Yes, a bad inmobiliation practice does stress in animals and not only psychological stress this can change all your results in research. You must to work with personal right in work animal lab.
We evaluated effect of stress in infertility., but in relation to your question I can convey that, it is generally accepted that immobility is indicative of fearfulness, while locomotor activity is indicative of lack of fear (e.g., Boissy, 1995; Whimbey and Denenberg, 1967; Forkman et al., 2007). and also Chicken leukocyte profile changes in response to stress that it will be affected in your question. and also you can visit my page and study one of our recent paper with title of "Fear Response in Roosters Orally Exposed to
Immobilization, whether through handling or other physical restrictions, is likely to stress an animal because, if you think about it, evolutionarily speaking, being grabbed equates being predated. Therefore reactions similar to the fight or flight reflex is likely to occur, unless much time is spent habituating the animal to a particular immobilization regime. But even apparent habituation may still conceal a strss reaction. I know of a recent paper by Jacobs et al showing that the simple process of handling increases metabolic rates in lab rats. Othe researchers showed that handling of any sort affects stress experienced, and lab rats even react differently to male vs female handlers (this recent finding caused quite a stir). You can't ever discount the physiological or psychological effects of handling and immobilization in any experiment.