An outbreak of Shewanella putrefaciens group in wild eels Anguilla anguilla L. favoured by hypoxic aquatic environments.

Microbiological analyses were conducted on wild eels from the L'Albufera Lake (Spain). A total of 174 individuals were collected in two surveys (i.e. year 2008 and autumn-winter 2014) among those caught by local fishermen into the lagoon. The prevalence of Shewanella putrefaciens group was 1.7% in 2008 and rose above 32% in 2014. It was due to an outbreak of shewanellosis that presented a morbidity rate of 64%. S. putrefaciens group strains were isolated as pure cultures from the sick eels that showed white ulcers surrounded by a reddish inflammation, damage of the mouth, extensive skin discoloration, exophthalmia, ascites and bad odour. The S. putrefaciens group was recovered from freshwater samples taken at the L'Albufera system, along autumn-winter 2015. Its counts significantly increased in freshwater parallel to hypoxia and temperature rising. Shewanellae strains were identified as S. putrefaciens and S. xiamenensis by 16S rRNA gene sequencing. These isolates recovered from sick eels or freshwater were virulent for European eel by IP challenge (LD50 106  CFU g-1 body weight). They also caused 30-38% cumulative mortality, in European eels challenged by a 2-h bath (107  CFU mL-1 ). These results suggest that shewanellosis could be transmitted through water highlighting the fact that hypoxic conditions increase this bacterium levels in water.

Multidrug-resistant (MDR) Aeromonas recovered from the metropolitan area of Valencia (Spain): diseases spectrum and prevalence in the environment.

Aeromonas infections are rare in Europe and often related to traveller's diarrhoea. A total of 185 Aeromonas isolates from river water, fish and clinical sources, recovered during a 1-year period, were used to investigate the disease spectrum and impact of multidrug-resistant (MDR) strains. They were all identified by biochemical tests and 25% of them were also identified by sequencing of the 16S rRNA gene. The minimum inhibitory concentrations (MICs) of 21 antimicrobials were determined for all isolates by broth microdilution/E-strips methods, and susceptibility was assessed according to the Clinical and Laboratory Standards Institute (CLSI). Strains pathogenicity was determined by using Swiss Webster mice as the animal model. Aeromonas diseases had an incidence of around 20 cases/million inhabitants in the metropolitan area of Valencia (Spain). Acute gastroenteritis in children with no history of travel abroad was the main pathology. These cases were related to A. caviae, A. veronii biovar sobria, A. hydrophila and A. dhakensis. A significant incidence of A. caviae in humans was found, while the other species were equally present in clinical and environmental origins. A. jandaei, A. bestiarum and A. media had mainly an environmental distribution. The prevalence of MDR Aeromonas was maximal in clinical samples, and resistance phenotypes were significantly related to this source. 7.2% of environmental Aeromonas was resistant to at least five drugs; most of them were moderately virulent for mice and, in addition, belonged to clinically significant species. The present study demonstrates a diseases spectrum similar to that reported in tropical countries, and also that pathogenic and heavily MDR Aeromonas are present in environmental reservoirs. MDR Aeromonas from any source analysed were susceptible to aztreonam, netilmicin, cefotaxime, ceftazidime, cefepime and fluoroquinolones.

Presence of viruses in wild eels Anguilla anguilla L, from the Albufera Lake (Spain).

A virological analysis was conducted on wild eels from the Albufera Lake (Spain). A total of 179 individuals at different growth stages were collected in two different surveys (2004 and 2008). Presence of anguillid herpesvirus (AngHV-1), aquabirnavirus and betanodavirus was confirmed by PCR procedures in both surveys, although the number of detections was clearly higher in 2008 (83% of the eels analysed resulted positive for virus presence). AngHV-1 was the viral agent most frequently detected, followed by aquabirnaviruses. Betanodaviruses were detected by the first time in wild eels, and although the detections were only made by nested PCR, high percentage of positives were achieved. In addition, in 2008, seven aquabirnaviruses were isolated. Phylogenetic analysis performed using partial sequences of both genomic segments of aquabirnaviruses indicated that the seven isolates could be typed as WB (genogroup I) on the basis of segment A sequences, but when segment B was used six of them clustered with C1 strain (genogroup V) and one was typed as Ab (genogroup II). These results indicate natural reassortment between different strains of aquabirnaviruses in the eels. Although betanodaviruses were not isolated in cell culture, the analysis of the sequence of the nested PCR product indicated that they clustered with SJNNV genotype. The diversity of viral agents and the high level of viral detections suggest that viral infections may play a more prominent role in the decline of the European eel than initially thought.

Is there a need to widely prescribe antibiotics in patients hospitalized with COVID-19?

BACKGROUND: Debate continues regarding the usefulness and benefits of wide prescription of antibiotics in patients hospitalized with coronavirus disease 2019 (COVID-19). METHODS: All patients hospitalized in the Infectious Diseases Department, Dijon University Hospital, Dijon, France between 27 February and 30 April 2020 with confirmed COVID-19 were included in this study. Clinical, biological and radiological data were collected, as well as treatment and outcome data. An unfavourable outcome was defined as death or transfer to the intensive care unit. Patient characteristics and outcomes were compared between patients who did and did not receive antibiotic therapy using propensity score matching. FINDINGS: Among the 222 patients included, 174 (78%) received antibiotic therapy. The univariate analysis showed that patients who received antibiotic therapy were significantly older, frailer and had more severe presentation at admission compared with patients who did not receive antibiotic therapy. Unfavourable outcomes were more common in patients who received antibiotic therapy [hazard ratio (HR) 2.94, 95% confidence interval (CI) 1.07-8.11; P = 0.04]. Multi-variate analysis and propensity score matching indicated that antibiotic therapy was not significantly associated with outcome (HR 1.612, 95% CI 0.562-4.629; P = 0.37). CONCLUSION: Antibiotics were frequently prescribed in this study and this was associated with more severe presentation at admission. However, antibiotic therapy was not associated with outcome, even after adjustment. In line with recent publications, such data support the need to streamline antibiotic therapy in patients with COVID-19.

Multiresistant waterborne pathogens isolated from water reservoirs and cooling systems.

AIMS: To determine the incidence of multiple antibiotic-resistant strains of the emergent human pathogens Legionella pneumophila, Pseudomonas aeruginosa and mesophilic Aeromonas species among those isolated from water reservoirs and industrial cooling systems. METHODS AND RESULTS: Water from four natural water reservoirs and four industrial cooling towers was sampled for 1 year period. The total heterotrophs, mesophilic Aeromonas, Pseudomonas spp. and Legionella spp. counts were performed as recommended by standard procedures, and the sensitivity of the isolates to 27 antibiotics was tested. A total of 117 Aeromonas, 60 P. aeruginosa and 15 L. pneumophila strains were isolated and identified by means of biochemical tests and DNA probes. 46.4% of Aeromonas, and 100% of P. aeruginosa isolates presented multiple resistance. Legionella pneumophila strains were generally sensitive to the drugs used. CONCLUSIONS: Antibiotic-resistant pathogenic bacteria belonging to P. aeruginosa and mesophilic Aeromonas species are common in natural aquatic environments. Thus, the risk of waterborne diseases owing to domestic and industrial uses of freshwater should be re-examined from the increase of bacterial resistance point of view. SIGNIFICANCE AND IMPACT OF THE STUDY: These data confirm the emergence of bacteria resistant to antibiotics in aquatic environments.

Ultra-High Mass Resolution MALDI Imaging Mass Spectrometry of Proteins and Metabolites in a Mouse Model of Glioblastoma.

MALDI mass spectrometry imaging is able to simultaneously determine the spatial distribution of hundreds of molecules directly from tissue sections, without labeling and without prior knowledge. Ultra-high mass resolution measurements based on Fourier-transform mass spectrometry have been utilized to resolve isobaric lipids, metabolites and tryptic peptides. Here we demonstrate the potential of 15T MALDI-FTICR MSI for molecular pathology in a mouse model of high-grade glioma. The high mass accuracy and resolving power of high field FTICR MSI enabled tumor specific proteoforms, and tumor-specific proteins with overlapping and isobaric isotopic distributions to be clearly resolved. The protein ions detected by MALDI MSI were assigned to proteins identified by region-specific microproteomics (0.8 mm2 regions isolated using laser capture microdissection) on the basis of exact mass and isotopic distribution. These label free quantitative experiments also confirmed the protein expression changes observed by MALDI MSI and revealed changes in key metabolic proteins, which were supported by in-situ metabolite MALDI MSI.

Fast and simultaneous determination of endocrine disrupting compounds by ultra-high performance liquid chromatography-tandem mass spectrometry.

A rapid and sensitive analytical method for the simultaneous determination of thirteen endocrine disruptors (five phthalates, seven parabens, and bisphenol A) in a single chromatographic run has been developed for the first time. The separation method, based on ultra-high performance liquid chromatography (UHPLC), allows the separation of all compounds (including isobaric pairs) in less than 4.1 min. The fast polarity switching mode of the triple quadrupole mass spectrometer used enables the registration of positive (phthalates) and negative (parabens and BPA) ions in the same acquisition run. A Response Surface Methodology was used for the optimization of the method. The optimum elution program starts with 0.2 min in isocratic conditions (79.8% water; 20% acetonitrile, 0.2% ammonium formate 5mM at pH 10.2), then the content of acetonitrile is linearly increased in 2 min up to 42%, and later up to 98% in 1.1 min. The analytical characteristics of the developed method were satisfactory. The method is robust and showed a linear response with determination coefficients (R(2)) higher than 0.991 in the range 5.0-2000 pg on column (or higher) for all the compounds investigated. Instrumental intra- and inter-day precision (expressed as relative standard deviation) were lower than 12% for parabens and bisphenol A, and between 5.9% and 27% for phthalates. Instrumental detection and quantification limits (iLODs and iLOQs) were in the range of medium-high femtograms (270-1300 pg on column for iLODs). Finally, the suitability of the developed method was demonstrated through its application to the analysis of commercial personal care products (shower gels) without any sample treatment, only a simple dilution, being possible to determine the simultaneous presence of phthalates, parabens, and bisphenol A in almost all the gels analyzed.

Novel strategy for the revalorization of olive (Olea europaea) residues based on the extraction of bioactive peptides.

This work proposes a new strategy for the revalorization of residual materials from table-olive and olive oil production based on the extraction of bioactive peptides. Enzymatic hydrolysates of olive seed protein isolate were prepared by treatment with five different proteases: Alcalase, Thermolysin, Neutrase, Flavourzyme and PTN. Although all hydrolysates presented antioxidant properties, Alcalase was the enzyme that yielded the hydrolysate with the highest antioxidant capacity. All hydrolysates showed antihypertensive capacity, obtaining IC50 values from 29 to 350 µg/ml. Thermolysin was the enzyme which yielded the hydrolysate with the highest ACE-inhibitory capacity. Hydrolysates were fractionated by ultrafiltration showing a high concentration of short chain peptides, which exhibited significantly higher antioxidant and antihypertensive capacities than fractions with higher molecular weights. Peptides in most active fractions were identified by LC-MS/MS, observing homologies with other recognized antioxidant and antihypertensive peptides. Finally, their antioxidant and antihypertensive capacities were evaluated after in vitro gastrointestinal digestion.

O-serogrouping and surface components of Aeromonas hydrophila and Aeromonas jandaei pathogenic for eels.

The relationship between virulence, O-serogroup, and some cell-surface features (self-pelleting [SP] and precipitation after boiling [PAB], profile of lipopolysaccharides [LPSs] and outer membrane proteins [OMPs]) was investigated in strains of the pathogenic species Aeromonas hydrophila and A. jandaei isolated from eels. Virulent strains of A. hydrophila reacted mostly with O:19 antiserum, and those of A. jandaei reacted with O:4, O:11, O:15 and O:29 antisera (Guinée and Jansen system). Regarding the PAB and LPS profiles two groups could be distinguished; (i) five PAB+ strains of serotype O:19 that possessed a homogeneous O polysaccharide side chain and (ii) thirteen PAB- strains antigenically diverse that either exhibited a heterogeneous side chain or were side chain deficient. A major 50 kDa protein was only found in the PAB+ strains, whereas major OMPs detected in PAB- strains ranged from 33 to 45 kDa irrespective of the species. Epizootic eel isolates of A. hydrophila belong to serotype O:19 and share cell-surface features with the Aeromonas highly virulent for other hosts. In contrast, epizootic A. jandaei isolates were antigenically diverse. These findings reinforce the importance of an O-serotype as an epidemiological marker in motile Aeromonas strains pathogenic for eels.

Enzyme-linked immunosorbent assay for detection of Aeromonas hydrophila serogroup O:19.

An enzyme-linked immunosorbent assay has been developed for the detection of Aeromonas hydrophila serogroup O:19 isolated from epizootics in eels. The enzyme-linked immunosorbent assay specificity was confirmed after testing A. hydrophila O:19 and non-O:19 strains from different origins, as well as other Aeromonas species and other fish pathogens such as Vibrio vulnificus biotype 2, V. furnisii, V. damsela, Yersinia ruckerii and Edwardsiella tarda. The detection limits for A. hydrophila O:19 cells were around 10(4)-10(5) cells/well. Artificially infected eels were analyzed and the immunodetection was confirmed by cultural methods. With this methodology A. hydrophila O:19 was successfully detected in infected eels and water samples. We described two subgroups within the serogroup O:19 (Guinée and Jansen system), one of them presents a 50 kDa outer membrane protein as a strong thermostable antigen which is not present in the other group.

Aeromonas allosaccharophila sp. nov., a new mesophilic member of the genus Aeromonas.

Phenotypic and genetic studies were performed on some atypical aeromonas strains of uncertain taxonomic position. 16S rRNA gene sequence analysis revealed that these strains represent a hitherto unknown genetic line within the genus Aeromonas, for which the name Aeromonas allosaccharophila sp. nov. is proposed. The type strain is CECT 4199.

Immune imbalance in cancer patients.

The immune status of the tumor-bearing patient remains poorly defined. In various solid-tumor-bearing patients, we demonstrated the absence of ADCC modifications in the patient in relapse or in evolution. These same patients presented a significant increase in immune complexes when compared with patients in remission. Furthermore, we noted a decreased NK activity, a decreased number of ARFC, corresponding to a helper T cell subpopulation, and a corollary increase in T-dependent suppressor activity. These results, on the whole, suggest an immune imbalance and that the helper cell-suppressor cell ratio should be investigated in greater depth within the context of the immune response in the cancer patient.

Effect of processing conditions on acidification properties of wheat sour doughs.

The pH, total titratable acidity (TTA) and lactic and acetic acids production have been investigated for wheat sour doughs with and without yeast addition, inoculated with two strains of Lactobacillus plantarum (B33, B39). To study the effect of flour extraction rate (0.54, 11.11 and 1.68% ash content), dough yield (DY) (160, 200 and 240 of sour dough/100 g flour), and fermentation temperature (25, 30 and 35 degrees C) a response surface regression, factor analysis and K-means clustering analysis were used. Results from factor analysis point out that the extraction rate of fluor governs TTA and acetic acid content; this factor accounts for the 53% of variability of the data. Dough yield is highly correlated with lactic acid content, explaining 27% of the total variance. Finally, temperature explains the remaining 16% of variation, but it is not related to any analytical variable. From K-means clustering analysis, flour extraction rate of 1.68% ash content leads to the highest TTA and acetic acid values, whereas DY of 240 g sour dough/100 g flour gives the greatest lactic acid content, and DY of 160 leads to the lowest levels of TTA and organic acids.

Grisorixin, an ionophorous antibiotic of the nigericin group. I. Fermentation, isolation, biological properties and structure.

Grisorixin is an ionophorous antibiotic of the nigericin group isolated from cultures of a strain of Streptomyces griseus. It shows activity against Gram-positive bacteria and fungi but is also very toxic. The isolation and purification procedures are reported. Its structure and physico-chemical properties are also described.

Allergic contact blepharoconjunctivitis due to phenylephrine eye drops.

We present two cases of sensitization to phenylephrine hydrochloride with clinical manifestation of blepharoconjunctivitis in the course of an ophthalmologic examination. Patch testing with available commercial preparations containing phenylephrine hydrochloride was positive in both patients. The other eye drops tested during the ophthalmologic examination were negative for both cases.

[Arterial blood gas analysis in acute pulmonary embolism]. / La gazométrie artérielle dans l'embolie pulmonaire aiguë.

Arterial blood gases (pH, pO2, p CO2) were studied in 100 patients with documented pulmonary embolism (Group A), confirmed by pulmonary angiography (n = 51) or scintigraphy ( n = 49). The pO2 ranged from 32 to 97 mm Hg (average 60,5 +/- 13 mm Hg). Hypoxaemia was found in 97 cases and would therefore seem to be a reliable sign of pulmonary embolism. In the three cases in which it was absent, the embolism was small. Hypoxaemia was associated with hypocapnia and alkalosis in 91 cases. However, hypoxaemia was not a specific finding; it was also present in 49 patients with suspected pulmonary embolism (Group B) in whom the diagnosis was excluded by pulmonary angiography or scintigraphy. A previous history of cardiovascular disease was found in 37 patients (76%) in this group: of the 12 remaining patients, 6 were heavy smokers and 4 were significantly obese. No correlation was found between the degree of hypoxaemia and the extent of amputation of the vascular bed on pulmonary angiography or scintigraphy. Nevertheless, a pO2 of under 50 mm Hg was always associated with a severe embolism with amputation of over 40% of the pulmonary vascular bed. A significant correlation was found between the severity of hypoxaemia and the degree of cyanosis (p less than 0,05) and ECG changes (p less than 0,01). The average pO2 was 59 +/- 12 mm Hg in patients with cardiovascular disease ( n = 21) and 55 +/- 11 mm Hg with known pulmonary disease ( n = 6). A higher average pO2 was found when these conditions were absent (61,5 +/- 13 mmHg). The difference was not statistically significant unless previous cardiac and pulmonary disease were associated (pO2 = 51 +/- 14 mm Hg, p less than 0,05).

[Permeability of venous aortocoronary bypasses one year after their implantation]. / Perméabilité des pontages veineux aorto-coronaires un an après leur implantation.

For a short-term evaluation of the patency of aortocoronary bypass vein grafts, 54 consecutive patients who underwent this operation alone were examined, irrespective of their functional state, 12.4 +/- 2.1 months after surgery. Examinations included coronary arteriography, selective opacification of the graft and ventriculography. The patients were 47 men and 7 women who had coronary arteriography for stable angina pectoris (22.4%) or a recent episode of unstable angina (59.3%) or a recent myocardial infarction (18.5%). Coronary arteriography showed one-vessel (5.5%), two-vessel (27.8%) or three-vessel (42.6) disease or stenosis of the main left coronary artery (24.1%). The mean number of distal anastomoses in multiple-vessel patients was 2.6. Peri-operative mortality was 1.04%, and the proportion of peri-operative electrocardiographic signs of necrosis was 11%. At the time of control examination, 82% of the patients had few or no symptoms, and 88% were improved by at least one functional class. 79% (97/123) of distal venous anastomoses were patent. All anastomoses were patent in 59% (22/54) and all were occluded in 7% (4/54) of the patients. Localized stenosis of the graft was found in 11% and diffuse stenosis in 4% of the cases. The patency of vein grafts was higher when the distal bed was of normal size than when it was small or poorly visualized at the initial angiography (85% vs 62%, p less than 0.01), or when the diameter of the artery bypassed was greater than 1.3 mm.(ABSTRACT TRUNCATED AT 250 WORDS)

[Long-term prognosis of spastic angina with normal or irregular coronary arteries. Apropos of 48 cases]. / Pronostic à long terme de l'angor spastique à coronaires normales ou irrégulières. A propos de 48 cas.

Forty-eight consecutive patients (37 men, 11 women, mean age 49 +/- 8 years) were followed up regularly for a mean period of 7 years. All presented with spontaneous angina, documented coronary spasm and no stenosis greater than 50 p. 100 at coronary arteriography. The first attack of pain had taken place 3 days to 9 years previously; exertion angina was also present in 47 p. 100 of the cases and severe arrhythmia in 17 p. 100. Treatment was based on calcium antagonists in doses and combinations that varied with the course of the disease. The follow-up period lasted from 16 to 122 months (mean 85 +/- 24 months). 3 patients are now known to be alive but were lost sight of after 12 to 21 months. Major cardiac complications were 1 death (2 p. 100) and 3 cases of myocardial infarction (6.6 p. 100). None of the patients died suddenly. 70 p. 100 of the remaining 41 patients became asymptomatic; angina persisted in 15 p. 100 and 14 p. 100 had episodes of severe angina but with remissions of at least one year. No predictive factor of functional deterioration or major cardiac complications could be elicited. The long-term prognosis of vasospastic angina in patients with little or no coronary stenosis is favourable, but there is a very small risk of myocardial infarction or death.