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Diminishing <i>Campylobacter</i> prevalence in poultry flocks has proven to be extremely challenging. To date, efficacious control measures to reduce <i>Campylobacter</i> prevalence are still missing. A potential approach to control <i>Campylobacter</i> in modern poultry productions is to occupy its niche in the mucosal layer by administering live intestinal microbiota from adult chickens to dayold-chicks (competitive exclusion (CE)). Therefore, this in vivo study investigates the efficacy of a complex CE culture to reduce <i>Campylobacter</i> (<i>C.</i>) <i>jejuni</i> colonization in broiler chickens. For this purpose, the complex CE culture was applied twice: once by spray application to day-old chicks immediately after hatching (on the 1st day of life) and subsequently by an additional application via drinking water on the 25th day of life. We observed a consistent and statistically significant reduction of <i>C. jejuni</i> counts in cloacal swabs throughout the entire fattening period. At the end of the trial after necropsy (at 33 days of age), <i>C. jejuni</i> cecal counts also showed a statistically significant decrease of 1 log₁₀ MPN/g compared to the control group. Likewise, colon counts were reduced by 2.0 log₁₀ MPN/g. These results suggest that CE cultures can be considered a practically relevant control strategy to reduce <i>C. jejuni</i> colonization in broiler chickens on poultry farms.