The use of animal cell cultures as tools for studying the microsporidia of insect and mammals is briefly reviewed, along with an in depth review of the literature on using fish cell cultures to study the microsporidia of fish. Fish cell cultures have been used less often but have had some successes. Very short-term primary cultures have been used to show how microsporidia spores can modulate the activities of phagocytes. The most successful microsporidia/fish cell culture system has been relatively long-term primary cultures of salmonid leukocytes for culturing Nucleospora salmonis. Surprisingly, this system can also support the development of Enterocytozoon bienusi, which is of mammalian origin. Some modest success has been achieved in growing Pseudoloma neurophilia on several different fish cell lines. The eel cell line, EP-1, appears to be the only published example of any fish cell line being permanently infected with microsporidia, in this case Heterosporis anguillarum. These cell culture approaches promise to be valuable in understanding and treating microsporidia infections in fish, which are increasingly of economic importance.

Escape rooms are physical games set in fictional settings where participants work together to find hidden objects, complete tasks, and solve puzzles to escape the room in a limited amount of time. Escape rooms are problem-based, collaborative, and require interaction with physical aspects of the game. Portable escape boxes are games developed using escape room concepts but can be taken into the classroom. Instead of escaping a room, students work together to complete tasks to get deeper into the escape box. The examination of pathophysiology case studies is well suited for the use of escape boxes since they are inherently problem-based. Using escape boxes is a natural progression to further problem and team-based learning in the use of case studies in pathophysiology and medical education. This article explores the development of an escape box, presents the challenges and narrative of the game and discusses the lessons learned during the process.

Anncaliia algerae is an aquatic microsporidium that most commonly infects mosquitoes but can be grown on the rabbit kidney cell line, RK-13. Spores were purified from RK-13 cultures and added to cell lines from warm water fish and from an insect. The cell lines were GFSK-S1 and GFB3C-W1 from goldfish skin and brain respectively, ZEB2J from zebrafish embryos, FHMT-W1 from fathead minnow testis, and Sf9 from ovaries of a fall armyworm moth. All cultures were maintained at 27°C. Infection was judged to have taken place by the appearance of sporonts and/or spores in cells and occurred in all cell lines. Spores were also isolated from ZEB2J cultures and used to successfully infect new cultures of ZEB2J, RK-13 and Sf9. These results suggest that cells of a wide range of vertebrates support A. algerae growth in vitro and fish cells can produce spores infectious to cells of mammals, fish, and insects.