Pharmacokinetics of intravenous propofol in southern white rhinoceros (Ceratotherium simum simum) after intramuscular etorphine-butorphanol-medetomidine-azaperone.

OBJECTIVE: To determine the pharmacokinetics of a single bolus of intravenous (IV) propofol after intramuscular administration of etorphine, butorphanol, medetomidine, and azaperone in 5 southern white rhinoceros to facilitate reproductive evaluations. A specific consideration was whether propofol would facilitate timely orotracheal intubation. ANIMALS: 5 adult, female, zoo-maintained southern white rhinoceros. PROCEDURES: Rhinoceros were administered etorphine (0.002 mg/kg), butorphanol (0.02 to 0.026 mg/kg), medetomidine (0.023 to 0.025 mg/kg), and azaperone (0.014 to 0.017 mg/kg) intramuscularly (IM) prior to an IV dose of propofol (0.5 mg/kg). Physiologic parameters (heart rate, blood pressure, respiratory rate, and capnography), timed parameters (eg, time to initial effects and intubation), and quality of induction and intubation were recorded following drug administration. Venous blood was collected for analysis of plasma propofol concentrations using liquid chromatography-tandem mass spectrometry at various time points after propofol administration. RESULTS: All animals were approachable following IM drug administration, and orotracheal intubation was achieved at 9.8 ± 2.0 minutes (mean ±SD) following propofol administration. The mean clearance for propofol was 14.2 ± 7.7 ml/min/kg, the mean terminal half-life was 82.4 ± 74.4 minutes, and the maximum concentration occurred at 2.8 ± 2.9 minutes. Two of 5 rhinoceros experienced apnea after propofol administration. Initial hypertension, which improved without intervention, was observed. CLINICAL RELEVANCE: This study provides pharmacokinetic data and insight into the effects of propofol in rhinoceros anesthetized using etorphine, butorphanol, medetomidine, and azaperone. While apnea was observed in 2 rhinoceros, propofol administration allowed for rapid control of the airway and facilitated oxygen administration and ventilatory support.

HYPERLIPIDEMIA AND XANTHOMATOSIS IN YELLOW-FOOTED ROCK WALLABIES (PETROGALE XANTHOPUS) UNDER MANAGED CARE.

Xanthomas are localized lipid deposits in organs with associated granulomatous inflammation. Xanthomatosis is a rare condition in both human and veterinary medicine and is often linked to inherited or acquired dyslipidemias. Three female yellow-footed rock wallabies (Petrogale xanthopus) at a single institution were diagnosed via biopsy with cutaneous xanthomas secondary to hypertriglyceridemia and hypercholesterolemia, and an additional two female yellow-footed rock wallabies were diagnosed with xanthomas at a second institution. All cases presented with cutaneous masses at the haired skin and paw pad junctions of the extremities, and/or mucocutaneous junctions of the face or urogenital tract. The clinically affected individuals were overconditioned or obese, had lipemic serum, and had elevations in blood cholesterol and triglyceride levels. When full lipid panels were performed, inverse high- and low-density lipoprotein fractions were observed. Six other individuals at the first institution had identical husbandry but were of more appropriate body condition, were normolipidemic, and had no xanthomas. One of the affected animals was also concurrently diagnosed with hepatic lipidosis via liver biopsy. Pedigree review and evaluation for underlying endocrine diseases such as hypothyroidism were performed. Because all affected animals were found to be related, a genetic predisposition is possible but requires further investigation. Consideration for the predisposition of some individuals for obesity, hyperlipidemia, and subsequent xanthoma formation should be factored in the husbandry and medical management of this species.

Eumycetoma and disseminated phaeohyphomycosis in a Sumatran tiger.

A 16-y-old female Sumatran tiger (Panthera tigris sumatrae) was evaluated for hyporexia. Examination revealed chronic kidney disease and a large subcutaneous axillary mass with draining tracts that contained numerous small black grains. Histologic examination revealed the presence of intralesional fungal hyphae. Persistent hyporexia and pyogranulomatous disease, as well as progressive cachexia and azotemia occurred despite treatment, and euthanasia was performed. Disseminated phaeohyphomycosis was diagnosed on postmortem examination, additionally affecting various lymph nodes, the nasal cavity, mesenteric adipose tissue, abdominal aorta, pericardium, and kidney. Fungal culture from a deep-tissue sample isolated a pure growth of Curvularia sp., a dematiaceous opportunistic fungus able to cause eumycetomas and/or phaeohyphomycosis. Phaeohyphomycosis is a rare but emerging condition, not previously reported as disseminated disease in an exotic carnivore, to our knowledge. Aggressive systemic antifungal treatment was unsuccessful, likely complicated by diagnostic challenges and concurrent renal disease. The presence of a swelling with abundant grains exiting draining tracts should direct clinicians to the diagnosis of a mycetoma, warranting early and aggressive treatment.

USE OF A HUMAN INDIRECT IMMUNOFLUORESCENCE ANTIBODY ASSAY FOR BALAMUTHIA MANDRILLARIS IN A GROUP OF CAPTIVE NORTHWEST BORNEAN ORANGUTANS (PONGO PYGMAEUS PYGMAEUS).

Granulomatous amoebic encephalitis caused by the free-living amoeba Balamuthia mandrillaris is a highly fatal disease that was first isolated from a mandrill (Mandrillus sphinx), and has since been diagnosed in several nonhuman primates including orangutans. Indirect immunofluorescence antibody (IFA) techniques for Balamuthia have been used in the fields of human medicine and epidemiology both for exposure assessment and screening of clinical patients for antemortem diagnosis. Stored serum samples from five captive Northwest Bornean orangutans (Pongo pygmaeus pygmaeus), including one who had died from B. mandrillaris infection, housed at a single facility were screened with a human IFA assay for B. mandrillaris. Only the single, clinically affected individual was seropositive, and the results suggest that the use of the available human B. mandrillaris IFA assay is a novel diagnostic option for detection of Balamuthia antibodies in this species. A validated screening serological test could be used in individuals exhibiting signs consistent with granulomatous amoebic encephalitis to facilitate earlier antemortem diagnosis of Balamuthia infection, which is critical if treatment is to be pursued. This pilot study presents the use of serological detection methods for B. mandrillaris screening in a nonhuman primate. Subsequent use of the B. mandrillaris IFA assay in the larger captive population should be pursued for validation of the test and to provide further information on seroprevalence and evaluation of risk factors for exposure to Balamuthia and subsequent development of disease.

COMPARISON OF FECAL CYTOLOGY AND PRESENCE OF CLOSTRIDIUM PERFRINGENS ENTEROTOXIN IN CAPTIVE BLACK-FOOTED FERRETS (MUSTELA NIGRIPES) BASED ON DIET AND FECAL QUALITY.

The black-footed ferret (Mustela nigripes) is an endangered mustelid native to North America. Gastroenteritis is a documented cause of morbidity and mortality in managed individuals, particularly by infectious agents. Fecal cytology is an inexpensive and rapid test that can help guide clinical management strategies for animals with enteritis; however, normal parameters have not been established in black-footed ferrets. The objective of this study was to characterize fecal cytological findings of 50 fecal samples from 18 black-footed ferrets that received two different diet types (ground meat versus whole prey) and that were visibly judged to be normal or abnormal. This study also tested for the presence of Clostridium perfringens enterotoxin by enzyme-linked immunosorbent assay in all abnormal and a subset of normal fecal samples. Significantly higher spore-forming bacteria and yeast prevalence were present in normal feces from individuals following the meat-based compared with the whole-prey diet. Samples from individuals with abnormal feces had significantly more spore-forming bacteria than normal feces, regardless of diet. Normal feces had higher diplococci and spore-forming bacteria compared with domestic canine and feline standards. A single abnormal fecal sample was positive for enterotoxin and originated from the only animal requiring treatment. Results indicate that low numbers of spore-forming bacteria can be found in fecal samples from clinically normal black-footed ferrets. Fecal cytology shows significantly increased spore-formers in clinically abnormal ferrets and in clinically normal ferrets following a ground meat-based diet.

Elucidating genomic regions determining enhanced leaf growth and delayed senescence in elevated CO2.

Limited information is available on the genetic variation and control for plant growth response to elevated CO(2) (e[CO(2)]). Such information is necessary to understand plant adaptation and evolution in future rising CO(2). Here, quantitative trait loci (QTL) for leaf growth, development, quality and leaf senescence were determined in a tree pedigree - an F(2) hybrid of Populus trichocarpa T. & G and Populus deltoides Marsh, following season-long exposure to either current day ambient carbon dioxide (a[CO(2)]) or e[CO(2)] at 600 microL L(-1). Leaf growth and development differed between the grandparents such that P. trichocarpa showed greater response to e[CO(2)]. In the F(2) generation, leaf development and quality traits including leaf area, leaf shape, epidermal cell area, and stomatal number, specific leaf area (SLA), and the phenology trait, canopy senescence index, were sensitive to e[CO(2)]. Sixty-nine QTL were mapped for the 19 traits of plants in a[CO(2)] while 60 QTL were mapped for plants in e[CO(2)]. The results suggest that although many QTL mapped to common positions in a[CO(2)] and e[CO(2)], confirming their importance in determining growth, there was also differential genetic control for a number of traits including leaf senescence. Candidate genes were shown to collocate to regions where response QTL mapped. This study is the first to identify candidate genes that may be important in determining plant adaptation to future high-CO(2) world.

Leaf stomatal and epidermal cell development: identification of putative quantitative trait loci in relation to elevated carbon dioxide concentration in poplar.

Genetic variation in stomatal initiation and density, and epidermal cell size and number were examined in a hybrid pedigree of Populus trichocarpa T. & G. and P. deltoides Marsh in both ambient ([aCO2]) and elevated ([eCO2]) concentrations of CO2. We aimed to link anatomical traits with the underlying genetic map of F2 Family 331, composed of 350 markers across 19 linkage groups. Leaf stomatal and epidermal cell traits showed pronounced differences between the original parents. We considered the following traits in the F2 population: stomatal density (SD), stomatal index (SI), epidermal cell area (ECA) and the number of epidermal cells per leaf (ECN). In [eCO2], adaxial SD and SI were reduced in the F2 population, whereas ECA increased and ECN remained unchanged. In [aCO2], four putative quantitative trait loci (QTL) with logarithm of the odds ratio (LOD) scores greater than 2.9 were found for stomatal traits on linkage group B: adaxial SI (LOD scores of 5.4 and 5.2); abaxial SI (LOD score of 3.3); and SD (LOD score of 3.2). These results imply that QTL for SI and SD share linkage group B and are under genetic control. More moderate LOD scores (LOD scores >/= 2.5) suggest QTL for SI on linkage groups A and B and for SD on linkage groups B, D and X with a probable co-locating quantitative trait locus for SI and SD on linkage group D (position 46.3 cM). The QTL in both [aCO2] and [eCO2] for adaxial SD were co-located on linkage group X (LOD scores of 3.5 and 2.6, respectively) indicating a similar response across both treatments. Putative QTL were located on linkage group A (position 89.2 cM) for both leaf size and ECN in [aCO2] and for ECA at almost the same position. The data provide preliminary evidence that leaf stomatal and cell traits are amenable to QTL analysis.

Contrasting effects of elevated CO2 and water deficit on two native herbs.

This study investigated the effects of carbon dioxide (CO)2 enrichment and soil water deficit on the water use efficiency (WUE) and growth of Sanguisorba minor Scop, (salad burnet I and Anthyllis vulneraria L. (kidney vetch), growing in controlled environments. Instantaneous WL E (IWUE) increased in both species in elevated CO2 , with a higher average increase in unwatered (UW) A. vulneraria over the drying cycle. Total plant WUE of A. vulneraria increased in elevated CO, and under water deficit: the UW plants in elevated CO., had higher WUE and reduced water loss. By contrast, thee was only an effect of water supply on S. minor: total plant WUE increased and water loss decreased in the UV plants in both CO2 , treatments. Total apparent root length (ARL) of both species increased with CO2 , enrichment and in UW S. minor total ARL was increased. By contrast, for A. vulneraria, total ARL of UV plants increased in ambient CO2 , but decreased in elevated CO2 as compared with well-watered (WW) plants. Shoot dry weight (SDW) and root dry weight increased in both species (WW and UW) with CO2 . enrichment. For UW S. minor, SDW decreased relative to WW plants in both CO2 treatments. By contrast, ANOVA showed no significant effect of water supply on SDW of A. vulneraria. Leafier length increased in both species in elevated CO2 , and decreased following drought. Cell wall tensiometric extensibility (%P) increased in expanding leaves of S. minor in elevated CO., and for both species %P decreased in the UW plants as compared with those WW. Leaf water potential (f) of both species was lower in growing leaves of WAV plants in elevated CO2 Water deficit reduced the Ψ of growing leaves in both CO2 , treatments. The different responses of these species suggest that in a drier, enriched CO2 , environment survival in a community might depend on their ability to maintain growth at the same time as conserving water.

Elevated CO2 , water relations and biophysics of leaf extension in four chalk grassland herbs.

Diurnal measurements of leaf or leaflet, extension, water relations and cell wall extensibility (ø) were made on young growing leaves of four chalk downland herbs (Sanguisorba minor Scop., Lotus corniculatus L., Anthyllis vulneraria L. and Plantago media L.) growing in controlled environment cabinets and exposed to either ambient or elevated CO2 . This study revealed differences in that effect of CO2 and the- control of leaf growth between the four species. Leaf extension rate (LER) increased significantly at night (average over 8 h) in elevated C02 for S. minor A. vulneraria and P. Media with a significant increase over the first 4h of darkness for S. minor, L, corniculatus and P. media, whilst for S. minor and P. media average day-time LER (over 16 h) also increased significantly in elevated CO2 , as compared with ambient CO2 . Water potential (ψpS), solute potential (ψpS), turgor pressure (P), yield turgor (Y) and the effective turgor for growth (Pe) were measured using psychrometers. Bollrte potentials of s, minor, A. vulneraria and P. media decreased significantly following exposure to elevated co2 with a Significant reduction in ψpS, during the day in A. Vulneraria. Turgor pressure increased significantly in elevated CO2 as compared with ambient CO2 in A. vulneraria but there was no effect of elevated CO2 on P in the uther species. effects of CO2 on ψpS; or ψpS; were observed, Leaf cell wall extensibility (ø) increased significantly in leaves of V. minor, L. corniculatus and P. exposed to elevated Co2 . whereas in A. vulneraria, there was no effect of CO2 on extensibility. These results suggest that the mechanism by which elevated CO2 promotes leaf growth differs between species arisce in S. minor, L. corniculatus, and p media. CO2 through an influence on cell wall properties, whist in A vulneraria higher values of P explain that increased leaf growth in elevated CO2 for this species.

Contrasting effects of elevated CO2 on the root and shoot growth of four native herbs commonly found in chalk grassland.

The aim of this study was to investigate the impact of ambient (345 µl l-1 ) and elevated (590 µl l-1 ) CO2 on the root and shoot growth of four native chalk grassland herbs: Sanguisorba minor Scop, (salad burnet), Lotus carniculatus L. (birdsfoot trefoil), Anthyllis vulneraria L. (kidney vetch) and Plantago media L. (hoary plantain). Elevated CO2 had contrasting effects on both shoot and root growth of the four species studied. Both leaf expansion and production were stimulated by elevated CO2 for S. minor, L. corniculatus and P. media, whilst for A. vulneraria, only leaflet shape appeared to be altered by elevated CO2 , with the production of broader leaflets, compared with those produced in ambient CO2 . After 100 d shoot biomass was enhanced in elevated CO2 for S. minor and L. corniculatus, whilst there was no effect of elevated CO2 on shoot biomass for A. vulneraria or P. media. Contrasting effects of CO2 were also apparent for measurements of specific leaf area (SLA), which increased for L. corniculatus, decreased for A. vulneraria and remained unaltered for S. minor and P. media in elevated compared with ambient CO2 . Elevated CO2 also had contrasting effects on both the growth and morphology of roots. The accumulation of root biomass was stimulated following exposure to elevated CO2 for S. minor and L. corniculatus whilst there was no effect on root biomass for A. vulneraria or P. media. Root length was measured on three occasions during the 100 d and revealed that exposure to elevated CO2 promoted root extension in S. minor, L. corniculatus and P. media, but not in A. vulneraria. Specific root length (SRL, length per unit dry weight) was increased in elevated CO2 for one species, P. media, whilst the root to shoot ratio of all four species remained unchanged by CO2 . These results show that four native herbs differ in their response to CO2 , suggesting that the structure of this plant community may be altered in the future.