| Summary: | <i>Chlamydia psittaci</i>, <i>Chlamydia gallinacea,</i> and <i>Chlamydia abortus</i> are the most common <i>Chlamydia</i> spp. in chickens and have a confirmed or suggested zoonotic potential. No recent data are available on their prevalence and impact in the Belgian chicken industry or in the recreational chicken branch. Therefore, a cross-sectional epidemiological study was executed where samples were collected from both factory-farmed and backyard chickens. More specifically, pharyngeal chicken swabs were obtained from 20 chicken farms, 5 chicken abattoirs, and 38 different backyard locations and were analyzed using species-specific Polymerase Chain Reactions (PCRs) for the presence of the three avian <i>Chlamydia</i> spp. To investigate their zoonotic potential, samples were simultaneously collected from 54 backyard chicken caretakes and 37 professional chicken caretakers or abattoir employees and analyzed using species-specific PCRs as well. This study confirmed the presence of DNA of all three <i>Chlamydia</i> species in both the chicken industry and backyard settings. <i>Chlamydia psittaci</i> was the most prevalent in the industry chickens (11.0%), whereas <i>Chlamydia gallinacea</i> was the dominant species in the backyard chickens (14.5%). <i>Chlamydia abortus</i> infections were more common in the commercial chickens (9.0%) compared to the backyard chickens (2.6%). The DNA of all three species was also detected in humans (3.9% <i>Chlamydia psittaci</i>, 2.9% <i>Chlamydia gallinacea,</i> and 1.0% <i>Chlamydia abortus</i>).
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