrees C for 3 min, plus 44 and 52 min, respectively, heating-up times) was evaluated by measuring the initial and final free amino acids distribution, flavor index, formol number, browning measurement, and alkylpyrazines content in 15 cocoa bean samples of different origins. These samples were also analyzed in manufactured cocoa powder. The effect of alkalinization of cocoa was studied. Results indicated that the final concentration and ratio of tetramethylpyrazine/trimethylpyrazine (TMP/TrMP) increased rapidly at higher roasting temperatures. The samples roasted with alkalies (pH between 7.20 and 7.92), such as sodium carbonate, or potassium plus air injected in the roaster during thermal treatment, exhibited a greater degree of brown color formation, but the amount of alkylpyrazines generated was adversely affected. The analysis of alpha-free amino acids at the end of the roasting process demonstrated the importance of the thermal treatment conditions and the pH values on nibs (cocoa bean cotyledons), liquor, or cocoa. Higher pH values led to a lower concentration of aroma and a higher presence of brown compounds.

Liquid chromatographic determination of tocopherols and tocotrienols in vegetable oils, formulated preparations, and biscuits.

A precise and selective liquid chromatographic procedure for determining tocopherol and tocotrienol isomers in vegetable oils, formulated preparations, and biscuits was developed and validated. The proposed method quantitates vitamin E in better conditions of recoverability and reproducibility than the standard saponification procedure. Tocopherols and tocotrienols were extracted in hexane from vegetable oils, passed through a silica Sep-pak, chromatographed on a mu-Bondapak C18 column with a mobile phase of methanol-water (95 + 5, v/v), identified at 292 nm, and detected with fluorescence procedure (excitation 296 nm, and emission 330 nm). The correlation coefficient on the calibration curve was 0.9995 over the range of 0.1 to 100 microg/mL. Overall recovery of vitamin E isomers was 93%; coefficients of variation for intra- and interday precision, < 2.25%. The results obtained from extraction methods 1 (with saponification) and 2 (without saponification) were compared by ANOVA test. Significant differences appeared between vitamin E isomers (p < or = 0.05).

[Bupropion combined with nicotine patches in smoking cessation]. / Asociación de bupropión y parches de nicotina como terapia para dejar de fumar.

Smoking cessation is a first-line treatment for patients with bronchial and pulmonary diseases. Various strategies have been developed to help patients quit. Bupropion, a drug initially developed as an antidepressant, has recently been shown to have effects that increase the ability of a smoker to quit. This descriptive study, enrolling 86 patients who volunteered for a smoking cessation program, assesses the use of 300 mg of bupropion over a 4-week period combined with 12 weeks of nicotine patch application at doses that were lowered every 4 weeks. Abstinence was achieved by 69% of patients after 6 months of follow-up and no significant side effects were described. The percentage decreased to 58.6% after one year of follow-up. No significant differences were found between success in quitting in this study and either the number of prior attempts to quit or concomitant respiratory disease. We conclude that bupropion combined with transdermal nicotine is a good option to aid patients to achieve smoking cessation.

[Smoking cessation and bupropion: anxiety and depression as predictors of therapeutic efficacy]. / Deshabituación tabáquica y bupropión: la ansiedad y la depresión como índices de eficacia terapéutica.

UNLABELLED: Smoking and depression are related. Bupropion, the first non-nicotinic drug that is an effective treatment in smoking cessation, is a tricyclic antidepressant that inhibits neuronal uptake of serotonin, dopamine and norepinephrine in the thalamic nuclei. OBJECTIVE: To assess if certain personality factors (anxiety or depression) might predict the efficacy of bupropion for smoking cessation. METHOD: The study was carried out in two smoking cessation clinics in Madrid and Barcelona. Fifty patients (21 men) declaring the desire to quit smoking were enrolled. Their mean age was 43.6 years (SD 8.75). The patients were treated with 300 mg of bupropion per day for one month and expired CO was monitored for 6 months. Personality factors were assessed on a hospital anxiety and depression scale (HADS). We evaluated whether there was a significant difference in HADS scores for patients who were still not smoking after 6 months and those who had not managed to quit. RESULTS: The 50 patients were smokers of a mean 39 packs per year (SD 17.82) and had mean scores of 7.4 (SD 4.15) for anxiety and 5.8 (SD 3.93) for depression. Four patients (8%) were unable to complete the study. After one month, 28% of the patients smoked, after 3 months 56% smoked and after 6 months 58% still smoked. The patients who smoked during the first month had higher depression scores than did the non-smokers (p = 0.03). After 3 and 6 months the patients who had managed to continue not smoking were those who had higher anxiety scores than did those who still smoked (p = 0.0052 at 3 months and p = 0.017 at 6 months). CONCLUSION: Patients who responded better to treatment with bupropion after 6 months of follow-up were those with higher anxiety scores on the HADS. Depression levels influenced outcome only during the first month.

Study on propolis quality from China and Uruguay.

The composition, bacteriostatic and ROO*-scavenging potential activities of fifteen propolis samples from various botanic and geographic origins were determined to obtain objective information related to propolis quality. Variance analysis showed significant differences (p < or = 0.05) in the contents of polyphenols, flavonoids and active components between fresh and aged propolis. The state of the product (fresh or aged) could be differentiate by using flavonoid pattern and biological activities. A minimum propolis concentration of 80 microg/ml was required inhibit Bacillus subtilis and Staphylococcus aureus while 800 microg/ml was required to inhibit Escherichia coli using fresh propolis. Aged propolis inhibit B. subtilis and S. aureus at concentration of 100 microg/ml and E. coli at 1000 microg/ml. A minimum flavonoids percentage of 18 g/100 g and a maximum ROO*-scavenging potential activity of 4.3 microg/ml were determined in fresh propolis. Flavonoids levels in aged propolis were approximately 20% lower than in fresh propolis. A maximum flavonoids percentage of 19.8 g/100 g and a ROO*-scavenging potential activity between 5.7 to 6.4 microg/ml in aged propolis were quantified. Another objective was to assess the use of ROO*-scavenging potential activity in propolis quality.

Role of the subunits of the energy-transducing adenosine triphosphatase from Micrococcus lysodeikticus membranes studied by proteolytic digestion and immunological approaches.

An energy-transducing adenosine triphosphatase (ATPase, EC 3.6.1.3) that contains an extra polypeptide (delta) as well as three intrinsic subunits (alpha, beta, gamma) was purified from Micrococcus lysodeikticus membranes. The apparent subunit stoichiometry of this soluble ATPase complex is alpha 3 beta 3 gamma delta. The functional role of the subunits was studied by correlating subunit sensitivity to trypsin and effect of antibodies raised against holo-ATPase and its alpha, beta and gamma subunits with changes in ATPase activity and ATPase rebinding to membranes. A form of the ATPase with the subunit proportions 1.67(alpha):3.00(beta:0.17(gamma) was isolated after trypsin treatment of purified ATPase. This form has more than twice the specific activity of native enzyme. Other forms with less relative proportion of alpha subunits and absence of gamma subunit are not active. Of the antisera to subunits, only anti-(beta-subunit) serum shows a slight inhibitory effect on ATPase activity, but its combination with either anti-(alpha-subunit) or anti-(gamma-subunit) serum increases the effect. The results suggest that beta subunit is required for full ATPase activity, although a minor proportion of alpha and perhaps gamma subunit(s) is also required, probably to impart an active conformation to the protein. The additional polypeptide not hitherto described in Micrococcus lysodeikticus ATPase had a molecular weight of 20 000 and was found to be involved in ATPase binding to membranes. This 20 000-dalton component can be equated with the delta subunit of other energy-transducing ATPases and its association with the (alpha, beta, gamma) M. lysodeikticus ATPase complex appears to be dependent on bivalent cations. The present results do not preclude the possibility that the gamma subunit also plays a role in ATPase binding, in which, however, the major subunits do not seem to play a role.

Evaluation of Smoky Taste in Cocoa Powder.

The aim of this study was to determine the levels of nine phenols (phenol, guaiacol derivatives, xylenols, and cresols) by using a GC/MS technique in smoked samples under control, commercial samples taken from contaminated batches, and uncontaminated samples. The smoky taste of cocoa powder may have two origins: drying and storage. The sample was steam distilled, extracted with ethyl ether, concentrated, and chromatographed on a 50-m OV-351 fused silica column. The physicochemical data obtained were related to sensory evaluation and submitted to multivariate distribution (Mahalanobis distance) and Pearson Chi-squared test. The discriminatory phenols identified were phenol, 3-methylphenol (m-cresol), 2,3-dimethylphenol (2,3-xylenol), 3-ethylphenol, 4-ethylphenol, and total phenols. The results obtained imply that uncontaminated smoked cocoa samples should have the following maximum concentrations: phenol, 2 mg/kg; 3-methylphenol, 0.9 mg/kg; 2,3-dimethylphenol, 0.55 mg/kg; 3-ethylphenol, 0.90 mg/kg; 4-ethylphenol, 0.70 mg/kg; and total phenols, 9.6 mg/kg.

Enumeration of Hydrogen Sulfide-Producing Bacteria from Anaerobically Packaged Pork.

The influence of medium composition, pH, incubation time and gaseous atmosphere during incubation on enumeration of H2S-producing bacteria from anaerobically packaged pork was determined. Samples of anaerobically packaged pork were plated and H2S-producing bacteria isolated. Stock cultures of these isolates were prepared, diluted and pour-plated with Lead Acetate Agar (LAA) containing various concentrations of cysteine (0.001 to 0.005%) at different pH values (5.0 to 8.0). The inoculated plates were incubated at 21°C for various periods (3 to 6 d) under different gaseous atmospheres (N2, CO2 or mixtures of both). The conditions resulting in optimized recovery and enumeration of H2S-producing bacteria from anaerobically packaged pork consisted of pour-plating the isolates with LAA to which 0.003 to 0.005% cysteine was added and pH adjusted to 7.5-8.0 plus incubation in an atmosphere of 5%CO2-95%N2 for 5 to 6 d at 21°C.