| Summary: | The present study aimed to show the benefits of novel lactic acid bacteria (LAB) strains isolated from the caeca of healthy chickens. These novel strains, identified as <i>Limosilactobacillus reuteri</i> and <i>Ligilactobacillus salivarius</i>, displayed high levels of lactic acid production, capability of biofilm formation, high aggregation and adhesion scores, and significant survival rates under conditions mimicking the chicken gastrointestinal tract (GIT). In addition, these novel <i>Lactobacillaceae</i> isolates were neither hemolytic nor cytotoxic. <i>In vivo</i> trials were able to establish their ability to reduce necrotic enteritis. Notably, a significant weight gain was registered, on day 10 of treatment, in the group of chickens fed with a mixture of <i>L.</i> <i>reuteri</i> ICVB416 and <i>L.</i> <i>salivarius</i> ICVB430 strains, as compared with the control group. This group has also shown a reduced number of lesions in the gut compared with other infected chicken groups. This study provides <i>in vitro</i> and <i>in vivo</i> evidence supporting the benefits of these novel <i>Lactobacillaceae</i> isolates for their use in poultry livestock as protective cultures to control the bacterial necrotic enteritis (NE) <i>Clostridium perfringens</i>.
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