Objective Ocular actinomycosis is an uncommon progressive infection. The study aims to investigate the predisposing factors, clinical characteristics and treatment outcomes of culture-proven cases of Actinomycotic primary canaliculitis. Methods Single-center, retrospective, interventional case series. Culture-proven cases of Actinomyces-associated primary canaliculitis diagnosed and treated between January 2017 and December 2021 at the Beijing Tongren Eye Center, Capital Medical University were identified and reviewed for ocular and systemic risk factors, clinical presentations, microbiological profile, treatment modalities and outcomes. Results Of the 201 patients diagnosed with primary canaliculitis, 22 (10.9%) were caused by Actinomyces. The mean age at onset of 22 patients (21 women and one men) was 54 years. The lower canaliculus was most commonly involved (54.6%). The most frequent symptom was mattering without epiphora (77.3%) and clinical signs were punctal regurgitation of purulent discharge (100%) and expressible concretions (95.5%). Dry eye co-existed in 77.3% of patients, whereas no obvious systemic factors were found. Among 19 cases (86.0%) of identified Actinomyces species, A. odontolyticus (43.5%) was the predominant causative microorganism. There were 50% of patients with polymicrobial infection and the most common additional bacteria isolated were Streptococcus species. Conservative therapy combining repeated canalicular expression and irrigation with susceptible topical antibiotics achieved complete resolution in 86.4% of patients. Conclusions Dry eye was identified in the vast majority of patients with Actinomycotic canaliculitis. Most cases are odontogenic in origin and the infection occurs in immunocompetent individuals. The conservative method combining canalicular expression and irrigation with topical susceptible antibiotics is recommendable as initial therapy.