Clinical biochemistry in healthy manatees (Trichechus manatus latirostris).

Florida manatees (Trichechus manatus latirostris) are endangered aquatic mammals living in coastal and riverine waterways of Florida and adjacent states. Serum or plasma biochemical analyses are important tools in evaluating the health of free-ranging and captive manatees. The purpose of this study was to measure diagnostically important analytes in the plasma of healthy manatees and to determine whether there was significant variation with respect to location (free-ranging versus captive), age class (small calves, large calves, subadults, adults), and gender. No significant differences in plasma sodium, potassium, bilirubin, glucose, alanine aminotransferase, or creatine kinase were found among these classes of animals. Compared to free-ranging manatees, captive animals had significantly lower mean concentrations of plasma chloride, phosphate, magnesium, triglycerides, anion gap, and lactate. Captive manatees had significantly higher mean values of total CO2, calcium, urea, creatinine, alkaline phosphatase, gamma-glutamyltransferase, total protein, albumin, and albumin/globulin ratio than did free-ranging animals. Differences in the environments of these two groups, including diet, temperature, salinity, and stress, might account for some of these results. The higher plasma lactate and anion gap concentrations and lower total CO2 concentrations of free-ranging manatees were probably due to greater exertion during capture, but the lack of elevated plasma creatine kinase activity relative to captive animals indicates that there was no serious muscle injury associated with capture. Plasma phosphate decreased and total globulins increased with age. Plasma cholesterol and triglyceride concentrations were highest in small calves. Plasma aspartate aminotransferase was higher in large calves than in adults and subadults, and the albumin/ globulin ratio was higher in subadults than in adults. Plasma total CO2 was higher and chloride was slightly lower in females than in males.     

Occurrence of Cryptosporidium spp. in Antillean manatees (Trichechus manatus) and Amazonian manatees (Trichechus inunguis) from Brazil

Infections by Cryptosporidium spp. in aquatic mammals is a major concern due to the possibility of the waterborne transmission of oocysts. The aim of the present study was to report the occurrence of Cryptosporidium spp. in Antillean manatees (Trichechus manatus) and Amazonian manatees (Trichechus inunguis) from Brazil. Fecal samples were collected and processed using Kinyoun's method. Positive samples were also submitted to the direct immunofluorescence test. The results revealed the presence of Cryptosporidium spp. oocysts in 12.5% (17/136) of the material obtained from the Antillean manatees and in 4.3% (05/115) of the samples from the Amazonian manatees. Cryptosporidium spp. infection was more prevalent in captive animals than in free-ranging specimens.     

Electrocardiography in two subspecies of manatee (Trichechus manatus latirostris and T. m. 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.     

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 α-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 α-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 α-tocopherol concentrations is needed for nutritional husbandry for fish-eating animals.     

Production of nitric oxide by peripheral blood mononuclear cells from the Florida manatee, Trichechus manatus latirostris

Florida manatees (Trichechus manatus latirostris) are exposed to many conditions in their habitat that may adversely impact health and impair immune function in this endangered species. In an effort to increase the current knowledge base regarding the manatee immune system, the production of an important reactive nitrogen intermediate, nitric oxide (NO), by manatee peripheral blood mononuclear cells (PBMC) was investigated. PBMC from healthy captive manatees were stimulated with LPS, IFN-gamma, or TNF-alpha, either alone or in various combinations, with NO production assessed after 24, 48, 72, and 96 h of culture. NO production in response to LPS stimulation was significantly greater after 48, 72, or 96 h of culture compared to NO production after 24h of culture. A specific inhibitor of inducible nitric oxide synthase (iNOS), L-NIL (L-N(6)-(1-iminoethyl)lysine), significantly decreased NO production by LPS-stimulated manatee PBMC. Manatee specific oligonucleotide primers for iNOS were designed to measure expression of relative amounts of mRNA in LPS-stimulated manatee PBMC from captive manatees. NO production by PBMC from manatees exposed to red tide toxins was analyzed, with significantly greater NO production by both unstimulated and LPS stimulated PBMC from red tide exposed compared with healthy captive or cold-stress manatees. Free-ranging manatees produced significantly lower amounts of nitric oxide compared to either captive or red tide rescued manatees. Results presented in this paper contribute to the current understanding of manatee immune function and represent the first report of nitric oxide production in the immune system of a marine mammal.     

Serum chemistry of free-ranging white whales (Delphinapterus leucas) in Svalbard

BACKGROUND: Abnormal physiological conditions and diseases can change the concentrations of enzymes, metabolites, and minerals in the body. Serum chemistry information may thus be indicative of a specific disease; interpretation of such information requires knowledge of serum chemistry reference intervals from a seemingly healthy population of the species. OBJECTIVE: The aim of this study was to obtain serum chemistry reference intervals for a population of white whales. METHODS: Blood samples were collected from 21 free-ranging white whales (beluga; Delphinapterus leucas). The whales were live-captured in nets during 1996-2001 in Storfjorden, Van Mijenfjorden, and Van Keulenfjorden (Svalbard, Norway). While the whales were briefly physically restrained, blood was collected from the caudal vein into vacuum tubes without anticoagulant. The blood was left to clot for 4-6 hours before serum was obtained by centrifugation. The serum samples were then kept at -20 degrees C until analysis. Enzymes (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase [ALP], creatine kinase, lactate dehydrogenase [LDH], amylase, lipase), metabolites (urea, creatinine, bilirubin, cholesterol, triglycerides, nonesterified fatty acids, glucose), and minerals (calcium, phosphate, magnesium, sodium, potassium, chloride) were analyzed in an Advia 1650 System (Bayer, Tarrytown, NY, USA). Cortisol was analyzed in an Immulite One system (Diagnostic Products Corporation, Los Angeles, CA, USA). The major blood proteins (albumin and globulins) were separated by gel electrophoresis in a Beckman Paragon electrophoresis system (Beckman Coulter, Inc., Fullerton, CA, USA). RESULTS: Serum values for all analytes were reported as median and range, and reference intervals were calculated as 10-90th percentiles. Activities of ALP and LDH and cortisol concentration were higher, and protein and bilirubin concentrations were lower compared with those previously reported for white whales from Canada; remaining results were strikingly similar in these 2 white whale populations. CONCLUSIONS: These data provide valuable serum chemistry reference intervals for future health assessments of white whales in Svalbard and other white whale populations, as well as captive individuals.     

Mycobacteriosis in two captive Florida manatees (Trichechus manatus latirostris).

Two male Florida manatees (Trichechus manatus latirostris) died at the marine aquarium in Inagi City, Tokyo, Japan. Acid-fast bacteria were demonstrated in tuberculoid nodules in the lungs from both manatees. Mycobacterium marinum and M. fortuitum were isolated from one manatee; M. marinum and M. kansasii were cultured from the second animal. This report confirms the pathogenicity and potentially fatal outcome of mycobacterial infection in manatees. In addition, the pathologic response to infection with these mycobacteria in manatees is similar to that associated with Mycobacterium spp. in other animals.     

Serum chemistry reference values in free‐ranging North Atlantic male walruses (Odobenus rosmarus rosmarus) from the Svalbard archipelago

Background: Information regarding health and disease is limited for walruses, a keystone species in arctic marine ecosystems. Serum chemistry analysis is a useful clinical tool for the health assessment of walruses, but only a few captive Pacific walruses have been evaluated. Objectives: The aim of this study was to determine serum chemistry reference values for free‐ranging male Atlantic walruses (Odobenus rosmarus rosmarus) on Svalbard and to assess potential differences in animals with low and high tissue levels of organic pollutants. Methods: Blood samples were collected from 17 wild, adult, male Atlantic walruses chemically immobilized with etorphine at eastern Svalbard (Norway). Serum was obtained for routine biochemical analysis as well as nonesterified free fatty acids (NEFA) and cortisol tests. Serum protein concentration was also measured by agarose gel electrophoresis. Results: Reference values (ranges) included alanine aminotransferase (12–51 U/L), aspartate aminotransferase (54–137 U/L), alkaline phosphatase (42–243 U/L), creatine kinase (32–506 U/L), lactate dehydrogenase (480–1322 U/L), amylase (0–23 U/L), lipase (68–298 U/L), total protein (68–91 g/L), albumin (25.3–34.8 g/L), creatinine (84–137 μmol/L), urea (8.2–19.9 mmol/L), bilirubin (0–4 μmol/L), cholesterol (4.4–7.3 mmol/L), NEFA (0.1–0.4 mmol/L), triglycerides (0.6–2.2 mmol/L), calcium (2.0–2.7 mmol/L), phosphorus (1.7–2.8 mmol/L), sodium (147–162 mmol/L), potassium (4.7–7.4 mmol/L), chloride (102–115 mmol/L), and cortisol (<28–214 nmol/L). Walruses exposed to high levels of organic pollutants (n=6) had significantly lower (P=.022) phosphorus concentration than those with low levels of pollutants (n=6). Conclusions: The clinical chemistry reference values determined in this study can serve as baseline data for future health‐related studies of walruses in a changing Arctic and may also be helpful for health evaluations of walruses in captivity. Impacts of the exposure of marine mammals to organic pollutants should be further investigated.     

Comparison of blood aminotransferase methods for assessment of myopathy and hepatopathy in Florida manatees (Trichechus manatus latirostris)

Muscle injury is common in Florida manatees (Trichechus manatus latirostris). Plasma aspartate aminotransferase (AST) is frequently used to assess muscular damage in capture myopathy and traumatic injury. Therefore, accurate measurement of AST and alanine aminotransferase (ALT) is important in managed, free-ranging animals, as well as in those rehabilitating from injury. Activities of these enzymes, however, are usually not increased in manatees with either acute or chronic muscle damage, despite marked increases in plasma creatine kinase activity. It is hypothesized that this absence of response is due to apoenzymes in the blood not detected by commonly used veterinary assays. Addition of coenzyme pyridoxal-5-phosphate (P5P or vitamin B6) should, therefore, result in higher measured enzyme activities. The objective of this study was to determine the most accurate, precise, and diagnostically useful method for aminotransferase measurement in manatees that can be used in veterinary practices and diagnostic laboratories. Additionally, appropriate collection and storage techniques were assessed. The use of an optimized commercial wet chemical assay with 100 micromol P5P resulted in a positive bias of measured enzyme activities in a healthy population of animals. However, AST and ALT were still much lower than that typically observed in domestic animals and should not be used alone in the assessment of capture myopathy and muscular trauma. Additionally, the dry chemistry analyzer, typically used in clinics, reported significantly higher and less precise AST and ALT activities with poor correlation to those measured with wet chemical methods found in diagnostic laboratories. Therefore, these results cannot be clinically compared. Overall, the optimized wet chemical method was the most precise and diagnostically useful measurement of aminotransferase in samples. Additionally, there was a statistically significant difference between paired serum and plasma measurement, indicating that separate reference intervals should be established for serum and plasma. Finally, storage of these enzymes at -70 degrees C for 1 mo resulted in up to a 25% decrease in enzymatic activity in manatee plasma.     

Hematocrit and plasma chemistry values in adult collared scops owls (Otus lettia) and crested serpent eagles (Spilornis cheela hoya)

In this study, we report hematocrit and plasma chemistry values for adult captive collared scops owls (Otus lettia) and crested serpent eagles (Spilornis cheela hoya). In particular, we address the gender-specific differences within these values. We measured hematocrit (HCT) and plasma chemistry values for uric acid (UA), plasma urea nitrogen (BUN), total protein (TP), albumin (ALB), glucose (GLU), cholesterol (CHO), triglyceride (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), total bilirubin (TBIL), creatine (CRE), creatine phosphokinase (CPK), amylase (AMY), calcium (CA), ionic phosphorous (IP) and sodium (NA), potassium (K) and chloride ions (CL) in 37 adult captive collared scops owls and 39 adult captive crested serpent eagles. Significant differences between the sexes were found for UA, GLU and CPK in the collared scope owls. UA and GLU concentrations were significantly higher (P<0.01 and P<0.05) among males than females, while the CPK concentration was significantly lower (P<0.05) in males. There were no significant differences in of all of the measured parameters between male and female eagles. These finding suggested that HCT and plasma chemistry values of raptors vary individually according to species and sex. Our results provide the 1st available reference data for ranges of plasma values in adult captive collared scops owls and crested serpent eagles, making them a potentially useful complementary diagnostic tool for veterinary care of individuals for both species in captivity.     

Weights, hematology, and serum chemistry of free-ranging brown boobies (Sula leucogaster) in Johnston Atoll, Central Pacific.

Hematologic and serum chemistry values are reported for 105 brown boobies (Sula leucogaster) from Johnston Atoll, Central Pacific. Hematocrit, estimated total plasma solids, total and differential white cell counts, serum glucose, calcium, phosphorus, uric acid, total protein, albumin, globulin, aspartate aminotransferase, and creatinine phosphokinase were analyzed. Hematologic and serum chemistry values varied with age and sex. Values were compared with those of red-footed boobies and other tropical and temperate marine pelecaniforms.     

Serological evidence of Toxoplasma gondii infection in captive marine mammals in Mexico

Toxoplasma gondii infection in marine mammals is important because they are considered as a sentinel for contamination of seas with T. gondii oocysts, and toxoplasmosis causes mortality in these animals, particularly sea otters. Serological evidence of T. gondii infection was determined in 75 captive marine mammals from four facilities in southern and central geographical regions in Mexico using the modified agglutination test (MAT). Antibodies (MAT, 1:25 or higher) to T. gondii were found in 55 (87.3%) of 63 Atlantic bottlenose dolphins (Tursiops truncatus truncatus), 3 of 3 Pacific bottlenose dolphins (Tursiops truncatus gillii), 2 of 4 California sea lions (Zalophus californianus), but not in 3 West Indian manatees (Trichechus manatus), and 2 Patagonian sea lions (Otaria flavescens). Seropositive marine mammals were found in all 4 (100%) facilities sampled. All marine mammals were healthy and there has not been any case of clinical toxoplasmosis in the facilities sampled for at least the last 15 years. The seroprevalence of T. gondii infection in marine mammals of the same species did not vary significantly with respect to sex and age. This is the first report on the detection of antibodies to T. gondii in marine mammals in Mexico.     

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%.     

Serum biochemical analytes in captive Amazonian manatees (Trichechus inunguis)

Background: Establishment of reference values for serum biochemical analytes is important for monitoring health and physiological status of captive animals. Objective: The purpose of this study was to measure and report ranges for serum biochemical analytes in Amazonian manatees (Trichechus inunguis). Methods: Blood samples were collected from 24 healthy captive Amazonian manatees that comprised a mixture of adults, subadults, and calves and males and females; serum analytes were measured and analyzed using a dry reagent bench‐top chemical analyzer. Comparisons were made between sexes and with previously published values of closely related species. Results: Medians and ranges (minimum–maximum) of values for the analytes were: lactate dehydrogenase (LDH), 151 (111–278) U/L (n=20); creatine kinase, 144 (76–315) U/L (n=11); alanine aminotransferase, 10 (2–28) U/L (n=18); aspartate aminotransferase, 14 (5–28) U/L (n=21); γ‐glutamyltransferase, 47 (36–73) U/L (n=21); amylase, 1428 (1010–1874) U/L (n=21); alkaline phosphatase, 73 (36–141) U/L (n=19); total protein, 6.8 (6.2–8.0) g/dL (n=24); albumin, 3.3 (2.6–4.1) g/dL (n=21); cholesterol, 188 (101–399) mg/dL (n=21); triglycerides, 126 (60–236) mg/dL (n=21); glucose, 47 (22–69) mg/dL (n=21); urea, 43 (21–69) mg/dL (n=21); uric acid, 1.1 (0.5–1.8) mg/dL (n=22); creatinine, 2.2 (1.5–3.3) mg/dL (n=22); total bilirubin, 0.2 (0.2–2.0) mg/dL (n=21); calcium, 12.7 (10.2–18.6) mg/dL (n=24); iron, 282 (207–457) μg/dL (n=13); and magnesium, 6.9 (4.3–8.9) mg/dL (n=20). With the exception of LDH, no differences were observed between sexes. Conclusions: The ranges obtained in this study provide important preliminary estimates for concentrations and activities of serum analytes in Amazonian manatees until a larger reference interval study can be conducted.     

Ultrastructure of the spermatozoa from a Florida manatee (Trichechus manatus latirostris)

Semen was opportunistically collected from a free-ranging, 10-year-old, 275 cm (total length) Florida manatee (Trichechus manatus latirostris) during rehabilitation treatments. Ultrastructure of the spermatozoa was examined by scanning and transmission electron microscopy and differed slightly from that described for other mammals. Comparisons to the manatee's closest phylogenetic relatives, the elephant and hyrax, were made. The manatee spermatozoa had a similar acrosome but a distinct annulus and lacked the dense bodies observed in the neck of the elephant spermatozoa. Additionally, manatee spermatozoa lacked the lateral vacuoles observed in the nuclear chromatin from of the hyrax spermatozoa. These data add to our understanding of manatees and allow for comparative studies with other species that may be useful in phylogenetic and reproductive studies.     

Hematology and serum chemistry of captive juvenile double-crested cormorants (Phalacrocorax auritus).

Hematologic and serum chemical values were obtained for double-crested cormorants (Phalacrocorax auritus) to improve clinical diagnosis of disease in this species. Blood samples were collected from 20 captive double-crested cormorants at 4 to 6 weeks of age. Hematocrit and leukocyte concentrations were determined in heparinized blood. Concentrations of sodium, potassium, chloride, bicarbonate, calcium, phosphorus, creatinine, glucose, uric acid, total protein, and albumin, and the activity levels of alkaline phosphatase, creatinine kinase, and aspartate aminotransferase were determined in serum. Total leukocyte concentrations in these double-crested cormorants were higher than the limited ranges reported for cormorants of other species, possibly due to subclinical infection with the liver trematode Amphimerus elongatus, and to differences in species and age.     

Comparative haematology and blood chemistry of endangered lizards (Gallotia species) in the Canary Islands

Blood samples were taken from the ventral coccygeal vein of 15 El Hierro giant lizards (Gallotia simonyi) (seven females and eight males), six La Gomera giant lizards (Gallotia bravoana) (four males and two females) and four Tenerife giant lizards (Gallotia intermedia) (two males and two females), and 31 blood parameters were measured. Among the haematological parameters there were significant differences between the three species in heterophils, azurophils and lymphocytes, but no significant differences in red blood cell count, white blood cell count, haemoglobin, packed-cell volume, monocytes, eosinophils and basophils. In terms of blood chemistry there were significant differences between the three species in cholesterol, triglycerides, glucose, sodium, chloride, urea, uric acid, total proteins, prealbumin, albumin and gamma globulins, but no significant differences in calcium, potassium, aspartate aminotransferase, alanine aminotransferase, creatine kinase, bile acids, alpha-1 and alpha-2 globulins and beta globulins.     

Comparison of serum and plasma for determination of blood biochemical values in Malaysian flying foxes (Pteropus vampyrus).

The effects of the anticoagulant sodium heparin and time of centrifugation on 20 biochemical analytes in the blood of Malaysian flying foxes (Pteropus vampyrus) were evaluated. Paired plasma and serum samples were centrifuged at 1 hr and 6 hr postcollection. Heparinization and time of centrifugation did not significantly affect albumin, cholesterol, triglycerides, amylase, blood urea nitrogen, creatinine, alanine aminotransferase, aspartate aminotransaminase, alkaline phosphatase, calcium, sodium, and total carbon dioxide levels. Plasma was associated with higher globulin and lower potassium values. Glucose and chloride levels decreased significantly over time, whereas phosphorus levels increased. Serum creatine kinase activity at 6 hr postcollection was significantly higher than the other creatine kinase means. Sodium levels were not significantly affected by sodium heparin as used as an anticoagulant in this study.     

Effect of age on reference intervals of serum biochemical values in kittens

OBJECTIVE: To determine the effect of age on reference intervals of serum biochemical values in kittens. DESIGN: Prospective clinical trial. ANIMALS: 55 kittens from 12 specific-pathogen-free queens. PROCEDURE: Kittens were allocated at birth into colostrum-fed (n = 27) and colostrum-deprived (28) groups. Blood was collected at birth and on days 1, 2, 4, 7, 14, 28, and 56. Serum samples were analyzed for activities of alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, creatine kinase, lactate dehydrogenase, gamma-glutamyltransferase, amylase, and lipase and for concentrations of albumin, total protein, bilirubin, urea nitrogen, creatinine, cholesterol, glucose, calcium, phosphorus, and triglycerides by use of an automated analyzer. Total serum solids concentrations were determined by use of refractometry. Serum IgG concentrations were quantified by use of radial immunodiffusion. RESULTS: For several analytes, reference intervals changed rapidly, most notably during the first few days of life. Reference intervals for alkaline phosphatase, creatine kinase, lactate dehydrogenase, and triglycerides were higher from birth to 8 weeks than adult reference intervals. Aspartate aminotransferase, bilirubin, urea nitrogen, and creatinine were higher than in adults at birth but were similar to or lower than adult reference intervals by 8 weeks. Compared with adult reference intervals, reference intervals for calcium and phosphorus concentrations were higher and for albumin and total protein concentrations were lower throughout the study period. CONCLUSIONS AND CLINICAL RELEVANCE: Important differences exist between reference intervals for serum biochemical values of neonatal and adult cats. Age-appropriate reference intervals should be used for accurate assessment of serum biochemical test results in cats.     

The elephants of Zoba Gash Barka, Eritrea: part 4. Cholelithiasis in a wild African elephant (Loxodonta africana).

A 4.0-kg cholelith was found within the abdominal cavity of a dead wild African elephant (Loxodonta africana) in Eritrea. Analysis of this cholelith by histochemistry, electron microscopy, electrospray mass spectroscopy, and energy-dispersive x-ray spectroscopy revealed it was composed of bile alcohols but no calcium, bilirubin, or cholesterol. Bacteria were also found in the cholelith. Similar, but smaller, bile stones have been identified previously in other wild African elephants and an excavated mammoth (Mammuthus columbi). Choleliths have been reported only once in a captive Asian elephant (Elephas maximus). Elephants, along with hyraxes (Procavia capensis) and manatees (Trichechus manatus), are unique among mammals in producing only bile alcohols and no bile acids, which may predispose them to cholelithiasis, particularly in association with bacterial infection. Dietary factors may also play an important role in cholelith formation.