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Background  Histomonosis is a severe re-emerging disease of poultry caused by  Histomonas meleagridis , a protozoan parasite which survives in the environment  via  the cecal worm  Heterakis gallinarum . Following infection, the parasites reside in the ceca and are excreted  via  host feces. In the present work, male birds of conventional broiler (Ross 308, R), layer (Lohmann Brown Plus, LB) and a dual-purpose (Lohmann Dual, LD) chicken line were infected with 250 embryonated eggs of  Ascaridia galli  and  Heterakis gallinarum , respectively, with the latter nematode harboring  Histomonas meleagridis , to investigate a co-infection of nematodes with the protozoan parasite in different host lines.    Methods  In weekly intervals, from 2 to 9 weeks post infection (wpi), individual fecal samples ( n  = 234) from the chickens were collected to quantify the excretion of  H. meleagridis  by real-time PCR and to determine the number of nematode eggs per gram (EPG) in order to elucidate excretion dynamics of the flagellate and the nematodes. This was further investigated by indirect detection using plasma samples of the birds to detect antibodies specific for  H. meleagridis  and worms by ELISA. The infection with  H. meleagridis  was confirmed by histopathology and immunohistochemistry to detect the flagellate in the cecum of representing birds.    Results  The excretion of  H. meleagridis  could already be observed from the 2nd wpi in some birds and increased to 100% in the last week of the experiment in all groups independent of the genetic line. This increase could be confirmed by ELISA, even though the number of excreted  H. meleagridis  per bird was generally low. Overall, histomonads were detected in 60% to 78% of birds with temporary differences between the different genetic lines, which also showed variations in the EPG and worm burden of both nematodes.    Conclusions  The infection with  H. gallinarum  eggs contaminated with  H. meleagridis  led to a permanent excretion of the flagellate in host feces. Differences in the excretion of  H. meleagridis  in the feces of genetically different host lines occurred intermittently. The excretion of the protozoan or its vector  H. gallinarum  was mostly exclusive, showing a negative interaction between the two parasites in the same host.    Graphic abstractSupplementary Information  The online version contains supplementary material available at 10.1186/s13071-021-04823-1.

Excretion of Histomonas meleagridis following experimental co-infection of distinct chicken lines with Heterakis gallinarum and Ascaridia galli