Comparison of serum lipid compositions, lipid peroxide, alpha-tocopherol and lipoproteins in captive marine mammals (bottlenose dolphins, spotted seals and West Indian manatees) and terrestrial mammals.

Concentrations of serum lipid components, lipid peroxide (LPO) and alpha-tocopherol and electrophoretic patterns of lipoproteins in serum samples obtained from captive marine mammals and terrestrial mammals were compared. Serum concentrations of total cholesterol, free fatty acid, and phospholipid in fish-eating animals were significantly higher than those in manatees and cows. Serum LPO and alpha-tocopherol concentrations in the fish-eating animals were also significantly higher than those in manatees, cows and dogs. Different patterns of densitometric scans of low density lipoprotein (LDL) and a significantly lower percentage of LDL were demonstrated in the dolphins compared with the seals, cow and dogs. The concentration of LPO was significantly correlated with triglyceride and phospholipid concentrations in serum from the dolphins. These results suggest that triglyceride and phospholipid are susceptible to oxidative reaction in fish-eating animals. Evaluation of serum lipids, LPO and alpha-tocopherol concentrations is needed for nutritional husbandry for fish-eating animals.

Affinity of Staphylococcal a and Streptococcal G Proteins to West Indian Manatee ( Trichechus manatus manatus) Immunoglobulins.

The West Indian manatee ( Trichechus manatus manatus), a subspecies that inhabits coastal areas of Central and South America, has been listed as a vulnerable species because of the rapid decline in its population. Commercially available immunologic reagents specific for sirenians are lacking, limiting the development of sensitive immunodiagnostic assays. We observed the affinity of the microbial proteins A and G to T. m. manatus immunoglobulins. Manatee serum pools were analyzed using enzyme-linked immunosorbent assay (ELISA) to determine the affinity intensity followed by western blotting to confirm the specific binding of proteins A and G to immunoglobulins. The ELISA demonstrated maximum affinity of both proteins until the serum dilution of 1:12,800, with a similar affinity for both proteins. Because both A and G proteins exhibited affinity to manatee immunoglobulins, they can be used to develop sensitive immunodiagnostic assays for this species, contributing to manatee conversation procedures.

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 "cold stress syndrome" 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, 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 <10-50 microg/ml with an equivocal interval of 51-70 microg/ml. The reference interval for albumin:globulin ratio by plasma gel electrophoresis is 0.7-1.1. 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 0.4-2.4 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 100-400 mg/dl, a diagnostic sensitivity of 40%, and a diagnostic specificity of 95%.

From the Worm in a Bottle of Mezcal: iDNA Confirmation of a Leech Parasitizing the Antillean Manatee.

Invertebrate-derived ingested DNA (iDNA) is quickly proving to be a valuable, non-invasive tool for monitoring vertebrate species of conservation concern. Using the DNA barcoding locus, we successfully identified both the blood-feeding leech Haementeria acuecueyetzin and its blood meal-the latter is shown to be derived from the Caribbean manatee, Trichechus manatus . DNA amplification was successful despite the fact that the specimen was fixed in Mezcal (a beverage distilled from agave). We report the first confirmed case of a leech feeding on a manatee, the first record of H. acuecueyetzin for the State of Chiapas and, to our knowledge, the first case of successful DNA amplification of a biological sample fixed in Mezcal other than the caterpillar "worms" more commonly found in that beverage.

Thyroid hormone concentrations in captive and free-ranging West Indian manatees (Trichechus manatus).

Because thyroid hormones play a critical role in the regulation of metabolism, the low metabolic rates reported for manatees suggest that thyroid hormone concentrations in these animals may also be reduced. However, thyroid hormone concentrations have yet to be examined in manatees. The effects of captivity, diet and water salinity on plasma total triiodothyronine (tT(3)), total thyroxine (tT(4)) and free thyroxine (fT(4)) concentrations were assessed in adult West Indian manatees (Trichechus manatus). Free-ranging manatees exhibited significantly greater tT(4) and fT(4) concentrations than captive adults, regardless of diet, indicating that some aspect of a captive existence results in reduced T(4) concentrations. To determine whether this reduction might be related to feeding, captive adults fed on a mixed vegetable diet were switched to a strictly sea grass diet, resulting in decreased food consumption and a decrease in body mass. However, tT(4) and fT(4) concentrations were significantly elevated over initial values for 19 days. This may indicate that during periods of reduced food consumption manatees activate thyroid-hormone-promoted lipolysis to meet water and energetic requirements. Alterations in water salinity for captive animals did not induce significant changes in thyroid hormone concentrations. In spite of lower metabolic rates, thyroid hormone concentrations in captive manatees were comparable with those for other terrestrial and marine mammals, suggesting that the low metabolic rate in manatees is not attributable to reduced circulating thyroid hormone concentrations.