A comparison of the use of different swab materials for optimal diagnosis of amoebic gill disease (AGD) in Atlantic salmon (Salmo salar L.).

Routine gill swabbing is a non-destructive sampling method used for the downstream qPCR detection and quantitation of the pathogen Neoparamoeba perurans, a causative agent of amoebic gill disease (AGD). Three commercially available swabs were compared aiming their application for timelier AGD diagnosis (Calgiswab® (calcium alginate fibre-tipped), Isohelix® DNA buccal and cotton wool-tipped). Calcium alginate is soluble in most sodium salts, which potentially allows the total recovery of biological material, hence a better extraction of target organisms' DNA. Thus, this study consisted of (a) an in vitro assessment involving spiking of the swabs with known amounts of amoebae and additional assessment of retrieval efficiency of amoebae from agar plates; (b) in vivo testing by swabbing of gill arches (second, third and fourth) of AGD-infected fish. Both in vitro and in vivo experiments identified an enhanced amoeba retrieval with Calgiswab® and Isohelix® swabs in comparison with cotton swabs. Additionally, the third and fourth gill arches presented significantly higher amoebic loads compared to the second gill arch. Results suggest that limiting routine gill swabbing to one or two arches, instead of all, could likely lead to reduced stress-related effects incurred by handling and sampling and a timelier diagnosis of AGD.

Study of aluminum toxicity by means of vital staining profiles in four cultivars of Phaseolus vulgaris L.

Aluminum toxicity is a very important factor limiting crop productivity on acid soils. Early effects of aluminum toxicity comprise inhibition of cell division and effects on root elongation. The plasma membrane can be the primary target of aluminum toxicity and thus, vital staining techniques could be a powerful tool in determining effects of metal stress on the plasma membrane. In this paper. we discuss the effects of Al on growth and membrane integrity by staining root tips with a mixture of fluorescein diacetate and propidium iodide. The results show a good correlation between results from growth measurement and the vital staining. From the comparison of the luminosity patterns generated by vital staining it is easy to determine Al-resistant varieties, revealing this technique as a powerful and fast method for determining tolerance to Al in different varieties.

Aluminium-induced alteration of ion homeostasis in root tip vacuoles of two maize varieties differing in Al tolerance.

Root elongation is a primary target of Al toxicity in plants. The objective of this study was to see whether Al-induced disturbance of ion homeostasis is related to the inhibition of root elongation. For this purpose, root growth rate, free cytoplasmic calcium (Ca²+) and vacuolar content of phosphate (P(i)), potassium (K+), nitrate (NO3⁻) and malate, as well as malate and citrate exudation and nitrate reductase activity were analysed in tips of two Zea mays L. varieties differing in Al resistance. Aluminium treatment affected root growth and cytoplasmic Ca²+ in the Al sensitive variety Bakero, but not in the Al tolerant variety Sikuani. However, both varieties suffered Al-induced decrease of vacuolar K+, and phosphate concentrations. Vacuolar malate concentrations were more affected by Al in Bakero than in Sikuani. Vacuolar nitrate concentrations increased upon Al exposure in both varieties. Only in Sikuani rhizosphere, pH slightly increased upon Al exposure. Our data are consistent with the hypothesis that disturbance of Ca²+ homeostasis is an early event in the Al toxicity syndrome. However, Al-induced alterations of the root tip homeostasis of major ions seem unrelated to Al-induced inhibition of root elongation.