The microsporidium Larssoniella duplicati (Weiser, Holuša, Žižka, 2006) is a specific pathogen of the bark beetle Ips duplicatus (C.R. Sahlberg, 1836), which is a serious pest of Norway spruce (Picea abies (L.) H. Karst) in Europe. From 2011 to 2016, infection levels of L. duplicati and other pathogens in I. duplicatus populations were assessed along a gradient, ranging from areas in the north, where the beetle is native, to areas in the south, where the beetle has only recently invaded. The 21 study sites ranged in altitude from 229 to 1009 m a.s.l. We found that pathogen infection levels in I. duplicatus populations decreased from the native areas in the north to the new areas of beetle expansion in the south. We also found that pathogen level increased with altitude. The L. duplicati infection levels were not associated with the infection levels of other beetle natural enemies. The infection level decreased with the length of time of beetle establishment in an area. The infection level increased with the number of beetles trapped and dissected at a site.