Avian leukosis virus (ALV) is the most common oncogenetic retrovirus that emerges spontaneously as a result of recombination between exogenous viruses, exogenous viruses and endogenous viruses, and exogenous viruses and non-homologous cellular genes. In the present study, two natural recombinant avian leukosis viruses (rALVs) (LC110515-5 and LC110803-5) carrying a subgroup C gp85 gene, a subgroup E gp37 gene, and a subgroup J 3'UTR and 3'LTR were isolated from two different lines of layer flocks, Black-bone silky fowl (BSF) and commercial layer chicken, that suffered from myeloid leukosis. Although tumours were not observed in rALV-infected individual chickens, other non-neoplastic inflammatory lesions were evident. The two rALVs were cultured on DF-1 cells and identified by PCR, immunofluorescence assay and gene sequencing. The gp85 nucleotide sequence in the two isolates displayed a high identity (>95 %) with that of the gp85 gene in ALV-C, but the identity was less than 90 % with ALV-A/B/D/E and only 51 % with ALV-J. Phylogenetic analysis of the nucleotide and amino acid sequences confirmed that the two isolates were recombinant between ALV-C, ALV-E and ALV-J. Subgroup C ALV is rarely found in field cases. This report is the first to provide evidence that ALV-C has recombined with ALV-E and ALV-J in two different chicken lines. The source and characteristics of the two rALVs and ALV-C need to be further investigated.