Verification and application of qPCR and viability-qPCR for Legionella monitoring in evaporative cooling systems complementing the conventional culture method.

To date, in many countries the only legally valid method for evaporative cooling system (ECS) monitoring is the culture method. However, a duration of up to 14 days and a risk of underestimation of Legionella concentrations are seen as limitations of cultivation methods. Rapid cultivation-independent methods are an important step towards a more practicable monitoring of ECS to quickly control interventions if elevated concentrations of Legionella are found. Two commercial kits for quantitative polymerase chain reaction (qPCR) and viability-qPCR (v-qPCR) were studied, comprising sample filtration and DNA extraction. Cryopreserved Legionella pneumophila were established as calibration standard with intact (ILC) and total Legionella count (TLC) determined by flow cytometry before conducting spiking experiments in commercial mineral water and artificial process water. Final assessment was carried out using real ECS samples. Recovery and robustness ranged from 86 to 108 % for qPCR with a drop to 40-60 % for v-qPCR when compared to direct extraction, possibly attributable to cell damage during sample concentration. All methods including culture did perform well regarding linearity with R2 ≥ 0.95 for most trials. Detected concentrations in comparison to spiked Legionella counts differed with culture averaging 25 ± 7 % of spiked ILC and v-qPCR being closest to spiked concentrations with 65-144 %. In comparison, qPCR was several fold above spiked TLC concentrations. For real ECS samples Legionella spp. were detected in concentrations above 103 GU/100 mL by v-qPCR in 70-92 % of samples, depending on the kit used. Most of these samples were either culture-negative or not evaluable on agar plates. This study showed that a cryopreserved bacterial standard based examination is applicable and can be used for future v-qPCR verification. For assessment of differences in results between culture and v-qPCR/qPCR in ECS samples expert knowledge about the operating mode and used analytical methods is required. Guidelines addressing this issue could be a solution.

Clinical relevance of colonization with antimicrobial-resistant bacteria (AMRB) and methicillin susceptible Staphylococcus aureus (MSSA) for mothers during pregnancy.

PURPOSE: The impact of colonization with antimicrobial-resistant bacteria (AMRB) and methicillin-sensitive Staphylococcus aureus (MSSA) of healthy pregnant women is not described in detail in Germany. In this study, we screened for MSSA and AMRB, especially for methicillin-resistant S. aureus (MRSA) as well as extended-spectrum beta-lactamase (ESBL)-producing E. coli. Potential risk factors for colonization with AMRB/MSSA and the potential effects of colonization with these on the obstetric population were investigated. METHODS: From October 2013 until December 2015 pregnant women were screened before birth for colonization with AMRB/MSSA from the mammillae, nose, perianal and vaginal area. Before birth, the expectant mother was administered a standardized interview questionnaire by a trained interviewer. Data from the hospital admission records were also included. RESULTS: Samples from 651 pregnant women were analyzed. Colonization with MSSA was detected in 14.3% (n = 93), AMRB in 3.5% [(n = 23); MRSA: n = 3/ESBL: n = 20]. Significantly more colonization of AMRB/MSSA could be detected in women who had previously given birth compared to women who were nulliparous (p < 0.05). MSSA colonization was significantly associated with self-reported respiratory diseases during pregnancy (p < 0.05), but AMRB/MSSA colonization was not statistically associated with other types of infection. CONCLUSION: Our results demonstrate a low overall rate of colonization with AMRB/MSSA, as well as a low percentage of colonized pregnant women who developed infections. Multiparous women are at higher risk for colonization with MSSA/MRSA or ESBL. Because the prevalence of AMRB/MSSA is low, this study suggests that general screening of pregnant women without risk factors is not recommended.

Evaluation of sampling locations in pregnant women and newborns for the detection of colonisation with antibiotic-resistant bacteria.

Up to now, little has been known about the prevalence and clinical relevance of colonisation of asymptomatic pregnant women with methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA) or extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli. In this two-centre cross-sectional study, we evaluated the performance and importance of screening at different times and different locations for colonisation in pregnant women and newborns. Between October 2013 and December 2015, four samples were collected from pregnant women, two from newborns at birth and three from 3-day-old newborns. Samples were screened on culturing media and were confirmed with molecular methods. MSSA was used as a surrogate for MRSA, as the two share most microbiologic characteristics and colonisation patterns. Of 763 pregnant women, 14.5% (111) were colonised with MSSA, 0.4% (3) with MRSA and 2.6% (20) with ESBL-producing E. coli. Of 658 newborns, 0.9% (10) were colonised with MSSA at birth and 13.1% (70) at 3 days old, 0.5% (3) were colonised with MRSA and 2.6% (17) with ESBL-producing E. coli. Nasal sampling identified 91.0% of MSSA-colonised pregnant women and 60.0% of newborns. In newborns, nasal and umbilical sampling at 3 days after birth discovered 84.0% of colonised cases. For ESBL-producing E. coli, the perianal region was positive in all colonised pregnant women and in 88.2% of colonised newborns. Combining nasal and perianal swabs is optimal when screening for antibiotic-resistant bacteria in pregnant women. Nasal, perianal and umbilical sample collection from 3-day-old newborns significantly increased the sensitivity compared to screening immediately after birth.

Odonto-sinusal influences.

There is a wide documentary material regarding the upper dental disease influencing the state of the maxillary sinus. This article tries to show that the relation is a two-way one.

[Anatomo-clinical considerations regarding the celiac-mesenteric junction]. / Consideratii anatomo-clinice asupra butonului celio-mezenteric.

AIM: This work points out the access ways - both the classical ones and the ones less known and used - to the origin and the first centimeters of the superior mesenteric artery (SMA), underlining the advantages and disadvantages of each of them. MATERIAL AND METHOD: Our work is based on dissection of 30 human bodies and 200 selective arteriographies. RESULTS: There are two classical ways to access the first part of SMA: inter-duodeno-pancreatic and the left thoraco-abdominal ways. CONCLUSIONS: An access way to the origin of the SMA is necessary for the revascularisation through different techniques of the visceral territory depending on the SMA, taking into account the fact that at this level we have the most frequent localization of the atheroma plaques, which leads to acute or chronic intestinal ischemia (manifest in the form of postprandial abdominal 'angor').

Conservative treatment of obesity in an academic obesity unit. Long-term outcome and drop-out.

OBJECTIVE: The aim of this study was to explore the outcome and the problems of drop-out in the treatment of obese outpatients at an academic obesity unit. DESIGN: A two-year clinical treatment evaluation. SUBJECTS: A total of 117 obese subjects, 83 women and 34 men, mean aged 50 (23-70) years, with an average body mass index (BMI) of 39.0 kg/m2 (28.8- 64.7). INTERVENTION: All treatment was based on group therapy and included behaviour modification and nutrition counselling. A team of nurses, dieticians, a physiotherapist, a psychotherapist and a physician supervised the treatment. Two programmes were used. Group 1 initially received a low-calorie diet (LCD) for seven weeks combined with the behaviour treatment programme. Group 2 was treated with the behaviour treatment programme only. All subjects were offered complementary treatment according to their medical needs. RESULTS: There was a continuous drop-out of subjects during the two-year treatment period with an overall drop-out rate of 53%. Anthropometric characteristics, medical history or reasons for drop-out had no impact on the drop-out rate. In completers the weight reduction after two years was 9.2 [+/-10.8 standard deviation (S.D.) kg. In non-completers the weight reduction of the last observed weight measurement was 4.7 (+/-7.9 S.D.) kg. After year two, the weight reduction in Group 1 was 8.8 (+/-12.2 S.D.) kg, and in Group 2 was 9.7 (+/-8.0 S.D.) kg. CONCLUSION: This study has showed the difficulties of long-term clinical treatment of obese outpatients, even in a specialised obesity clinic. The findings demonstrate that educated and experienced staff together with an extended package of treatment options is not enough to keep patients in treatment for two years. However though the drop-out rate was high, two thirds of the included subjects reduced their weight, which is a satisfactory result in a clinical setting. The drop-out rate and the reasons for dropping out could give a clue in which direction the diagnostics and analysis of the subject's individual needs in health care should be directed.

Fermentation conditions affecting the bacterial growth and exopolysaccharide production by Streptococcus thermophilus ST 111 in milk-based medium.

AIMS: To study the effect of different fermentation conditions and to model the effect of temperature and pH on different biokinetic parameters of bacterial growth and exopolysaccharides (EPS) production of Streptococcus thermophilus ST 111 in milk-based medium. METHODS AND RESULTS: The influence of temperature and pH was studied through fermentation and modelling. Fermentations under non-pH controlled conditions with S. thermophilus ST 111 indicated that the EPS production was low in milk medium, even if additional nitrogen sources were supplemented. Under pH-controlled conditions, addition of whey protein hydrolysate to the milk medium resulted in a fivefold increase of the EPS production. This medium did not contain polysaccharides interfering with EPS isolation. Primary and secondary modelling of different fermentations revealed an optimum temperature and pH of 40 degrees C and constant pH 6.2, respectively, for growth in milk medium supplemented with whey protein hydrolysate. Maximum EPS production was observed in the range of 32-42 degrees C and constant pH 5.5-6.6. Whereas growth and maximum EPS production were clearly influenced by temperature and pH, the specific EPS production was only affected by stress conditions (T = 49 degrees C). CONCLUSIONS: Addition of whey protein hydrolysate to milk medium resulted in an increased growth and EPS production of S. thermophilus ST 111 under pH-controlled conditions. A modelling approach allowed studying the influence of temperature and pH on the kinetics of both growth and EPS production. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of an appropriate milk-based medium and a combined model of temperature and pH can be of practical importance for the production of yoghurt or other fermented milks as well as for process optimization of the large-scale production of starter strains to be used for their EPS production.

Enterococcus saccharominimus sp. nov., from dairy products.

Four isolates, which were obtained from Belgian, Moroccan and Romanian dairy products, constituted a homogeneous but unidentified taxon after screening with whole-cell protein fingerprinting. Complete 16S rRNA gene sequence analysis classified representative strains in the genus Enterococcus. Highest sequence similarities of 98.6 and 98.0 % were obtained with the species Enterococcus sulfureus and Enterococcus saccharolyticus, respectively. Growth characteristics, biochemical features, tRNA intergenic length polymorphism analysis, DNA-DNA hybridization and DNA G+C contents of selected strains demonstrated that they represent a single, novel Enterococcus species. It differs phenotypically from other enterococci in characteristics commonly considered as typical of this genus: no growth in 6.5 % NaCl or 0.4 % sodium azide, and no acid production from a wide range of carbohydrates. The name Enterococcus saccharominimus sp. nov. is proposed for this novel species; the type strain (LMG 21727(T)=CCM 7220(T)) was isolated from contaminated pasteurized cow's milk.

[Correlations between the type of distribution of the branches of the inferior mesenteric artery and the aspect of the mesocolon]. / Corelatiile dintre tipurile de distributie a colateralelor arterei mezenterice inferioare si aspectul mezoului.

Emergence pattern of inferior mesenteric artery (IMA) branches leaded to description of 6 types of arterial distribution, each of them being correlated with a certain mesocolon aspect. The height of mesosigmoid depends on coalescence pattern and grade, and has a particular importance in terminal colic and retroperitoneal viscera's surgery. For each of the 6 types of distribution are described different ways of differential ligatures, for elongation and descending the colon, initially establishing the predominant collateral trunks. For practicing these ligatures we have to take count of preoperative paraclinic investigations, general and particular haemodynamic conditions and some intraoperatory parameters.

Production kinetics of acidophilin 801, a bacteriocin produced by Lactobacillus acidophilus IBB 801.

Lactobacillus acidophilus IBB 801 produces a small bacteriocin, designated acidophilin 801. Studying the relationship between growth and bacteriocin biosynthesis revealed primary metabolite kinetics of bacteriocin production with a peak activity at the end of the exponential growth phase followed by a decrease during the stationary phase. Both microbial growth and bacteriocin production was inhibited by lactic acid. Whereas volumetric bacteriocin production (activity units (AU) ml(-1)) was favoured under pH-controlled conditions, bacteriocin titres rapidly decreased because of strong adsorption of the bacteriocin molecules to the producing cells under less acidic conditions.

Purification and characterization of a bacteriocin produced by Lactobacillus acidophilus IBB 801.

Lactobacillus acidophilus IBB 801 produces a small bacteriocin, designated acidophilin 801, with an estimated molecular mass of less than 6.5 kDa. It displays a narrow inhibitory spectrum (only related lactobacilli but including the Gram-negative pathogenic bacteria Escherichia coli Row and Salmonella panama 1467) with a bactericidal activity. The antimicrobial activity of cell-free culture supernatant fluid was insensitive to catalase but sensitive to proteolytic enzymes such as trypsin, proteinase K and pronase, heat-stable (30 min at 121 degrees C), and maintained in a wide pH range. The proteinaceous compound was isolated from cell-free culture supernatant fluid and purified. Crude bacteriocin was isolated as a floating pellicle after ammonium sulphate precipitation (40% saturation) and partially purified by extraction/precipitation with chloroform/methanol (2/1, v/v). Further purification to homogeneity was performed by reversed phase Fast Performance Liquid Chromatography. The amino acid composition was determined. Amino acid sequencing revealed that the N-terminal end was blocked.

[Intermesenteric anastomoses]. / Anastomozele intermezenterice.

This work is based on dissection and the analysis of the arteriographies of two mesenteric arteries and it analyses the type of intermesenteric anastomoses and their importance in surgical practice. The morphological varieties of Riolan's arcade depends on its pillars, as well as on the eventual presence, especially in the middle part of the transversal mesocolon, of the secondary arcades; it seems that the colic wall necrosis depends less upon the location of the lesion or the level of the suture of the arcade, but more so on the stretch of the destruction of the mesocolon. The intermesenteric trunk, a second way of intermesenteric anastomoses is a embryological remaining, non-functional in normal conditions, but capable to partially or completely replace the Riolan's arcade when the later is sutured or interrupted.

[The interpretation of the telomic distribution of the pulmonary arterial trunk]. / Interpretarea distributiei telomice a trunchiului arterial pulmonar.

The analytical study of the periodicity of organic processes is of great interest as reflected by the literature. This paper aims at describing the functional implications from the viewpoint of pulmonary functional arterial circulation regularity. In the light of morphological aspects, the pressure falls along a symmetric telomic tree were followed up and compared with the experimentally obtained data.