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R. K. Bonde

Publications and source records attributed to R. K. Bonde.

At least 19 recordsLinked to original sources

Low genetic variation and evidence of limited dispersal in the regionally important Belize manatee

The Antillean subspecies of the West Indian manatee Trichechus manatus is found throughout Central and South America and the Caribbean. Because of severe hunting pressure during the 17th through 19th centuries, only small populations of the once widespread aquatic mammal remain. Fortunately, protections in Belize reduced hunting in the 1930s and allowed the country's manatee population to become the largest breeding population in the Wider Caribbean. However, increasing and emerging anthropogenic threats such as coastal development, pollution, watercraft collision and net entanglement represent challenges to this ecologically important population. To inform conservation and management decisions, a comprehensive molecular investigation of the genetic diversity, relatedness and population structure of the Belize manatee population was conducted using mitochondrial and microsatellite DNA. Compared with other mammal populations, a low degree of genetic diversity was detected ( H E =0.455; N A =3.4), corresponding to the small population size and long-term exploitation. Manatees from the Belize City Cayes and Southern Lagoon system were genetically different, with microsatellite and mitochondrial F ST values of 0.029 and 0.078, respectively ( P ≤0.05). This, along with the distinct habitats and threats, indicates that separate protection of these two groups would best preserve the region's diversity. The Belize population and Florida subspecies appear to be unrelated with microsatellite and mitochondrial F ST values of 0.141 and 0.63, respectively ( P ≤0.001), supporting the subspecies designations and suggesting low vagility throughout the northern Caribbean habitat. Further monitoring and protection may allow an increase in the Belize manatee genetic diversity and population size. A large and expanding Belize population could potentially assist in the recovery of other threatened or functionally extinct Central American Antillean manatee populations.

Animal Conservation

Validation of a serum immunoassay to measure progesterone and diagnose pregnancy in the West Indian manatee (Trichechus manatus)

The objective was to validate a high-sensitivity chemiluminescent assay of serum progesterone concentrations for pregnancy diagnosis in manatees. Assay analytical sensitivity was 0.1 ng/mL, with mean intra- and inter-assay coefficients of variation of 9.7 and 9.2%, respectively, and accuracy had a mean adjusted R2 of 0.98. Methods comparison (relative to Siemen's Coat-A-Count RIA) demonstrated r = 0.98, Deming regression slope of 0.95, and an intercept of 0.01. Based on ROC analysis, a progesterone concentration ???0.4 ng/mL was indicative of pregnancy. Assay results were not significantly altered by two freeze-thaw cycles of samples. Characteristic progesterone concentrations during pregnancy were Months 1-4 (1.7-4.7 ng/mL), 5-8 (???1.0 ng/mL), and 10 and 11 (0.3-0.5 ng/mL), whereas two late-pregnant females with impending abortion had progesterone concentrations of 0.1 ng/mL. Among pregnant females, maximum progesterone concentrations occurred in autumn (3.9 ?? 1.8 ng/mL), and were greater during all seasons than concentrations in non-pregnant females (0.1-0.2 ng/mL). Progesterone concentrations were also significantly higher in pregnant females than in non-pregnant females and males. This highly sensitive, specific, and diagnostic assay will be valuable for monitoring pregnancy and abortion in manatees. ?? 2008 Elsevier Inc.

Theriogenology

Polymorphic microsatellite DNA markers for the Florida manatee (Trichechus manatus latirostris)

Florida manatees (Trichechus manatus latirostris) are marine mammals that inhabit the coastal waters and rivers of the southeastern USA, primarily Florida. Previous studies have shown that Florida manatees have low mitochondrial DNA variability, suggesting that nuclear DNA loci are necessary for discriminatory analyses. Here we report 10 polymorphic microsatellite loci with an average of 4.2 alleles per locus, and average heterozygosity of 50.1%. These loci have been developed for use in population studies, parentage assignment, and individual identification. ?? 2007 Blackwell Publishing Ltd.

Molecular Ecology Notes

Chromosome painting among Proboscidea, Hyracoidea and Sirenia: Support for Paenungulata (Afrotheria, Mammalia) but not Tethytheria

Despite marked improvements in the interpretation of systematic relationships within Eutheria, particular nodes, including Paenungulata (Hyracoidea, Sirenia and Proboscidea), remain ambiguous. The combination of a rapid radiation, a deep divergence and an extensive morphological diversification has resulted in a limited phylogenetic signal confounding resolution within this clade both at the morphological and nucleotide levels. Cross-species chromosome painting was used to delineate regions of homology between Loxodonta africana (2n = 56), Procavia capensis (2n=54), Trichechus manatus latirostris (2n = 48) and an outgroup taxon, the aardvark (Orycteropus afer, 2n = 20). Changes specific to each lineage were identified and although the presence of a minimum of 11 synapomorphies confirmed the monophyly of Paenungulata, no change characterizing intrapaenungulate relationships was evident. The reconstruction of an ancestral paenungulate karyotype and the estimation of rates of chromosomal evolution indicate a reduced rate of genomic repatterning following the paenungulate radiation. In comparison to data available for other mammalian taxa, the paenungulate rate of chromosomal evolution is slow to moderate. As a consequence, the absence of a chromosomal character uniting two paenungulates (at the level of resolution characterized in this study) may be due to a reduced rate of chromosomal change relative to the length of time separating successive divergence events. ?? 2007 The Royal Society.

Proceedings of the Royal Society B: Biological Sci

An epizootic of Florida manatees associated with a dinoflagellate bloom

Over a 10-wk period in early 1982, 39 Florida manatees ( Trichechus manatus latirostris ) were found dead in the lower Caloosahatchee River and nearby waters of southwestern Florida. Two were killed by boats. The remainder showed no evidence of trauma. Lesions indicative of infectious agents were not identified, and bacteriological and contaminant residue findings were unremarkable. Nonspecific lesions of congestion and hemorrhage were identified in brain tissue. Numerous reports were also received of manatee morbidity. Some distressed manatees showed no biochemical lesions in clinical analyses of blood samples and recovered quickly. Timing of manatee illnesses coincided with fish and double-crested cormorant ( Phalacrocorax auritus ) mortality and morbidity. A widespread bloom of the dinoflagellate red tide organism ( Gymnodinium breve ) also coincided with these incidents. G. breve produces potent neurotoxins (brevetoxins). Circumstantial evidence links these events, and possible routes of exposure may include ingestion of filter-feeding ascidians. Ecological conditions that magnified the extent of the epizootic included an early dispersal of manatees into the area from a nearby winter aggregation site and unusually high salinities that facilitated the inshore spread of the red tide bloom. Management responses to future episodes of red tide in manatee areas are suggested.

Marine Mammal Science

Comparison of methods used to diagnose generalized inflammatory disease in manatees (Trichechus manatus latirostris)

Manatees ( Trichechus manatus latirostris ) are afflicted with inflammatory and infectious disease secondary to human interaction, such as boat strike and entanglement, as well as &ldquo;cold stress syndrome&rdquo; and pneumonia. White-blood-cell count and fever, primary indicators of systemic inflammation in most species, are insensitive in diagnosing inflammatory disease in manatees. Acute phase-response proteins, such as haptoglobin and serum amyloid A, have proven to be sensitive measures of inflammation/infection in domestic large animal species. This study assessed diagnosis of generalized inflammatory disease by different methods including total white-blood-cell count, albumin: globulin ratio, gel electrophoresis analysis, C-reactive protein, alpha 1 acid glycoprotein, haptoglobin, fibrinogen, and serum amyloid A. Samples were collected from 71 apparently healthy and 27 diseased animals during diagnostic medical examination. Serum amyloid A, measured by ELISA, followed by albumin:globulin ratio, measured by plasma gel electrophoresis, were most sensitive in diagnosing inflammatory disease, with diagnostic sensitivity and specificity of approximately 90%. The reference interval for serum amyloid A is <10&ndash;50 &mu;g/ml with an equivocal interval of 51&ndash;70 &mu;g/ml. The reference interval for albumin:globulin ratio by plasma gel electrophoresis is 0.7&ndash;1.1. Albumin: globulin ratio, calculated using biochemical techniques, was not accurate due to overestimation of albumin by bromcresol green dye-binding methodology. Albumin:globulin ratio, measured by serum gel electrophoresis, has a low sensitivity of 15% due to the lack of fibrinogen in the sample. Haptoglobin, measured by hemoglobin titration, had a reference interval of 0.4&ndash;2.4 mg/ml, a diagnostic sensitivity of 60%, and a diagnostic specificity of 93%. The haptoglobin assay is significantly affected by hemolysis. Fibrinogen, measured by heat precipitation, has a reference interval of 100&ndash;400 mg/dl, a diagnostic sensitivity of 40%, and a diagnostic specificity of 95%.

Journal of Zoo and Wildlife Medicine

Electrocardiography in two subspecies of manatee (Trichechus manatus latirostris and Trichechus manatus manatus)

Electrocardiographic (ECG) measurements were recorded in two subspecies of awake, apparently healthy, wild manatees (Trichechus manatus latirostris and T. m. manatus) undergoing routine field examinations in Florida and Belize. Six unsedated juveniles (dependent and independent calves) and 6 adults were restrained in ventral recumbency for ECG measurements. Six lead ECGs were recorded for all manatees and the following parameters were determined: heart rate and rhythm; P, QRS, and T wave morphology, amplitude, and duration; and mean electrical axis (MEA). Statistical differences using a t-test for equality of means were determined. No statistical difference was seen based on sex or subspecies of manatees in the above measured criteria. Statistical differences existed in heart rate (P = 0.047), P wave duration (P = 0.019), PR interval (P = 0.025), and MEA (P = 0.021) between adult manatees and calves. Our findings revealed normal sinus rhythms, no detectable arrhythmias, prolonged PR and QT intervals, prolonged P wave duration, and small R wave amplitude as compared with cetacea and other marine mammals. This paper documents the techniques for and baseline recordings of ECGs in juvenile and adult free-living manatees. It also demonstrates that continual assessment of cardiac electrical activity in the awake manatee can be completed and can be used to aid veterinarians and biologists in routine health assessment, during procedures, and in detecting the presence of cardiac disease or dysfunction.

Journal of Zoo and Wildlife Medicine

Molecular identification of a papilloma virus from cutaneous lesions of captive and free-ranging Florida manatees

Cutaneous papillomatous lesions were biopsied from three captive Florida manatees ( Trichechus manatus latirostris ) at Homosassa Springs State Wildlife Park (HSSWP), Homosassa, Florida, USA, and from six free-ranging Florida manatees from Crystal and Homosassa rivers, Florida. Total DNA extracted from these lesions was assayed for the presence of papilloma virus genomes using the polymerase chain reaction (PCR) with primers that target the L1 capsid protein gene. The amplification generated DNA fragments 458 base pairs in length that encompassed a highly conserved domain within the L1 capsid protein and translated into identical polypeptides of 152 amino acids, suggesting the involvement of a single papilloma virus genotype. Multiple amino acid sequence and phylogenetic analyses of the L1 fragment indicated that the Florida manatee papilloma virus is a unique and quite distinct papillomavirus from other known papilloma viruses. The emergence of this new pathogen raises concerns about its potential impact on the already endangered Florida manatee.

Journal of Wildlife Diseases

Seasonal movements, migratory behavior, and site fidelity of West Indian manatees along the Atlantic coast of the United States

The West Indian manatee ( Trichechus manatus ) is endangered by human activities throughout its range, including the U.S. Atlantic coast where habitat degradation from coastal development and manatee deaths from watercraft collisions have been particularly severe. We radio-tagged and tracked 78 manatees along the east coast of Florida and Georgia over a 12-year period (1986-1998). Our goals were to characterize the seasonal movements, migratory behavior, and site fidelity of manatees in this region in order to provide information for the development of effective conservation strategies. Most study animals were tracked remotely with the Argos satellite system, which yielded a mean (SD) of 3.7 (1.6) locations per day; all were regularly tracked in the field using conventional radiotelemetry methods. The combined data collection effort yielded >93,000 locations over nearly 32,000 tag-days. The median duration of tracking was 8.3 months per individual, but numerous manatees were tracked over multiple years (max = 6.8 years). Most manatees migrated seasonally over large distances between a northerly warm-season range and a southerly winter range (median one-way distance = 280 km, max = 830 km), but 12% of individuals were resident in a relatively small area (<50 km) year-round. The movements of one adult male spanned >2,300 km of coastline between southeastern Florida and Rhode Island. No study animals journeyed to the Gulf coast of Florida. Regions heavily utilized by tagged manatees included: Fernandina Beach, FL to Brunswick, GA in the warm season; northern Biscayne Bay to Port Everglades, FL in the winter; and central coastal Florida, especially the Banana River and northern Indian River lagoons, in all seasons. Daily travel rate, defined as the distance between successive mean daily locations, averaged 2.5 km (SD = 1.7), but this varied with season, migratory pattern, and sex. Adult males traveled a significantly greater distance per day than did adult females for most of the warm season, which corresponded closely with the principal period of breeding activity, but there was no difference between the sexes in daily travel rate during the winter. The timing of seasonal migrations differed markedly between geographic regions. Most long-distance movements in the southern half of the study area occurred between November and March in response to changing temperatures, whereas most migrations in the northern region took place during the warmer, non-winter months. Manatees left their warm-season range in central Florida in response to cold fronts that dropped water temperatures by an average of 2.0??C over the 24-hr period preceding departure. Water temperature at departure from the warm-season range averaged 19??C, but varied among individuals (16-22??C) and was not related to body size or female reproductive status. The presence of industrial warm-water effluents permitted many manatees to overwinter north of their historic winter range, and for some migrants this delayed autumn migrations and facilitated earlier spring migrations. Southward autumn and northward spring migrations lasted an average of 10 and 15 days at mean rates of 33.5 (SD = 7.6) and 27.3 (SD = 10.5) km/day, respectively. The highest rate of travel during migration was 87 km/day (3.6 km/hr) during winter. Manatees overwintering in southeastern Florida often traveled north during mild weather - sometimes reaching their warm-season range - only to return south again with the next major cold front. Manatees were consistent in their seasonal movement patterns across years and showed strong fidelity, to warm-season and winter ranges. Within a season, individuals usually occupied only 1 or 2 core use areas that encompassed about 90% of daily locations. Most manatees returned faithfully to the same seasonal ranges year after year (median distance between range centers was <5 km between years). Seasonal movements of 4 immature manatees tracked as calves with their mothers

Florida, Georgia

Manatee mortality in Puerto Rico

The most pressing problem in the effective management of the West Indian manatee (Trichechus manatus) in Puerto Rico is mortality due to human activities. We assessed 90 cases of manatee strandings in Puerto Rico based on historical data and a coordinated carcass salvage effort from 1990 through 1995. We determined patterns of mortality, including type of event, condition of carcasses, spatial and temporal distribution, gender, size/age class, and the cause of death. The spatial distribution of stranding events was not uniform, with the north, northeast, and south coasts having the highest numbers. Six clusters representing the highest incidence included the areas of Fajardo and Ceiba, Bahia de Jobos, Toa Baja, Guayanilla, Cabo Rojo, and Rio Grande to Luquillo. The number of reported cases has increased at an average rate of 9.6%/yr since 1990. The seasonality of stranding events showed a bimodal pattern, from February through April and in August and September. Most identified causes of death were due to human interaction, especially captures and watercraft collisions. Natural causes usually involved dependent calves. From 1990 through 1995, most deaths were attributed to watercraft collisions. A reduction in anthropogenic mortality of this endangered species can be accomplished only through education and a proactive management and conservation plan that includes law enforcement, mortality assessment, scientific research, rescue and rehabilitation, and inter- and intraagency cooperation.

Environmental Management

Radio-tracking manatees from land and space: tag design, implementation, and lessons learned from long-term study

West Indian manatees (Trichechus manatus) were tracked along the Atlantic coast of Florida and Georgia (N = 83 manatees, n = 439 tag deployments, 1986-1996) and in eastern Puerto Rico (N = 8, n = 43, 1992-1996) using conventional and satellite-based radio-telemetry systems. A floating radio-tag, attached by a flexible tether to a padded belt around the base of the tail, enabled us to track manatees in saltwater environments. The tag incorporated VHF (very high frequency) and ultrasonic transmitters for field tracking and tag recovery, and an Argos satellite-monitored transmitter for remote tracking. We located each animal in the field about twice per week, received more than 60 000 good-quality Argos locations, and recovered tags in over 90% of deployments. The tag was designed to detach from the belt when entangled to prevent injury or drowning, and this often led to premature termination of tracking bouts. We had considerable success, however, in retagging belted manatees without recapture (97% of 392 retagging events). Most individuals were radio-tagged more than once (median = 3.0, maximum = 43) for a median total duration of 7.5 months (maximum = 6.8 yr). Data obtained through Argos have been valuable in addressing questions relating to long-distance movements, site fidelity, and identification of high-use areas. Fine-scale analyses of manatee habitat use and movements may require restricting the data set to the highest location quality or developing new analytical techniques to incorporate locational error. Field tracking provided useful ancillary data on life-history parameters, but sample sizes were small and survival estimates imprecise. Modification of the existing tag design to include Global Positioning System (GPS) functionality, with its finer spatial and temporal resolution, will offer new opportunities to address critical research and management problems facing this endangered species.

Marine Technology Society Journal