Standardized white spot syndrome virus (WSSV) inoculation procedures for intramuscular or oral routes.

In the past, strategies to control white spot syndrome virus (WSSV) were mostly tested by infectivity trials in vivo using immersion or per os inoculation of undefined WSSV infectious doses, which complicated comparisons between experiments. In this study, the reproducibility of 3 defined doses (10, 30 and 90 shrimp infectious doses 50% endpoint [SID50]) of WSSV was determined in 3 experiments using intramuscular (i.m.) or oral inoculation in specific pathogen-free (SPF) Litopenaeus vannamei. Reproducibility was determined by the time of onset of disease, cumulative mortality, and median lethal time (LT50). By i.m. route, the 3 doses induced disease between 24 and 36 h post inoculation (hpi). Cumulative mortality was 100% at 84 hpi with doses of 30 and 90 SID50 and 108 hpi with a dose of 10 SID50. The LT50 of the doses 10, 30 and 90 SID50 were 52, 51 and 49 hpi and were not significantly different (p > 0.05). Shrimp orally inoculated with 10, 30 or 90 SID50 developed disease between 24 and 36 hpi. Cumulative mortality was 100% at 108 hpi with doses of 30 and 90 SID50 and 120 hpi with a dose of 10 SID50. The LT50 of 10, 30 and 90 SID50 were 65, 57 and 50 hpi; these were significantly different from each other (p < 0.05). A dose of 30 SID50 was selected as the standard for further WSSV challenges by i.m. or oral routes. These standardized inoculation procedures may be applied to other crustacea and WSSV strains in order to achieve comparable results among experiments.

Design of laying nests in furnished cages: influence of nesting material, nest box position and seclusion.

(1) Preferences for three nesting materials and nest box positions were investigated simultaneously in two trials using a furnished cage: one with 18 individual laying hens and one with 18 groups of 5 hens. Following a habituation period in pre-test cages, every hen or group of hens was tested for 2 d: once without and once with plastic flaps at the entrance of the nest boxes. (2) Hens preferred peat and artificial turf to coated wire mesh for egg laying. (3) One nest box position was clearly preferred to both other nest boxes. The hens' choice of nest box position was influenced by the pre-test cage in which they had been habituated. (4) The presence of plastic flaps at the entrance of the nest boxes had no influence on the proportion of eggs laid on the different nesting materials or on the proportion of floor eggs. (5) Individual and group testing resulted in the same overall results despite the presence of a distinct group effect.