Reports of a mass die-off of ∼350 elephants (Loxodonta africana) in northern Botswana over a period of two months (May–June 2020), has fuelled speculation and concern regarding the cause. Although the area in which these mortalities occurred is not protected and is considered a hotspot for human–elephant conflict and poaching, both malicious poisoning and poaching are unlikely to have played a role as other species were not affected, and elephant carcasses were found with tusks intact. In the absence of a confirmed cause we sought to identify the lines of enquiry that are most likely to lead to a definitive answer. In particular, we consider viral and bacterial agents that could precipitate species-specific mortalities on this scale, potential environmental sources of poisoning and the samples and tests that would assist in excluding/ confirming these candidate causes. Whilst it may be argued that these mortalities are unlikely to negatively impact the broader elephant population of ∼130 000 individuals in Botswana, the same cannot be said of the many vulnerable population pockets in other parts of Africa. For this reason, it is essential that the cause of the current die-off is identified as it is the only way to prevent similar losses of susceptible elephants elsewhere.