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A species-discriminatory aerA TaqMan qPCR assay for rapid quantification of Aeromonas veronii in fish tissues and aquaculture water.

Aeromonas veronii is a major bacterial pathogen in freshwater aquaculture, yet rapid species-level quantification remains challenging within the genetically complex genus Aeromonas. We developed a singleplex hydrolysis-probe (TaqMan) quantitative PCR (qPCR) assay targeting an A. veronii-discriminatory region of the aerolysin gene (aerA) and validated it according to MIQE recommendations. Plasmid standards gave a linear range of 2 to 2 × 106 copies/reaction (R2 = 0.9962) with 100.5% amplification efficiency. The endpoint limit of detection was 2 copies per reaction, and 20 copies per reaction was set as the practical reporting limit based on reproducible detection and low intra- and inter-assay variation. Analytical specificity was evaluated with genomic DNA from an 18-strain panel, with reproducible amplification observed only for A. veronii. The assay was further tested in 55 fish-tissue and 11 aquaculture-water DNA extracts. NH8B-1D2 sample-process monitoring was used for matrix-level recovery correction, and tissue and water extraction blanks were undetermined. The aerA target was detected in all tested gill, stomach/intestine, spleen, kidney/head kidney, pond-water filter and Xiamen seawater filter extracts, and in 10/11 liver extracts. Median NH8B-corrected loads were highest in gill among tissues and higher in pond-water filters than in Xiamen seawater filters. A separate Vibrio harveyi inhibition-check assay indicated no obvious amplification-stage inhibition. This assay supports rapid quantification of aerA-positive A. veronii in fish and aquaculture-water matrices.

Animals

Aerosols characterization in a forested site in Québec, Canada.

Three AERAS low pressure 11 stage cascade impactors with rotatable collecting plates (LPCR) were installed at the Duchesnay forest station near Québec City and four low pressure inertial collectors (LPIC) were installed in the forest. The analysis of the aerosol deposits was performed by static fast atom beam mass spectroscopy (FABMS); SIMS and ESCA analyses were also made. The morphological characterization and the elemental volumetric composition of the aerosols were established by a scanning electron microscope equipped with an X-ray dispersive energy analyser. Micro-weighting of the aluminium substrate before and after the sampling by the AERAS impactor enabled the study of the mass-size-distribution of the aerosols according to their aerodynamic equivalent diameter. Example of the results obtained are presented in relation to air parcels' trajectories. This study has shown that it is possible to characterize the aerosols according to the origin and trajectories of the air mass with which they travel; sources along the tracks have been shown to contribute to the aerosol loading of the air masses.

Aerosols

[Intestinal absorption of digoxin in systemic sclerosis (author's transl)].

Gastro-intestinal absorption of digoxin was evaluated in 18 patients with progressive systemic sclerosis. In 8 patients a single-dose crossover study was performed after oral and intravenous administration of 0.5 mg digoxin by comparing the aera under the eight-hour plasma concentration curve. The fraction of the dose absorbed was diminished in 4 patients to less than 55%. There was a significant positive correlation between the extent of digoxin absorption and xylose renal excretion. In addition, steady state digoxin plasma levels and 24-h urinary excretion of digoxin were determined during maintenance therapy in 12 patients. In 6 patients renal excretion of digoxin was clearly less than in normal subjects during chronic dosing of the same digoxin preparation. This finding corresponded well with digoxin plasma levels below the usual therapeutic range in most of the patients. The impaired absorption of digoxin failed to correlate with the extent of the skin manifestation or the time course of the disease while there was massive oesophageal dysfunction in most of these patients. The results suggest that an inadequate therapeutic response to cardiac glycosides in patients suffering from progressive systemic sclerosis is at least partially due to impaired digoxin absorption. Similar problems could occur in therapy of the disease itself due to insufficient enteral absorption of drugs used in treatment of systemic sclerosis.

Administration, Oral

A simple technic for predicting daily maintenance dose of warfarin.

Warfarin anticoagulation data from forty-seven patients studied retrospectively was used to develop an algorithm for predicting daily maintenance dose of warfarin. A plot of prothrombin time versus cumulative warfarin loading dose was made for each of the forty-seven patients and the aera under the curve (AREA) measured from the baseline prothrombin time to a prothrombin time of 20 seconds. A linear correlation between the established daily maintenance dose of warfarin and AREA was derived regression analysis: Daily maintenance dose (mg/day) = 0.0465 x (AREA) + 1.5. The correlation was then used to predict daily maintenance dose in twenty-four patients studied prospectively. The mean prothrombin time for a seven day stabilization period after loading for all patients in the prospective study was 21.5 +/- 2.2 seconds and the seven day mean prothrombin time for each patient fell between 19 and 24 seconds. The results of the prospective study indicate that this technic is useful in the early, accurate prediction of a daily maintenance dose of warfarin.

Adult

Metabolomics and genomics reveal high diversity and concentrations of cyanopeptides during a Microcystis bloom.

Cyanobacterial blooms are an immense global problem that release complex mixtures of poorly characterized biologically active cyanopeptides into freshwater. In this study, metabolomics and genomics were used to assess the diversity and concentrations of cyanopeptides during a dense Microcystis bloom during the late summer of 2023 in Lake Champlain, a large transboundary lake situated between Canada and the United States. Despite the relatively low genetic diversity of the bloom determined by 16S rRNA metabarcoding, 151 cyanopeptides were detected by non-targeted metabolomics. This represents the most recorded cyanopeptides from a single lake plankton bloom event to date. Fifty-two cyanopeptides were previously reported and 99 represent putative new structures. Standards from the microcystin, cyanopeptolin, microginin, and anabaenopeptin groups were used to either quantify or approximate respective cyanopeptide concentrations over the sampling period. Cyanopeptolins were the most diverse (n = 68) cyanopeptides and the second most abundant, reaching 12,892 μg/L. Microginins were the second most diverse (n = 24) and reached the highest concentrations (18,262 μg/L). Anabaenopeptins were the third most diverse (n = 17) cyanopeptides, reaching 4,818 μg/L. Only 8 microcystins were detected, reaching 4,935 μg/L, where MC-LR was the dominant congener. Target cyanopeptide biosynthesis genes for microcystins (mcyE), cyanopeptolins (mcnC), anabaenopeptins (apnD), microviridins (mdnC), and aeruginosins (aerA) were also quantified using digital droplet PCR (ddPCR). The gene copy numbers for mcyE, mcnC, and apnD were highly correlated with their corresponding cyanopeptide concentrations. Overall, the studied Microcystis bloom produced a very diverse cyanopeptide mixture with high cyanopeptide concentrations including non-microcystin groups.

Microcystis

Molecular studies on the aerolysin gene of Aeromonas species and discovery of a species-specific probe for Aeromonas trota species nova.

A large group of aeromonads and other enteric microorganisms were assayed for the presence of the aerolysin gene with use of DNA-DNA hybridization. Two DNA fragments corresponding to the regulatory region (aerC) and the structural gene (aerA) were used as probes for the detection of the aerolysin gene in these strains. Sequences corresponding to the aerolysin structural gene were widespread among Aeromonas isolates. In contrast, the aerC probe was much more selective, and sequences corresponding to the aerC region were detected in only a small subset of strains. Concurrent studies using numerical taxonomy and DNA hybridization with the aerC probe on a larger set of strains led to the identification of a distinct cluster of 14 presumed atypical Aeromonas sobria strains. These strains have recently been grouped into a new species designated Aeromonas trota. Hence, the DNA fragment aerC used in the study is a species-specific gene probe for A. trota. The ability of the aerC probe to detect strains belonging to a single species suggests that there is selection pressure to maintain the clonality of this species. These results have important implications with respect to the evolution of "pathogenic profiles" among these medically important bacteria.

Aeromonas

[Digestive salmonellosis in South Vietnam].

During the period January 1969 through October 1973, rectal swabs from 13,947 diarrhea patients of all ages at infectious disease hospital and children hospital in Saigon were examined. From these specimens 610 strains of Salmonella of all group were isolated. A total of 46 serologic types were encounted. The most commonly encounted were S. paratyphi B, S. java and S. typhimurium (group B). Most of Salmonella isolated were obtained from adult gastroenteritis. We have also studied the possible relation between human and animal salmonellosis. The rodents trapped from different aera in Saigon, fish and clams collected from different markets were examined by bacteriological surveys. From these specimens, 42 strains of Salmonella were isolated for a 2,2 percent isolation rate. Our study showed that Salmonella types which were isolated frequently from animal were encounted in man.

Adolescent

[Tinea capitis: 73 cases observed in Bordeaux (author's transl)].

73 cases of tinea capitis were observed in the "Hôpital des Enfants" in Bordeaux during these last five year. 71 children under 14 years of age and only 2 adults have seen. The dermatophytic spectrum transformation noticed in other parts of Western Europe is also encountered in Bordeaux. 83 p. 100 are affected by zoo-antropophilic dermatophytes, M. canis chiefly in town, T. mentagrophytes, T. verrucosum in rural aeras. The cases are often familial, from an infected animal, but without any interhuman transmission. Only 8 out of 73 cases are due to strict antropophilic dermatophytes and are found in newly arrived immigrants from North Africa (T. violaceum, T. schönleini) of from West Africa (M. langeroni, T. ferrugineum, T. soudanense). Not a single case of scholar transmission could be observed. Cure by micronised griseofulvine per os (15 mg/kg/day) associated with local care was effective in all cases.

Adolescent

[Quantitative bone biopsy and primary hyperparathyroidism, its role in diagnosis. Apropos of 45 cases].

Iliac bone biopsy, including a quantitative study, was carried out in 38 patients with primary hyperparathyroidism (34 surgical patients, 4 with typical syndromes based on laboratory analysis). A second biopsy was carried out in 7 cases. By studying both the surface resorption of the strands of the spongy tissue and periosteocyte osteolysis (an indirect measure by assessment of the surface aera of the periosteocyte lacunae), it is possible to arrive at a histological diagnosis of hyperparathyroidism in 98 percent of cases: surface resorption is increased in 82 percent of cases; the periosteocyte lacunae are enlarged in 91 percent of cases; the two criteria together give a positive result in 44 cases out of 45 (98 percent). This morphometric study is particularly valuable in those cases in which the qualitative histological appearance is not very informative.

Adult