Bovine herpesvirus 1 (BoHV-1) is a double-stranded enveloped DNA virus belonging to the family Alphaherpesvirinae , genus Varicellovirus . It is the causative agent of infectious bovine rhinotracheitis/infectious pustular vulvovaginitis (IBR/IPV), a worldwide disease that affects ruminants. BoHV-1 causes respiratory and reproductive symptoms, abortion, vulvovaginitis, encephalitis and fetal death (Rana and others 2011, Amoroso and others 2013). The virus is not always restricted to its natural host species. Animals such as goat, sheep, red deer and reindeer were successfully infected with BoHV-1 under experimental conditions (Thiry and others 2007). In addition, Mediterranean buffalo have been shown to be sensitive to BoHV-1 by experimental infection (Thiry and others 2007, Scicluna and others 2010), and seropositivity to BoHV-1 in water buffalo has been well documented (Peshev and Christova 2000). Nonetheless, the virus has never been identified in naturally infected buffaloes. To the authors’ knowledge, this study reports the first isolation of BoHV-1 from water buffaloes. Over the period September 2011 to December 2012, the authors received the samples of 65 buffaloes with pathologies possibly ascribable to herpesvirus: in detail, samples from 35 animals with rhinotracheitis (nasal swabs and organs), samples from 27 cases of abortion (which occurred during the third/fourth month of gestation), organs from one calf born with a hind limb deformity and euthanased when three weeks old, and organs from two calves that had died a few days (4–7) after birth. With respect to abortion, in 14 cases (which we defined complete cases) both mothers’ samples (cervical–vaginal swab and blood) and the organs of the fetus were received, in one case only the aborted fetus, and in the remaining 12 cases only cervical–vaginal swab and blood samples of the aborted buffaloes. All the samples came from animals belonging to three herds in Southern Italy. …