Persistent long-term habitat use by Florida manatees at Fort Pierce, Florida from 1997 to 2020.

To survive cold winters, Florida manatees (Trichechus manatus latirostris) depend on artificial (i.e., power plants) and natural warm water sources such as springs and passive thermal basins. Passive thermal basins can provide critical habitat for manatees for short or extended periods of time. The Henry D. King Powerplant in Fort Pierce, Florida discharged warm water into Moore's Creek until it went offline in 1995. However, it is unknown to what degree manatees continue to occupy this area and how environmental factors influence their occurrence in the creek. To explore this, we examined the habitat use of Florida manatees in Moore's Creek after the shutdown from November 1997 to March 2020 from daily counts of manatees. In addition, we correlated local environmental data (ambient air, temperature, salinity) to assess if Moore's Creek had properties indicative of a passive thermal basin. Results indicated there was not an increase or decrease in habitat use over twenty years in the Creek. The consistent use of Moore's Creek over the study period suggests that this habitat possesses thermal and freshwater resources to support manatee occurrence long-term. These findings provide robust support for the importance of this habitat and passive thermal basins for Florida manatees.

Manatees display diel trends in acoustic activity at two microhabitats in Belize.

Many marine mammals exhibit diel trends in vocal production, which can provide information on habitat use and behavioral activity. In Belize, Antillean manatees (Trichechus manatus manatus) commonly inhabit small depressions in the substrate or deep-water coves known as "resting holes". Determining if manatees exhibit diel temporal trends in their call production rate and call types between microhabitats can provide insights into their diurnal and nocturnal activity patterns. Here, we investigate the diel vocalization patterns of wild Antillean manatees in two adjacent resting holes off of St. George's Caye, Belize. Recordings of manatees were made using a bottom-mounted hydrophone located near a reef barrier reef for nine days in July of 2017 and ten days in January of 2018. To explore if and how manatee acoustic activity differs between sites, we compared the number of calls per hour, the number of manatee positive hours, the number of tonal and atonal sounds, and the number of boats detected across sites. A total of 370 hours of acoustic recordings were analyzed resulting in the detection of 3,262 calls. There were no significant differences in the number of manatee calls produced per hour between sites. The average number of calls produced by manatees decreased over the course of several days. The proportion of tonal calls decreased with hours after sunset and increased in boat presence. These results suggest manatees in this region may exhibit different diel activity patterns which appear to be influenced by the characteristics of the environment. These findings can support ongoing conservation and management efforts to safeguard species in Belize.

Vocalizations of wild West Indian manatee vary across subspecies and geographic location.

Geographic variation in the vocal behavior of manatees has been reported but is largely unexplored. Vocalizations of wild West Indian manatees (Trichechus manatus) were recorded with hydrophones in Florida from Florida manatees (Trichechus manatus latirostris), and in Belize and Panama from Antillean manatees (Trichechus manatus manatus) to determine if calls varied between subspecies and geographic regions. Calls were visually classified into five categories: squeaks, high squeaks, squeals, squeak-squeals, and chirps. From these five categories, only three call types (squeaks, high squeaks and squeals) were observed in all three populations. Six parameters from the temporal and frequency domains were measured from the fundamental frequency of 2878 manatee vocalizations. A repeated measures PERMANOVA found significant differences for squeaks and high squeaks between each geographic location and for squeals between Belize and Florida. Almost all measured frequency and temporal parameters of manatee vocalizations differed between and within subspecies. Variables that may have influenced the variation observed may be related to sex, body size, habitat and/or other factors. Our findings provide critical information of manatee calls for wildlife monitoring and highlight the need for further study of the vocal behavior of manatees throughout their range.

Antillean manatee calves in captive rehabilitation change vocal behavior in anticipation of feeding.

Captive animals typically develop anticipatory behaviors, actions of increased frequency done in anticipation of an event such as feeding. Anticipatory behaviors can be an indicator of an animal's welfare. However, for rehabilitating animals that are expected to be reintroduced into the wild, these behaviors need to be extinguished to ensure successful release. Scheduled activities such as feeding occur daily and vocalizations could potentially be used to identify anticipatory behavior. Here, we tested the hypothesis that manatee calves modify their vocal production rate as a form of anticipatory behavior. Vocalizations of two Antillean manatee (Trichechus manatus manatus) calves were recorded for 10 min before, during, and after feeding sessions at Wildtracks, a manatee rehabilitation center in Belize. The number of calls were counted across recording sessions and three acoustic parameters were measured from calls including duration, frequency modulation, and center frequency. A repeated measures ANOVA comparing the number of calls across sessions indicated manatees produced significantly more calls before feeding sessions than during and after sessions. In addition, manatees increased the duration and lowered the frequency of calls before feeding sessions. This information can give further insight on ways to improve rehabilitation protocols and manage human interactions to increase the overall survival rate of rehabilitated manatees when released back into the wild.

Manatee calf call contour and acoustic structure varies by species and body size.

Vocal activity and signal characteristics of mammals are driven by several factors that result in both stability and plasticity over multiple time scales. All three extant species of manatee communicate with several calls that are especially important for maintaining contact between cows and calves. Determining if calf calls differ across manatee species will provide insights into the evolution of species-specific acoustic communication traits. We investigated the interspecific differences in the vocalizations of calves of Amazonian manatees (Trichechus inunguis) and the two subspecies of the West Indian manatee (T. manatus). Vocalizations of individual calves were recorded in rehabilitation centers in Brazil, Puerto Rico, the United States, and Mexico. The acoustic structure of calls produced by manatee calves varied between species and with body size. Amazonian manatee calves produced shorter calls with multiple notes at higher frequency while West Indian calves produced modulated calls that were lower in frequency and longer in duration. Smaller West Indian calves produced frequency modulated, hill-shaped calls that flattened with an increase in body length. Our results provide evidence for divergence in the ontogeny of vocalizations across T. manatus and T. inunguis and suggest variation in body size contributed to the evolution of differences in the characteristics of their calls.

The use of non-transplant biologics in solid organ transplant recipients: A practical review for the frontline clinician.

Biologics have become the forefront of medicine for management of autoimmune conditions, leading to improved quality of life. Many autoimmune conditions occur in solid organ transplant (SOT) recipients and persist following transplant. However, the use of biologics in this patient population is not well studied, and questions arise related to risk of infection and adjustments to induction and maintenance immunosuppression. Guidelines have been published highlighting management strategies of biologics around the time of elective surgical procedures, but this is not always feasible in urgent situations, especially with deceased donor transplantation. The aim of this review is to summarize the current literature regarding the use of these agents in solid organ transplant recipients, and specifically address induction and maintenance immunosuppression, as well as the need for alternative infective prevention strategies to create a practical reference for the frontline clinician, when faced with this complex clinical scenario.

First characterization of vocalizations and passive acoustic monitoring of the vulnerable African manatee (Trichechus senegalensis).

Even among the understudied sirenians, African manatees (Trichechus senegalensis) are a poorly understood, elusive, and vulnerable species that is difficult to detect. We used passive acoustic monitoring in the first effort to acoustically detect African manatees and provide the first characterization of their vocalizations. Within two 3-day periods at Lake Ossa, Cameroon, at least 3367 individual African manatee vocalizations were detected such that most vocalizations were detected in the middle of the night and at dusk. Call characteristics such as fundamental frequency, duration, harmonics, subharmonics, and emphasized band were characterized for 289 high-quality tonal vocalizations with a minimum signal-to-noise ratio of 4.5 dB. African manatee vocalizations have a fundamental frequency of 4.65 ± 0.700 kHz (mean ± SD), duration of 0.181 ± 0.069 s, 97% contained harmonics, 21% contained subharmonics, and 27% had an emphasized band other than the fundamental frequency. Altogether, the structure of African manatee vocalizations is similar to other manatee species. We suggest utilizing passive acoustic monitoring to fill in the gaps in understanding the distribution and biology of African manatees.

Classification of Florida manatee (Trichechus manatus latirostris) vocalizations.

The vocal repertoire for the Florida manatee is quantitatively categorized from a sample of 1114 calls recorded from 3 different manatee habitats in Florida. First, manatee vocalizations were categorized into five call categories based on visual inspection of spectrograms and following descriptions provided in previous studies. Second, based on measurements of 17 acoustic parameters, the subjective classification scheme was validated using classification and regression trees (CARTs) and model-based cluster analysis paired with silhouette coefficients. CART analysis revealed that these five broad call categories can be successfully distinguished based on correct classification scores of 41.6%-62.5%. Silhouette coefficients determined that the manatee vocal repertoire is highly intergraded. This study supports and expands upon existing subjective categorization schemes by providing a quantifiable methodology for describing the Florida manatees' vocal repertoire. These findings contribute to the increasing number of studies suggesting many animal species vocal repertoires contain graded call types.

The Antillean manatee produces broadband vocalizations with ultrasonic frequencies.

Antillean manatees produce vocalizations reported to be important for communication, but their vocal behavior throughout their geographic range is poorly understood. A SoundTrap recorder (sample rates: 288/576 kHz) was deployed in Belize to record vocalizations of wild manatees in a seagrass channel and of a young rehabilitated and released manatee in a shallow lagoon. Spectral analysis revealed broadband vocalizations with frequencies up to 150 kHz and a high proportion of calls with ultrasonic components. Ultrasonic frequency components appear prevalent in their vocal repertoire and may be important to manatee communication.

Design and characterization of a direct ELISA for the detection and quantification of leucomalachite green.

Malachite green (MG), a member of the N-methylated triphenylmethane class of dyes, has long been used to control fungal and protozoan infections in fish. MG is easily absorbed by fish during waterborne exposure and is rapidly metabolized into leucomalachite green (LMG), which is known for its long residence time in edible fish tissue. This paper describes the development of an enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of LMG in fish tissue. This development includes a simple and versatile method for the conversion of LMG to monodesmethyl-LMG, which is then conjugated to bovine serum albumin (BSA) to produce an immunogenic material. Rabbit polyclonal antibodies are generated against this immunogen, purified and used to develop a direct competitive enzyme-linked immunosorbent assay (ELISA) for the screening and quantification of LMG in fish tissue. The assay performed well, with a limit of detection (LOD) and limit of quantification (LOQ) of 0.1 and 0.3 ng g(-1) of fish tissue, respectively. The average extraction efficiency from a matrix of tilapia fillets was approximately 73% and the day-to-day reproducibility for these extractions in the assay was between 5 and 10%.