Investigation of contigs aligning to the Aphid Lethal Paralysis Virus genome, in the family Dicistroviridae, recovered a 4,125 nt contig (GenBank Q871932) spanning the RNA-dependent RNA Polymerase (RdRp) gene, the internal ribosome entry site (IRES) structure and the capsid coding region. The recovered sequence aligned with 83% nucleotide and 89% amino acid identity to the canonical ALPV genome over the RdRp gene. The two viruses shared 97% nucleotide homology along 171 nt of the IRES. The high sequence similarity between this new isolate and canonical ALPV makes it unclear whether this is a novel species or a new strain of ALPV. Regardless, ALPV has not previously been reported in association with honey bees. We propose the designation ALPV strain Brookings (after the SD county from which the virus was isolated). Specific PCR primers were designed for the Brookings strain and used to analyze additional time-course samples, resulting in detections on thirty distinct occasions, including in Mississippi, South Dakota and California. Incidence peaked in May, when 7 out of 20 hives were infected, whereas maximum abundance occurred in August albeit at a relatively low level, 4.42610 4 copies per 100 ng of RNA sample (approximately 2.21610 7 copies per bee), as compared to previously characterized honey bee viruses (Figures 3 and 4) . Frequent detection of ALPV strain Brookings throughout the time-course from multiple geographic locations suggests that this virus is not simply a ''passenger'' obtained from forage (nectar and pollen) shared with other insects. However, further investigation is required to determine whether ALPV strain Brookings is a honey bee pathogen.