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.

Adrenocortical responsiveness in southern elephant seal mothers and pups during lactation and the effect of scientific handling.

We examined the cortisol responses to chemical and physical restraint stress in southern elephant seal Mirounga leonina females and their pups at three stages during lactation. In anaesthetised females the serum cortisol levels changed moderately during the 45-min sampling period following restraint, with average peaks at 23 min after anaesthetic administration. Overall, cortisol was relatively low 2 days postpartum and increased throughout lactation. In physically restrained pups serum cortisol increased rapidly after capture; the response was milder at age 2 days than at 11 days and 21 days. Levels were higher in female pups than in males. In order to test whether cortisol levels and/or responses became chronically (i.e. days to weeks) altered due to restraint, we compared the cortisol response at a late stage of lactation between three groups of mother-pup pairs previously given different levels of chemical (mothers) or physical (pups) restraint stress: control (not handled previously), moderate treatment (previously handled twice), and high treatment (previously handled 3-4 times). Pups of the three treatment groups showed similar adrenocortical responses suggesting no chronic effect of repeated physical restraint, despite the clear acute effects. Mothers of the control and moderate treatment groups showed similar cortisol responses; however, mothers of the high treatment group showed significantly attenuated responses. This indicated that elephant seals tolerated moderate degrees of handling disturbance; however, repeated (3-4) chemical immobilisations in lactating females may reduce their adrenocortical responsiveness for a period of days or weeks.

Pharmacokinetics of intravenously administered ketamine in southern elephant seals (Mirounga leonina).

Southern elephant seals (Mirounga leonina) are large, potentially dangerous animals which must be restrained before study or treatment. However, chemical restraint is unpredictable, possibly because of differences in body composition during fasting ashore, and circulatory adaptations to enable diving. The pharmacokinetics of ketamine (1.1 mg/kg i.v.) was studied in 15 southern elephant seals which had come ashore on Macquarie Island. The animals were first sedated with pethidine (5 mg/kg i.m.) to allow intravenous access. There was great variability in the calculated pharmacokinetic parameters, possibly due to circulatory changes associated with periods of apnoea which characterise this animal's response to anaesthetics. The median values were similar to those reported for other species: distribution t1/2 = 2.5 min (range 1-11 min); elimination t1/2 = 43 min (range 17-108 min); apparent volume of distribution, 1474 ml/kg (range 830-9301 ml/kg); clearance, 33 ml/min/kg (range 12-57 ml/min/kg). This was the first investigation of drug disposition in any seal species. and the first in any free-ranging wild animal. It is important to obtain pharmacological data in animals in whom drugs are used, but pre-catheterised captive animals may provide less variable data.

Molecular population genetics of the southern elephant seal Mirounga leonina.

Southern elephant seals breed on sub-Antarctic islands and have a circumpolar distribution. We assayed mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) variation in the three main populations in the south Atlantic, south Indian, and south Pacific oceans, and a smaller continental population in South America. Population structure of mtDNA was strong and not consistent with isolation by distance. The nDNA loci, although less informative, were consistent with the mtDNA results. Geographic structure appears to be dominated by historical processes, not contemporary gene flow. Uncorrected levels of nucleotide diversity for mtDNA control region I (2.86%) and nDNA (0.09%) were similar to those in humans and mice. Mutation rates for control region I (75 x 10(-9) substitutions per site per year) and nDNA (1.23 x 10(-9)) were similar to those in other mammals. Female effective population size and total effective population size were roughly equal at approximately 4 x 10(4), indicating a twofold greater rate of drift for mtDNA. Effective breeding sex ratio of four to five females per male was estimated from nucleotide diversity and mutation rates for mtDNA and nDNA, and was much less than behavioral observations would suggest. There was no evidence for selection at any of the assayed loci.

Use of the respiratory stimulant doxapram in southern elephant seals (Mirounga leonina).

The use of doxapram to stimulate breathing was examined in southern elephant seals chemically restrained with ketamine and xylazine. Animals which were breathing spontaneously received doxapram (approximately 0.5, 1, 2, or 4 mg/kg) or saline into the extradural intravertebral vein. Doxapram caused a dose-dependent increase in the depth and rate of respiration which began within one minute, peaked after two minutes and lasted for up to five minutes. A dose of 2 mg/kg appeared to be safe and effective for the stimulation of respiration, while 4 mg/kg caused arousal and shaking. Doxapram (2 mg/kg) was tested on 14 occasions in animals which had developed apnoea during chemical restraint. Doxapram had no effect when administered into the extradural intravertebral vein and appeared to be of more benefit when administered directly into the lungs via an endotracheal tube, but it was not effective in all cases. There was evidence to suggest that the endotracheal tube prevented some of the animals from breathing. The effect of intubation and endotracheal doxapram administration was therefore examined in 19 apnoeic and 31 spontaneously breathing seals. Intubation induced apnoea in animals at low levels of chemical restraint and endotracheal doxapram was unreliable for the stimulation of breathing.

Immobilisation of southern elephant seals and leopard seals with cyclohexamine anaesthetics and xylazine.

Ketamine and xylazine were given to 55 southern elephant seals (Mirounga leonina) for stomach lavaging, and to three leopard seals (Hydrurga leptonyx). The elephant seals showed prolonged apnoea and two of them died owing to aspiration of stomach contents. Two of the leopard seals died from unknown causes. Tiletamine and zolazepam were given to five elephant seals and one leopard seal. Two of the elephant seals and the leopard seal died from unknown causes. Xylazine alone was administered to 34 leopard seals. Sedation was poor at low dose rates (less than 1.7 mg/kg) but four of the seals given higher dose rates died owing to the aspiration of stomach contents.

Microbial community structure and biomass estimates of a methanogenic Antarctic Lake ecosystem as determined by phospholipid analyses.

Phospholipid analyses were performed on water column particulate and sediment samples from Ace Lake, a meromictic lake in the Vestfold Hills, Antarctica, to estimate the viable microbial biomass and community structure in the lake. In the water column, methanogenic bacterial phospholipids were present below 17 m in depth at concentrations which converted to a biomass of between 1 and 7×10(8) cells/liter. Methanogenic biomass in the sediment ranged from 17.7×10(9) cells/g dry weight of sediment at the surface to 0.1×10(9) cells/g dry weight at 2 m in depth. This relatively high methanogenic biomass implies that current microbial degradation of organic carbon in Ace Lake sediments may occur at extremely slow rates. Total microbial biomass increased from 4.4×10(8) cells/ liter at 2 m in depth to 19.4×10(8) cells/liter at 23 m, near the bottom of the water column. Total nonarchaebacterial biomass decreased from 4.2 ×10(9) cells/g dry weight in the surface sediment (1/4 the biomass of methanogens) to 0.06×10(8) cells/g dry weight at 2 m in depth in the sediment. Phospholipid fatty acid profiles showed that microeukaryotes were the major microbial group present in the oxylimnion of the lake, while bacteria dominated the lower, anoxic zone. Sulfate-reducing bacteria (SRB) comprised 25% of the microbial population at 23 m in depth in the water column particulates and were present in the surface sediment but to a lesser extent. Biomass estimates and community structure of the Ace Lake eco-system are discussed in relation to previously measured metabolic rates for this and other antarctic and temperate ecosystems. This is the first instance, to our knowledge, in which the viable biomass of methanogenic and SRB have been estimated for an antarctic microbial community.

Effect of storage conditions on nitrosated, acylated, and oxidized pyridine alkaloid derivatives in smokeless tobacco products.

Very large concentration increases in nitrite (34-fold), nitrosated pyridine alkaloids, and related 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) (14- to 33-fold) occurred in moist snuff during storage at 24 degrees C for 52 weeks, whereas, decreases in all parent and some acylated pyridine alkaloids were observed in the same material. Nitrite concentrations in dry snuff decreased up to 90% during storage; increased contents of nitrosated alkaloids and NNK of 30 to 80% were also observed. Storage effects on chewing tobacco included a 75% increase in nitrite and small increases of nitrosated alkaloids and NNK. Sums of parent alkaloids in moist snuff decreased 24 and 54% after storage for 24 weeks at 24 and 32 degrees C, respectively, while sums of alkaloid derivatives increased, up to 36-fold for nitrosated alkaloids and NNK, 92% for acylated, and 133% for oxidized components. Levels of N'-nitrosonornicotine, NNK, and N'-nitrosoanatabine after 52 weeks' storage at 24 degrees C were 547, 41, and 296% higher, respectively, in ambient air-exposed moist snuff than in the nonexposed counterpart. A mathematical model was evaluated and used to calibrate nonlinear gas chromatography alkali bead detector response to the individual components. This permitted the use of a single analysis for all required individual compounds over a wide concentration range.

Prolonged and multiple immobilizations of the southern elephant seal using ketamine hydrochloride-xylazine hydrochloride or ketamine hydrochloride-diazepam combinations.

Thirty seven southern elephant seals (Mirounga leonina) were singularly or repeatedly immobilized with combinations of ketamine hydrochloride (HCl) and xylazine HCl or ketamine HCl and diazepam. Atropine sulphate was included in the drug combinations. To permit experimental procedures the seals were immobilized for periods of 30-330 min. The mean induction dose of ketamine HCl was 8.71 +/- 0.25 mg/kg (mean +/- SE). The mean induction time was 16.02 +/- 2.62 min. For the elephant seals immobilized for periods in excess of 180 min, the mean dose of ketamine HCl used per hr was 3.31 +/- 0.13 mg/kg/hr and the mean dose of ketamine HCl used per hr postinduction was 1.31 +/- 0.15 mg/kg/hr. The mean dose of diazepam used was 0.09 +/- 0.01 mg/kg and the mean dose of xylazine HCl was 0.41 +/- 0.01 mg/kg. Elephant seals were weighed on 20 occasions (weight range: 897-1,932 kg) and the relationship between standard length and weight was found to be: Weight = 9.98 length - 2,317.63 (r2 = 0.724). Adverse reactions to seals immobilized only once or twice were not observed. Two seals immobilized on three occasions developed abscesses at the site of injection.

Effects of air-curing environment on alkaloid-derived nitrosamines in burley tobacco.

Levels of nitrite and pyridine alkaloid-derived total tobacco-specific nitrosamines (TSNA) were significantly higher in tobacco leaf (normal or late harvest) air-cured at 32 degrees C/83% relative humidity (RH) than in more moderate environments, i.e., 15 degrees C/50% RH and 24 degrees C/70% RH. These constituents increased appreciably from day 10 to day 21 of the cure. The near-concurrent appearances of maximal total contents of TSNA [sum of N'-nitrosonornicotine (NNN), N'-nitrosoanatabine (NAT), N'-nitrosoanabasine (NAB) and 4-(N-nitrosomethylamino)-1-(3-pyridyl)-1-butanone) (NNK)] and nitrite supports the concept that nitrite is a limiting and proximal precursor of total TSNA during the curing of tobacco. During a long curing period (50 days) at 32 degrees C/83% RH, nicotine and anatabine contents decreased, but TSNA contents increased in leaves of all harvest dates and stalk positions. These results support the view that nicotine and anatabine are precursors of TSNA. Measurement of nitrite and individual alkaloids during post-harvest processing of tobacco leaf may provide an index of the potential accumulation of alkaloid-derived nitrosamines.