Age effects on blood gas, spirometry, airway reactivity, and bronchoalveolar lavage fluid cytology in clinically healthy horses.

BACKGROUND: Despite the increasing number of geriatric horses attended by veterinarians, there is a lack of understanding of aging-related changes on the respiratory system of horses. OBJECTIVE: To identify aging-related changes on the respiratory function and bronchoalveolar lavage fluid (BALF) cytology of horses. ANIMALS: Fifteen healthy young adult (2-11 years) and 16 healthy aged (≥20 years) horses. METHODS: The respiratory system was examined by measurement of arterial blood gases (ABG), use of respiratory inductive plethysmography (RIP) for assessment of breathing pattern and ventilatory parameters, histamine bronchoprovocation, and BALF cytology. RESULTS: No significant differences were detected with regard to values obtained by ABG or bronchoprovocation of young adult and aged healthy horses. In aged horses, there were significant differences in mean ± SD of the following parameters when compared to young horses: prolonged expiratory time (Te) measured by RIP (3.9 ± 1.5 s versus 3.0 ± 0.6 s), decreased percentage of alveolar macrophages (40.6 ± 11.3% versus 53.5 ± 9.6%), and increased percentage of lymphocytes (53.4 ± 9.5% versus 43.9 ± 11.0%). No correlations between airway reactivity and ventilatory parameters, ABG, or BALF cytology were found in this asymptomatic population. CONCLUSIONS: These results suggest that aging does not cause changes in the results obtained by ABG, most RIP-derived variables, and bronchoprovocation in the horse. A decreased percentage of macrophage and an increased percentage of lymphocytes in the BALF cytology may be expected in the asymptomatic geriatric horse and may be a result of aging.

Adhesion of water stressed Helicobacter pylori to abiotic surfaces.

AIM: The main aim of this work was to study and compare the adhesion of water exposed Helicobacter pylori to six different substrata and correlate any changes in morphology, physiology, ability to form aggregates and cultivability when in the planktonic or in the sessile phase. METHODS AND RESULTS: The number of total cells adhered for different water exposure times and modifications in the cell shape were evaluated using epifluorescence and scanning electron microscopy, and physiology assessed using Syto9 and propidium iodide (PI) cellular uptake. All abiotic surfaces were rapidly colonized by H. pylori, and colonization appeared to reach a steady state after 96 h with levels ranging from 2.3 x 10(6) to 3.6 x 10(6) total cells cm(-2). Cell morphology was largely dependent on the support material, with spiral bacteria, associated with the infectious form of H. pylori, subsisting in a higher percentage on nonpolymeric substrata. Also, sessile bacteria were generally able to retain the spiral shape for longer when compared with planktonic bacteria, which became coccoid more quickly. The formation of large aggregates, which may act as a protection mechanism against the negative impact of the stressful external environmental conditions, was mostly observed on the surface of copper coupons. However, Syto9 and PI staining indicates that most of H. pylori attached to copper or SS304 have a compromised cell membrane after only 48 h. Cultivability methods were only able to detect the bacteria up to the 2 h exposure-time and at very low levels (up to 500 CFU cm(-2)). CONCLUSIONS: The fact that the pathogen is able to adhere, retain the spiral morphology for longer and form large aggregates when attached to different plumbing materials appeared to point to pipe materials in general, and copper plumbing in particular, as a possible reservoir of virulent H. pylori in water distribution systems. However, the Syto9/PI staining results and cultivability methods indicate that the attached H. pylori cells quickly enter in a nonviable physiological state. SIGNIFICANCE AND IMPACT OF THE STUDY: This represents the first study of H. pylori behaviour in water-exposed abiotic surfaces. It suggests that co-aggregation with the autochthonous heterotrophic consortia present in water is necessary for a longer survival of the pathogen in biofilms associated to drinking water systems.

Nutrient shock and incubation atmosphere influence recovery of culturable Helicobacter pylori from water.

Three different media-Columbia agar, Wilkins-Chalgren agar, and Helicobacter pylori special peptone agar-were prepared in a diluted version and compared to the standard medium formulation in order to study a possible nutrient shock effect observed when recovering H. pylori from water by counting the number of CFU. This same parameter was subsequently used to evaluate the influence of the incubation atmosphere by using a modular atmosphere-controlled system to provide different atmospheres and by employing an established gas generation kit as a control. Both a low nutrient content of the media and a rapidly achieved microaerophilic incubation atmosphere proved to increase the numbers of environment-stressed H. pylori organisms recovered. An atmosphere of 5% CO(2), 5% O(2), and 3% H(2) is recommended, although other atmospheres with a low oxygen concentration are also acceptable. Besides highlighting and assessing the importance of several factors in the culturability of H. pylori, this paper demonstrates the potential ability to develop an optimized technique for recovery of this pathogen from water.

Monitoring cell detachment by surfactants in a parallel plate flow chamber.

The efficacy of the surfactants SDS and CTAB in detaching P. fluorescens from glass surface was evaluated in a parallel plate flow chamber. This device enables "in situ" determinations of cells detachment following the application of surfactants under well controlled hydrodynamic conditions. The results showed that SDS was able to remove almost all adhering bacteria in a short period of time, whereas CTAB did not promote much cell desorption. On the contrary, this surfactant increased the adhesion strength between cells and glass. Both surfactants promoted different alterations of cell surface properties, which explain their dissimilar effectiveness as cleansing agents.

The effect of clay particles on the activity of suspended autotrophic nitrifying bacteria and on the performance of an air-lift reactor.

Clay minerals have some properties, namely a high surface area and the ability of ion exchange that may exert some effects on microbial systems. It is often difficult to know the way the clay is exerting its influence and whether its presence improves a given metabolic process. The present work concerns the study of the effect of the addition of powdered kaolin to autotrophic nitrification systems, and includes the study of the effects of the particles on the activity of a suspended nitrifying bacteria consortium and on the performance of an air-lift biofilm reactor used for tertiary nitrification. Concerning the suspended culture, kaolin particles produced stimulation on the specific endogenous and exogenous respiration rates of the bacteria, probably due to a nutritional effect supplied by the clay. This effect was more pronounced for the ammonia oxidation rates, although nitrite oxidation was also enhanced but to a lesser extent. In respect to the presence of kaolin particles in the air-lift reactor, the results obtained indicate that the clay particles become incorporated in the biofilm pellets, but do not change significantly their thickness or their shape. However, nitrate production decreased when the concentration of particles increased. The low adsorption of ammonia by the kaolin indicated that the clay particles embedded in the biofilm did not probably retain the ions. Although it was not proved, precipitation of salts may have occurred.