Development of immunity in rainbow trout (Oncorhynchus mykiss, Walbaum) to Aeromonas hydrophila after the dietary application of garlic

The development and duration of immune protection against Aeromonas hydrophila infections with garlic as immunostimulant in rainbow trout Oncorhynchus mykiss was studied. Rainbow trout fingerlings of 14 g average weight were fed with 0 g (= Control), 0.5 g and 1.0 g of garlic 100 g-1 of feed for 14 days. Physiological factors, biochemical, immunological, hematological parameters and electrolyte indices were evaluated after a further 14, 21 and 28 days before challenge with Aeromonas hydrophila. Fourteen days after the cessation of feeding with garlic, mortality rates of 12 % (relative percent survival [RPS] = 86 %) and 16 % (RPS = 80 %) were recorded in groups which received 0.5 g and 1.0 g of garlic 100 g-1 of feed, respectively, compared to 84 % mortalities in the controls. The corresponding RPS 21 days after ending the feeding regime was 75 % and 68, respectively. One week later, the RPS had dropped to 55% and 46% in the groups fed with 0.5 g and 1.0 g garlic 100 g-1 of feed, respectively

An improved 96-well turbidity assay for T4 lysozyme activity

T4 lysozyme (T4L) is an important model system for investigating the relationship between protein structure and function. Despite being extensively studied, a reliable, quantitative activity assay for T4L has not been developed. Here, we present an improved T4L turbidity assay as well as an affinity-based T4L expression and purification protocol. This assay is designed for 96-well format and utilizes conditions amenable for both T4L and other lysozymes. This protocol enables easy, efficient, and quantitative characterization of T4L variants and allows comparison between different lysozymes. Our method: • Is applicable for all lysozymes, with enhanced sensitivity for T4 lysozyme compared to other 96-well plate turbidity assays; • Utilizes standardized conditions for comparing T4 lysozyme variants and other lysozymes; and • Incorporates a simplified expression and purification protocol for T4 lysozyme