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Campylobacter jejuni is an important food-borne pathogen. The main source of this pathogen is poultry and poultry products. Poultry farms of low biosecurity level plays major role in disseminating this pathogen. The objectives of this study were to investigate the occurrence of Campylobacter and identify potential risk factors associated with their presence in layer farms in Northern Jordan. A total of 2524 samples from chickens, litter, water and feed were collected from 35-layer farms. Samples underwent conventional and enrichment isolation methods for Campylobacter. Confirmation was done morphologically, biochemically and by PCR typing. The flock-level prevalence of C. jejuni was 40%, 37%, 20% in chicken cloacae, drinking water and litter respectively. C. jejuni was the only confirmed isolated species. None of the feed samples revealed presence of Campylobacter. The concentration of free residual chlorine was below the recommended standard levels. The risk factors were identified using modified semi-structured questionnaire. There was no significant association between evaluated risk factors and isolation status potentially reflecting small number of study farms. The prevalence rate for C. jejuni is within commonly reported range. High stocking density, short distance between farms, improper hygienic practice and low water chlorine level seems to increase occurrence rate of Campylobacter in layer farms. Educational biosecurity programs regarding C. jejuni transmission and their public health importance needs to be established.