Investigating an increase in Florida manatee mortalities using a proteomic approach

Two large-scale Florida manatee (Trichechus manatus latirostris) mortality episodes were reported on separate coasts of Florida in 2013. The east coast mortality episode was associated with an unknown etiology in the Indian River Lagoon (IRL). The west coast mortality episode was attributed to a persistent Karenia brevis algal bloom or ‘red tide’ centered in Southwest Florida. Manatees from the IRL also had signs of cold stress. To investigate these two mortality episodes, two proteomic experiments were performed, using two-dimensional difference in gel electrophoresis (2D-DIGE) and isobaric tags for relative and absolute quantification (iTRAQ) LC–MS/MS. Manatees from the IRL displayed increased levels of several proteins in their serum samples compared to controls, including kininogen-1 isoform 1, alpha-1-microglobulin/bikunen precursor, histidine-rich glycoprotein, properdin, and complement C4-A isoform 1. In the red tide group, the following proteins were increased: ceruloplasmin, pyruvate kinase isozymes M1/M2 isoform 3, angiotensinogen, complement C4-A isoform 1, and complement C3. These proteins are associated with acute-phase response, amyloid formation and accumulation, copper and iron homeostasis, the complement cascade pathway, and other important cellular functions. The increased level of complement C4 protein observed in the red tide group was confirmed through the use of Western Blot.

Nematode epibionts on skin of the Florida manatee, Trichechus manatus latirostris

A survey for the presence of nematodes on the skin of the native Florida manatee, Trichechus manatus latirostris from Crystal River, Florida was conducted during annual manatee health assessments. A putative isolate of Cutidiplogaster manati (Diplogastridae) and two other nematodes belonging to the same family were recovered from mid-dorsal tail skin-scrapings from all sampled winter-collected healthy wild adult manatees during two successive years (2018–2019). Qualitative abundance estimates of these three species of diplogastrid nematodes suggest that an average wild Florida manatee adult might possess between 30,000 and 120,000 nematodes on its tail dorsum and that the entire body dorsum including the tail might possess 160,000–640,000 nematodes in roughly equal ratios. Attempts to culture these nematodes on a variety of different culture media were unsuccessful but examination of the mouth (stomatal) morphology suggests specialized feeding on microbes such as diatoms or predation on other nematodes. No skin lesions were observed during the 2018–2019 samplings suggesting that under normal conditions these nematodes are highly specialized free-living epibionts of the skin that are tightly bound to this niche and horizontally transferred between individual manatees in an analogous fashion to human skin mites (Demodex folliculorum and D. brevis). Molecular phylogenetic inferences using sequences of near full length SSU and D2–D3 expansion segments of LSU rRNA genes revealed a putative new morphospecies in Cutidiplogaster sister to C. manati that was monophyletic with several named Mononchoides species, and another putative new morphospecies that formed a clade with several undescribed species similar in appearance to Mononchoides as well as Tylopharyx, Eudiplogasterium, Paroigolaimella and Sachsia.

Quantifying sublethal Florida manatee-watercraft interactions by examining scars on manatee carcasses

A leading human-related threat to the Florida manatee Trichechus manatus latirostris is collisions with watercraft, which account for 20-25% of reported mortalities. Quantitative threat assessments do not include information on all known manatee-watercraft interactions. These interactions often result in sublethal wounding, usually leaving multiple fresh external wounds in a variety of patterns. These wounds then resolve into well-healed scars. We characterized and quantified watercraft-related scar patterns (1 pattern = 1 strike event) on 2935 nonperinatal carcasses (>150 cm total length) that were recovered from 2007 through 2016 to compare the number of patterns by life stage, sex, and population region and across years. We used generalized linear mixed models to examine the effects of several factors on the probability carcasses having scars and on the number of scar patterns per carcass. The models indicated that approximately 96% of adults, approximately 70% of subadults, and approximately 34% of calves had watercraft-related scars. The raw data showed that 1 in 4 adults had been hit 10 or more times; 5 adult carcasses bore evidence of 40 or more strikes. On average, adult females had more scar patterns than did adult males. Manatees on Florida’s west coast had more scar patterns than did those on the east coast, while carcasses from the less populated Everglades had significantly fewer scar patterns than did those from the rest of the state. These results improve our understanding of the extent of sublethal injury of the Florida manatee caused by boat strikes.