Blood values of juvenile northern elephant seals (Mirounga angustirostris) obtained using a portable clinical analyzer

Background — Sick, injured, or orphaned juvenile northern elephant seals ( Mirounga angustisrostris ) treated at rehabilitation centers frequently present with abnormalities in blood sodium, potassium, chloride, BUN, and glucose concentrations, and HCT. These abnormalities could be detected rapidly using a portable blood analyzer, but the results with this analysis method do not necessarily equate with those obtained using other techniques.
Objective — The objective of this study was to better assess the clinical relevance of blood values obtained from a portable analyzer and to compare the results with values obtained using more common methods of analysis.
Methods — Heparinized whole blood samples were collected from 20 rehabilitated juvenile northern elephant seals. A portable clinical analyzer (i-STAT, i-STAT Corp, East Windsor, NJ, USA) was used to establish baseline values. Serum biochemical values were obtained using an automated chemistry analyzer (Olympus AU5200, Olympus America, Melville, NY, USA). HCT was determined using EDTA whole blood and a cell counter.
Results — Using the portable analyzer, mean (minimum-maximum) values were obtained for sodium, 143 (132–146) mmol/L; potassium, 4.4 (3.9–5.8) mmol/L; chloride, 106 (101–109) mmol/L; BUN, 1.8 (1.1–2.4) mmol/L; glucose, 7.55 (5.99–8.49) mmol/L; and HCT, 0.55 (0.52–0.61) L/L. Average differences between methods were small for potassium (-0.45 mmol/L), BUN (0.1 mmol/L), and HCT (0.037 L/L) but were large for sodium (-6.8 mmol/L), chloride (-6.4 mmol/L), and glucose (-0.56 mmol/L).
Conclusions — These results suggest that the i-STAT portable analyzer could be useful for clinically assessing juvenile elephant seals. However, when making medical decisions, the clinician should be aware of differences associated with various analyzers and sample types.     

Causes of erroneous white blood cell counts and differentials in clinically healthy young northern elephant seals (Mirounga angustirostris).

From 1993 to 1995, approximately 10% of the clinically healthy northern elephant seals (Mirounga angustirostris) at The Marine Mammal Center in California exhibited a large unexplained increase in their white blood cell (WBC) count. In these animals, WBC counts ranged from 28,780 to 125,000/mm3, with a mean of 50,087/mm3. Significant correlations between the leukocytosis and weight gain and day of admittance were identified, but no correlation existed between leukocytosis and general state of health, sex, length of stay, or diet. Bone marrow contamination of blood samples, erroneous automated leukocyte counts, and leukogram changes consistent with subclinical inflammation were the major factors contributing to the elevated WBC counts in these apparently clinically healthy animals.     

The effect of rehabilitation of northern elephant seals (Mirounga angustirostris) on antimicrobial resistance of commensal Escherichia coli

The aim of this study was to determine if antimicrobial drug use increases resistance of commensal gastrointensinal Escherichia coli of wild northern elephant seals (Mirounga angustirostris) treated in rehabilitation, and, if so, identify the risk factors involved. Minimum inhibitory concentration (MIC) levels of twelve antimicrobial drugs were determined for 289 E. coli isolates from 99 seals sampled at admission and 277 isolates obtained at release from rehabilitation using broth microdilution. Prevalence of E. coli antimicrobial resistance, MIC(50), MIC(90), and clustering of MIC values were determined for seals and the data were analyzed using Fisher's exact test, ordinal logistic regression and negative binomial regression. At release from rehabilitation 77.8% of the seals had antimicrobial resistant E. coli compared to 38.4% of the seals at admission. The MIC(90) for amoxicillin-clavulanic acid, chloramphenicol, enrofloxacin, ticarcillin-clavulanic acid, and trimethoprim-saulfamethoxazole were at levels considered to be sensitive at admission but they increased to levels of resistance at release. E. coli were grouped into four clusters by their MIC values, with increasing levels of resistance going from Cluster 1 to 4. A primary risk factor associated with the probability of a seal having E. coli in Clusters 3 and 4 was time in rehabilitation, regardless of whether the animal received treatment with antimicrobial drugs, suggesting nosocomial infection. The results of this study provide evidence that increased levels of hygiene and appropriate use of antimicrobial therapy might be important in the rehabilitation of wild animals to prevent rise in the prevalence of antimicrobial resistant bacteria.     

Amoxicillin pharmacokinetics in harbor seals (Phoca vitulina) and northern elephant seals (Mirounga angustirostris) following single dose intravenous administration: implications for interspecific dose scaling

The pharmacokinetics of sodium amoxicillin after a single intravenous dose of 20 mg/kg were determined in ten harbor seals (Phoca vitulina) and ten northern elephant seals (Mirounga angustirostris). The seals ranged in age from 1 to 6 months and the mean weights were 11.7 kg (range, 9.5-18.5 kg) for harbor seals and 47.1 kg (range, 39.5-61.4 kg) for elephant seals. The median half-life of amoxicillin (quartiles) in harbor seals, 1.5 (1.0-3.1) h. was not statistically different from that of elephant seals, 2.0 (1.4-3.8) h, nor were the differences between the terminal elimination rate constants between the two species. The only statistically significant differences between species were for area-under-the-curve (AUC), and total systemic clearance. The lack of statistical significance for differences in the volume of distribution at steady-state (Vss) may have been due to minor differences in the time frame of data collection and dose administered between the two groups. A true physiologic difference in drug handling, possibly related to renal perfusion or tubal secretory efficiency could affect amoxicillin kinetics in these species, and longer administration intervals may be appropriate for elephant seals as compared to harbor seals when administering multiple dose amoxicillin therapy at 20 mg/kg.     

Urate nephrolithiasis in a northern elephant seal (Mirounga angustirostris) and a California sea lion (Zalophus californianus).

Nephrolithiasis has rarely been reported in marine mammals. During 2004 and 2005, two cases of nephrolithiasis were diagnosed during routine necropsy examination, one in a northern elephant seal (Mirounga angustirostris) and one in a California sea lion (Zalophus californianus). Nephroliths were found throughout both kidneys during necropsy examination, varying in size from 1-10 mm in diameter in the northern elephant seal and from 1-15 mm in diameter in the California sea lion. Necropsy and histopathology revealed nephroliths in association with renal pelvic dilation and pyelonephritis in both animals. In addition, hydronephrosis was noted in the sea lion. Nephroliths were composed of uric acid and ammonium urate in the northern elephant seal and of ammonium urate in the California sea lion. The underlying disease leading to nephrolith formation was not determined; however, it is hypothesized that unknown metabolic derangements due to morphologic or physiologic differences may have played a role. This is the first report of urate nephrolithiasis in the California sea lion and northern elephant seal.     

Systemic mycosis caused by Scedosporium apiospermum in a stranded northern elephant seal (Mirounga angustirostris) undergoing rehabilitation.

A recently weaned, stranded, male northern elephant seal (Mirounga angustirostris) pup that had been undergoing rehabilitation was found severely obtunded with hyponatremia, hypokalemia, hypochloremia, and hypophosphatemia after a history of intermittent regurgitation. The animal was euthanatized, and gross postmortem findings included multifocal abscessation affecting brain, spleen, kidney, muscle, and subcutaneous tissue. Scedosporium apiospermum and mixed bacteria were cultured from brain, kidney, and subcutaneous tissue. Histopathologic examination revealed multiple fungal granulomas of variable size in the kidneys, brain, liver, and skeletal muscle. This is the first report of S. apiospermum infection associated with lesions in a marine mammal.     

Antagonism of some cyclohexamine-based drug combinations used for chemical restraint of southern elephant seals (Mirounga leonina)

SUMMARY This study examined the use of 4 antagonists of chemical restraint in mature female southern elephant seals (Mirounga leonina) that were restrained with ketamine and diazepam, ketamine and xylazine, or tiletamine and zolazepam. The antagonists were: 4-aminopyridine, yohimbine, doxapram and sarmazenil. The effects of the antagonists on the animal's time to first movement forward and recovery, heart rate, respiratory rate and venous blood gas and pH values, and level of chemical restraint were recorded. Sarmazenil (1.0 mg/kg) and doxapram (5.0 mg/kg) partially antagonised 50:1 ketamine: diazepam (ketamine = 3.0 mg/kg, diazepam = 0.06 mg/kg) and tiletamine and zolazepam (tiletamine = 0.5 mg/kg, zolazepam = 0.5 mg/kg). However, the rapid recovery after low doses of anaesthetics means that antagonism is usually unnecessary, and it may increase the likelihood of shaking. Routine antagonism of ketamine and xylazine (ketamine = 3.0 mg/kg, xylazine = 0.5 mg/kg) is more useful given its usually delayed recovery time and potential for thermoregulatory problems. For this purpose yohimbine (0.06 mg/kg) offered advantages over doxapram in giving a smoother recovery with less aggression. 4-aminopyridine (0.2 mg/kg) prolonged chemical restraint by 100:1 ketamine: diazepam (ketamine = 3.0 mg/kg, diazepam = 0.03 mg/kg) and ketamine and xylazine, and should be contraindicated. Doxapram (5.0 mg/kg) was the most useful general antagonist for all groups of drugs but shaking was seen and a lower dose is recommended.     

Effects of age,size and condition of elephant seals (Mirounga leonina) on their intravenous anaesthesia with tiletamine and zolazepam

Southern elephant seals (Mirounga leonina) were caught as part of a long-term demographic study on Macquarie Island. Over 18 months, 1033 seals were caught by hand and anaesthetised intravenously with a 1:1 mixture of tiletamine and zolazepam. Assessments were made of the effects of variations in the body condition and age at capture of the seals on the characteristics of their anaesthesia, including induction time and weighted recovery time. The size and condition of the seals were assessed by morphometric and ultrasound measurements. Weighted recovery times decreased as the body condition and age of the seals increased, but there were no residual effects of sex. There were no fatalities, and no periods of apnoea longer than five minutes were recorded. In individual seals there was a significant increase in weighted recovery time with successive captures.     

Ultrastructural,Sensory and Functional Anatomy of the Northern Elephant Seal (Mirounga angustirostris) Facial Vibrissae

Vibrissae (whiskers) play a key role in underwater orientation in foraging phocids through vibrotactile sensation processing. Our aim was to evaluate the structure of northern elephant seal (NES) vibrissae by means of light (LM) and transmission electron microscopy (TEM), in order to elucidate their function. Vibrissal follicles were processed using standardized laboratory methods and LM/TEM techniques. Individual follicular axonal numbers were counted and axonal diameter measured and averaged. NES have mystacial, rhinal, supraorbital and labial vibrissae. The vibrissal follicles are histologically subdivided into a ring, upper and lower cavernous sinuses (LCS). Each vibrissa is innervated by the deep vibrissal nerve. The average number of axons per large mystacial vibrissa is 1804 (±123), rhinal 985 (±241), supraorbital 1,064 (±204) and 374 (±65) in labial vibrissa. The entire vibrissal system carries an estimated 148 573 axons, and mystacial vibrissae alone have 125 323 axons. Axonal conduction velocity for each vibrissal type is 55.26 m/s for labial, 56.58 m/s for rhinal and 35.88 m/s for mystacial vibrissae. TEM and LM revealed a plethora of mechanoreceptors within the vibrissal follicles: Merkel cell‐neurite complexes, lanceolate and pilo‐Ruffini end organs. A vast number of sensory axons projecting from the entire vibrissal system indicate that the vibrissal sensory area takes up a large proportion of phocids’ somatosensory cortex. In conclusion, NES has highly sensitive and finely tuned vibrotactile vibrissal sense organs.     

Serum chemistry values for free-ranging ringed seals (Pusa hispida) in Svalbard

BACKGROUND: Diseases and abnormal physiologic conditions can alter the concentrations of enzymes, metabolites, minerals, and hormones in the blood of animals. The ringed seal (Pusa hispida) has been selected as a key species for environmental monitoring, but information on disease and health parameters for this species is scarce. OBJECTIVES: The aim of the study reported here was to obtain serum chemistry reference intervals for free-ranging ringed seals in Svalbard, and then to evaluate serum chemistry values in relation to age, body condition, and sex. METHODS: Blood samples were collected after death from ringed seals in Wijdefjorden and Billefjorden, Svalbard (2002-2003; n = 75). Serum was analyzed for 24 selected serum chemistry parameters (enzymes, protein, metabolites, minerals, and cortisol). RESULTS: Compared with younger or older animals, seals between 7 and 16 years of age had larger variations in the activities of alanine transaminase, aspartate transaminase, lactate dehydrogenase, and creatine kinase (CK). Animals classified as having low body condition status had more variation in the serum activity of these enzymes, compared with that in animals with higher condition scores. Serum cortisol concentration was higher in young animals (1-5 years) than in older animals. Serum CK activity was higher in males than in females. CONCLUSION: The data reported here may be useful in monitoring the health of ringed seals and for tracking the impact of environmental changes in the Arctic.     

Reference ranges for hematologic and serum biochemical values in llamas (Lama glama)

Hematologic and serum biochemical values were determined in 174 llamas of all age groups and both sexes from ranches in California and Nevada. Compared with hematologic values for horses and cattle, llama erythrocytes were more numerous (10.1 to 17.3 x 10(6)/microliters), but the PCV was lower (25 to 45%) because the smaller elliptical cells pack tighter. The mean corpuscular volume was half that of horses and cattle (22 to 29.5 fl). The mean corpuscular hemoglobin concentration was higher (38.9 to 46.2 g/dl), and the mean corpuscular hemoglobin slightly lower (9.6 to 12.6 pg). Most serum biochemical values were similar to those of cattle and horses, with the exception of triiodothyronine (48 to 468 ng/dl) and thyroxin (9.8 to 30 micrograms/dl), which are up to 10 times higher than values for other domestic species.     

Hematology and chemistry reference values for free-ranging harbor seals (Phoca vitulina) and the effects of hemolysis on chemistry values of captive harbor seals.

Most reported laboratory reference values for harbor seals (Phoca vitulina) are derived from captive seals, or stranded seals that have recovered from disease in marine mammal centers. This study established hematology and serum chemistry reference values for free-ranging harbor seals, using methods and that are current and readily available, and determined the effects of hemolysis on serum chemistry values of captive harbor seals. Blood samples were collected for hematologic and serum chemistry measurements from 14 clinically normal, adult male and female harbor seals and two juvenile harbor seals (approximate age 6 mo) captured in saltwater sloughs and estuaries near Moss Landing, California, USA. Values for amylase, globulin, and differential leukocyte count, not previously reported, were determined. In general, hematology and chemistry values in adults were similar to those reported for free-ranging and captive harbor seals, except for glucose, urea nitrogen, and lactate dehydrogenase (LDH) values, which were higher than those reported previously. Red blood cell counts in the two juveniles were higher than in adults and in young harbor seals studied previously. To determine the effects of hemolysis on serum chemistry values, two intensities of hemolysis were generated experimentally in blood collected from 11 harbor seals recovering from injuries or stranding at the Marine Mammal Center (Sausalito, California 94965, USA). Moderate hemolysis (++, 1 g/L hemoglobin, red-tinged) significantly increased LDH activity, whereas severe hemolysis ( , 2 g/L hemoglobin, cherry red) significantly increased total protein, albumin, calculated globulin, LDH, and total bilirubin and significantly decreased creatinine. The effects of hemolysis must be considered when chemistry results of harbor seals are interpreted.