Quantifying, Identifying and Culturing Marine Blowhole Ciliates in Captive Beluga Whales

We quantified the abundance of ciliates (Kyaroikeus and Planilamina spp.) in the exhale of two captive beluga whales. Over a seven-week period, duplicate weekly blow samples were taken from two trained beluga whales at Mystic Aquarium, CT, USA (the 17-year-old male Juno and 38-year-old female Kela). We observed all three described blowhole species. To estimate abundance, samples were preserved in formalin, stained with DAPI, and counted using an inverted fluorescence microscope. These findings were compared to the health indicators of the whales (e.g. stress hormone levels), which were measured at the time of sampling. We had difficulty amplifying the ribosomal DNA of the ciliates, but succeeded with Kyaroikeus after first making cDNA. This is the first known sequence of a member of the family Kyaroikeidae, and we hope to use it to confirm its phylogenetic position. The 38-year-old female consistently had more ciliates than the 17-year-old male. The average abundance in the female was 1296 ciliates per exhale vs 184 for the male (p-value < 0.05). This could be due to age, size, exhale behavior, or time in captivity. In preliminary comparisons, we have found no significant correlation between the measured health indicators and the abundance of blowhole ciliates, but we are continuing this work. Using several artificial sputum formulations, we attempted to grow the ciliates in vitro, but were not successful.