The effect of fresh gas flow during induction of anaesthesia on sevoflurane usage: a quality improvement study.

Reducing fresh gas flow during inhalational anaesthesia results in cost savings and decreases environmental impact. We are interested in the influence of fresh gas flow on the early (induction) phase of overall fresh gas flow and vapour consumption. This stage is often excluded in studies of fresh gas flow. Data were collected from 3199 sevoflurane anaesthetics over an 11-month period in four operating theatres. We determined fresh gas flow at different stages of anaesthesia, and developed an explanatory model for the influence of the 'induction' period. Following a three-month collection of baseline data we emphasised the importance of the early phase to our department repeatedly over a two-week period. We explored the relationship between fresh gas flow and total vapour usage, and used a simple mathematical model to explore the effect of changes in the fresh gas flow and duration of the 'induction' phase. Mean fresh gas flow was 1.15 l.min-1 in the baseline period and 0.91 l.min-1 in the two months following our educational effort (p = 0.0005). In the following six months, mean fresh gas flow was 1.17 l.min-1 (p = 0.7726 compared with baseline). These results were driven by changes in both fresh gas flow and duration of the initial high-flow period. We found some correlation (R2  = 0.85) between overall fresh gas flow and vapour consumption; a 1 l.min-1 increase in fresh gas flow consumes an additional 18 ml.hr-1 of liquid sevoflurane. This preliminary study demonstrates that an episode of high fresh gas flow at the start of anaesthesia has a large and modifiable effect on overall fresh gas flow and vapour consumption. We also confirmed the linear relationship between fresh gas flow and vapour usage.

Effective size of a wild salmonid population is greatly reduced by hatchery supplementation.

Many declining and commercially important populations are supplemented with captive-born individuals that are intentionally released into the wild. These supplementation programs often create large numbers of offspring from relatively few breeding adults, which can have substantial population-level effects. We examined the genetic effects of supplementation on a wild population of steelhead (Oncorhynchus mykiss) from the Hood River, Oregon, by matching 12 run-years of hatchery steelhead back to their broodstock parents. We show that the effective number of breeders producing the hatchery fish (broodstock parents; N(b)) was quite small (harmonic mean N(b)=25 fish per brood-year vs 373 for wild fish), and was exacerbated by a high variance in broodstock reproductive success among individuals within years. The low N(b) caused hatchery fish to have decreased allelic richness, increased average relatedness, more loci in linkage disequilibrium and substantial levels of genetic drift in comparison with their wild-born counterparts. We also documented a substantial Ryman-Laikre effect whereby the additional hatchery fish doubled the total number of adult fish on the spawning grounds each year, but cut the effective population size of the total population (wild and hatchery fish combined) by nearly two-thirds. We further demonstrate that the Ryman-Laikre effect is most severe in this population when (1) >10% of fish allowed onto spawning grounds are from hatcheries and (2) the hatchery fish have high reproductive success in the wild. These results emphasize the trade-offs that arise when supplementation programs attempt to balance disparate goals (increasing production while maintaining genetic diversity and fitness).

Neoplasia and granulomas surrounding microchip transponders in Damaraland mole rats (Cryptomys damarensis).

Damaraland mole rats (Cryptomys damarensis) are among the longest-living rodents, with a maximum longevity of approximately 16 years. As one of the few mammals termed eusocial, these animals have been used in behavioral, genetic, metabolic, and physiologic research at the University of Connecticut since 1997. For individual identification at 3 to 4 months of age, mole rats were subcutaneously implanted with microchip transponders (11 mm in length) in the dorsal cervical region. In 2007, 2 of the 90 implanted adults, 10-year-old and 9-year-old females, developed subcutaneous masses at the site of the implant. Histopathological and immunohistochemical examinations revealed amelanotic melanoma and fibrosarcoma, respectively, with metastasis of the amelanotic melanoma. In 2008, a total of 3 adult males were castrated as part of a sex behavior study; 3 months later, all 3 castrated males developed subcutaneous masses around their implants, whereas none of the noncastrated males had masses. After an additional 9 months, these masses were found to be granulomas. To the authors' knowledge, this is the first report of neoplasia in this species. Both the tumors and the granulomas surrounded the microchip transponder.

Mortality of one-week-old chickens during naturally occurring Marek's disease virus infection.

Marek's disease (MD) is a disease of chickens that occurs worldwide and has serious economic consequences. MD can present as one of several forms, with the most commonly occurring forms being the lymphoproliferative diseases. Under experimental conditions, an early mortality syndrome has been recognized following infection by some but not all strains of MD virus (MDV). This is the first report of a confirmed case of mortality due to naturally occurring MDV infection in 1-week-old, nonvaccinated, chickens. Necrotizing lesions were observed in the bursa of Fabricius, lung, duodenum, jejunum, and proventriculus, and large intranuclear inclusion bodies were a striking feature in tissues with lesions in all birds. Immunohistochemical staining for the pp38 protein of MDV revealed abundant pp38 antigen in the affected tissues, confirming the presence of MDV within the lesions. PCR yielded an amplicon with 97% homology to the meq gene of MDV. No evidence of co-infection by either of the immunosuppressive agents chicken anemia virus and infectious bursal disease virus was detected.

The effect of predictive display on the control of step changes in effect site sevoflurane levels.

Graphical displays of past and future levels of drugs may be a useful adjunct to manual dosing. We have previously found that a display of predicted future values speeds step changes in end-tidal sevoflurane. In this study anaesthetists made step changes of 0.3% in effect site sevoflurane, with and without the display and as increases and decreases. We analysed 91 changes. When the predictive display was present, users made larger vaporiser dial changes of 3.9% vs 3.1% (95% CI for the difference -1.3% to -0.01%, p = 0.046) reflected in larger end-tidal changes (95% CI for the difference -0.009 vol% to -0.34 vol%, p = 0.06). There was no difference in the speed of change (220 vs 227 s (95% CI for the difference -51 to 32 s)), or in the accuracy of the change. In this study the predictive display influenced the magnitude of the step changes made by anaesthetists but did not affect the speed or overall accuracy of the change.

Isolation of West Nile virus from mosquitoes, crows, and a Cooper's hawk in Connecticut.

West Nile (WN) virus, a mosquito-transmitted virus native to Africa, Asia, and Europe, was isolated from two species of mosquitoes, Culex pipiens and Aedes vexans, and from brain tissues of 28 American crows, Corvus brachyrhynchos, and one Cooper's hawk, Accipiter cooperii, in Connecticut. A portion of the genome of virus isolates from four different hosts was sequenced and analyzed by comparative phylogenetic analysis. Our isolates from Connecticut were similar to one another and most closely related to two WN isolates from Romania (2.8 and 3.6 percent difference). If established in North America, WN virus will likely have severe effects on human health and on the health of populations of birds.

Rapid preclinical detection of sheeppox virus by a real-time PCR assay.

Sheeppox virus (SPPV) is a member of the Capripoxvirus (CaPV) genus of the Poxviridae family. Members of this genus, which also include goatpox and lumpy skin disease viruses, cause economically significant disease in sheep, goats, and cattle. A rapid diagnostic assay for CaPV would be useful for disease surveillance as well as for detection of CaPV in clinical samples and for outbreak management. Here we describe a fluorogenic probe hydrolysis (TaqMan) PCR assay designed for rapid detection of CaPV and tested on sheep experimentally infected with a virulent strain of SPPV. This assay can detect SPPV in buffy coats, nasal swabs, oral swabs, scabs, and skin lesions as well as in lung and lymph nodes collected at necropsy. This single-tube diagnostic assay can be performed in 2 h or less and can detect viral DNA in preclinical, clinical, and postmortem samples.

Fluorescent antibody test for rapid detection of West Nile virus antigen in avian tissues.

An indirect fluorescent antibody test (IFA) was developed to detect West Nile virus (WNV) antigens in tissues from avian species. The test samples used in the study consisted of 100 sets of tissues from dead crows that had been collected during the 2001 surveillance in Connecticut. The test tissues were punctured with a fine point Dacron cotton-tipped applicator and smeared in duplicate on 10-well diagnostic printed glass slides. Among several fixatives tested, 4% paraformaldehyde was the best. Reagent calibration for the IFA test was done in WNV-infected Vero cells and control uninfected Vero cells. Optimized antibody and fluorescent conjugate concentrations were then applied for the detection of WNV antigen on fixed tissue impression smears. Several tissues, including brain, heart, liver, kidney, and spleen were tested by the IFA test. The brain and heart seemed to be unsuitable for the test because of excessive background. Both virus isolation and reverse transcription-polymerase chain reaction (RT-PCR) were used for validation, with the latter technique having a higher sensitivity. Therefore, IFA results were compared with RT-PCR results. The diagnostic sensitivity was 96.8% for liver, 96.4% for kidney, and 100% for spleen. The diagnostic specificity was 69% for liver, 95.3% for kidney and 95.8 for spleen. The IFA test performed best with spleen and kidney. The IFA test described here is a useful, practical, and rapid test for screening for WNV.

Dual expression of p80 type I and p68 type II interleukin-I receptors on anterior pituitary cells synthesizing growth hormone.

Interleukin-1 alpha (IL-1 alpha) and IL-1 beta bind to either the p80 type I IL-1 receptor (IL-1RI) or the p68 type II IL-1R (IL-1RII) on both T and B lymphocytes. We and others have previously shown that the anterior pituitary gland also has specific high affinity binding sites for IL-1 alpha (Kd = approximately 1 nM) and expresses transcripts for both isoforms of the IL-1R. However, the identity of cells in the anterior pituitary gland that express the IL-1R and whether different populations of adenohypophyseal cells express different isoforms of the IL-1R remain unknown. Here we have used a combination of immunohistochemistry and histochemistry to localize IL-1RI and IL-1RII to specific target cells in the mouse anterior pituitary gland. Perfusion-fixed, paraffin-embedded sections of anterior pituitary gland were immunolabeled with well characterized monoclonal antibodies to either IL-1RI or IL-1RII and counterstained using a modified Gomori's method to document acidophils and basophils. Immunolabeling demonstrated that both IL-1RI and IL-1RII were abundantly expressed on a single population of anterior pituitary gland cells and that these cells could be classified on the basis of histochemical staining as a subpopulation of acidophils. The distribution, morphology, and proportion of cells immunolabeled for IL-1RI and IL-1RII were consistent with GH-synthesizing cells. To confirm this hypothesis, a modified indirect avidin-biotin complex, sequential peroxidase/alkaline phosphatase technique was used to label anterior pituitary gland cells with antibodies to IL-1RI followed by antibodies to IL-1RII, GH, PRL, or ACTH. The IL-1RI-positive cells predominately coexpressed IL-1RII and GH, but very little, if any, PRL or ACTH. These data establish that the predominant cell population in the murine anterior pituitary gland that constitutively expresses IL-1R stain as acidophils histochemically, is round to oval with dense granular cytoplasm and eccentric nuclei, synthesizes GH, and simultaneously expresses IL-1RI and IL-1RII isoforms.

The West Nile virus: its recent emergence in North America.

West Nile fever emerged in New York in the summer of 1999 when seven people, several horses and thousands of wild birds died. It was soon established that the human disease and the mortality of birds were related. Continued surveillance detected West Nile virus in mosquitoes, birds, horses, small mammals, bats and humans, and has shown its spread to several northeastern states. These events confirm the establishment of West Nile virus endemically in the United States.

Expression and localization of p80 and p68 interleukin-1 receptor proteins in the brain of adult mice.

The biological effects of interleukin-1 (IL-1) are mediated by two distinct receptors, the p80 type I IL-1 and p68 type II IL-1 receptor proteins (IL-1RI and IL-1RII, respectively), both of which have been recently co-localized to the growth hormone synthesizing cells of the adenohypophysis. Previous studies have shown that IL-1 can bind to specific structures in the central nervous system, but the distribution of IL-1RI and IL-1RII proteins in the adult mouse brain has not been reported. Here we have used immunohistochemistry to study the expression, distribution and cellular localization of both isoforms of the IL-1 receptor proteins in the adult mouse brain. Using a combination of processing techniques (AMeX fixation and cryosectioning), we have immunolabeled brain sections for each isoform of the IL-1R. Both isoforms are expressed in the CNS, particularly in neuronal soma of the granular layer of the dentate gyrus and pyramidal cells of fields CA1-CA4 of Ammon's horn of the hippocampus, in epithelial cells of the choroid plexus and ependymal layer, and in neuronal soma of Purkinje cells of the cerebellum. The IL-1RII isoform, but not IL-1RI, is expressed in specific neuronal soma and proximal cell processes of neurons of the paraventricular gray matter of the hypothalamus. These immunohistochemical data directly demonstrate the neuronal expression of both IL-1R proteins in situ. The distribution and cellular localization of IL-1R proteins in the CNS provide a molecular basis for understanding reciprocal interactions between the immune system and the brain.

Therapeutic vaccination with p24-VLP and zidovudine augments HIV-specific cytotoxic T lymphocyte activity in asymptomatic HIV-infected individuals.

This study evaluates the impact of therapeutic vaccination with p24-VLP and zidovudine on the induction or maintenance of HIV-specific cytotoxic lymphocyte activity in a cohort of asymptomatic patients with CD4 counts greater than 400 cells/microl. In a dummy, randomized, phase II clinical trial of the therapeutic vaccine, participants were randomized to one of three arms for 6 months: p24-VLP (500 microg) in alum monthly plus zidovudine 200 mg tds, alum adjuvant plus zidovudine, or p24-VLP plus placebo. Subjects were studied for a total of 52 weeks from baseline. Monitoring included viral load, CD4 and CD8 counts, markers of immune activation, delayed-type hypersensitivity (DTH) skin testing, and cytotoxic T lymphocyte (CTL) measurement. The nine subjects who received p24-VLP and zidovudine had an augmentation and/or broadening of their CTL response compared with baseline (p = 0.004). The eight subjects receiving p24-VLP and seven subjects receiving zidovudine did not have a statistically significant increase or broadening of CTL activity. The augmentation of the CTL response in the subjects who received p24-VLP and zidovudine was not associated with a decline in viral load or an increase in CD8 counts. This study suggests that HIV-specific CTL activity can be augmented in HIV-infected individuals receiving p24-VLP and zidovudine, supporting the hypothesis of therapeutic vaccination in the presence of antiretroviral therapy.

Feather folliculoma in a captive-bred barn owl (Tyto alba).

Proliferative cutaneous lesions are frequently encountered in examination of avian species. Tumors of the skin have been reported in many bird species, although comparatively less is known about the incidence of integumentary neoplasia in nondomesticated species than in production or companion species. An adult male, 9-yr-old, captive-bred barn owl (Tyto alba) was presented for examination of a proliferative skin mass of several months' duration on the left wing. An excisional biopsy was performed, and the owl recovered uneventfully. Light microscopic examination of tissue sections of the mass revealed a focal, unencapsulated, well-demarcated, multiloculated mass that consisted of variably sized cystic spaces lined by stratified squamous epithelium and containing concentric laminations of keratin and foci of feather differentiation. A histopathologic diagnosis of feather folliculoma was made. This neoplasm has not previously been described in owls. Its incidence and documentation are significant in that this neoplasm should be considered as a differential diagnosis of proliferative skin lesions in this raptorial species.

Clinical and immunohistochemical characterization of thymic lymphosarcoma in a heifer.

A 2-year-old Holstein heifer with a swollen brisket, jugular vein distention, muffled heart sounds, tachycardia, and free gas bloat was examined. Thymic lymphosarcoma was suspected based on a negative agar gel immunodiffusion test for bovine leukemia virus, presence of atypical lymphocytes in pleural fluid, and detection of a mass in the thoracic inlet. Right-sided cardiac catheterization was performed, and markedly increased jugular venous pressures (41 mm Hg) with a pressure gradient of 29 mm Hg immediately cranial to the heart indicated constriction of the cranial vena cava. Immunohistochemical staining of formalin fixed, paraffin-embedded tissue sections of the tumor using a rabbit antihuman T cell, CD3 polyclonal antibody confirmed that the neoplastic lymphocytes were of thymic origin.

Efficacy of ivermectin chewable tablets and two new ivermectin tablet formulations against Dirofilaria immitis larvae in dogs.

One hundred four heartworm-free Beagles less than 1 year old were studied to determine the efficacy of ivermectin chewable tablets and of 2 other ivermectin tablet formulations against heartworm larvae. At 30 days after SC inoculation of dogs with infective Dirofilaria immitis larvae, all ivermectin formulations were given orally at dosage of 6 micrograms/kg of body weight. The ivermectin chewable tablets also were given orally at dosage of 2 and 6 micrograms/kg at 30 and 45 days, respectively, after injection of larvae. Replicates of 6 or 8 dogs in each study were formed on the basis of gender and body weight and, within replicates, were randomly allocated to treatment groups. At 30 days after injection of larvae, the additional dogs (in replicates of 8) were assigned to the control group and to the group given ivermectin chewable tablets at dosage of 6 micrograms/kg. All dogs were housed individually. Necropsy was performed approximately 5 or 6 months after larvae were administered. In both trials, all control dogs had heartworms at necropsy (University of Illinois--geometric mean, 35.0; Florida--geometric mean, 26.1). In both trials, the ivermectin chewable tablet (6 micrograms/kg) and both tablet formulations (6 micrograms/kg) given at 30 days after larval injection, and the chewable formulation (6 micrograms/kg) given at 45 days after larval injection were 100% effective (P less than 0.01) in preventing development of induced infection with D immitis.(ABSTRACT TRUNCATED AT 250 WORDS)

Degenerative myeloencephalopathy in two llamas.

Degenerative myeloencephalopathy of unknown cause was diagnosed in 2 mature llamas. One llama had ataxia and paresis of the left limbs; whereas, the other developed tetraplegia that was not preceded by obvious ataxia or paresis. Results of cranial nerve examinations were unremarkable in both llamas. Cerebrospinal fluid from both llamas was normal, and hematologic and serum biochemical abnormalities were attributed to concurrent conditions and not to the myeloencephalopathy. Results of cervical spinal radiography and myelography in 1 llama were normal. In each llama, the diagnosis of degenerative myeloencephalopathy was made at necropsy after the llama failed to respond to treatment for other neurologic or muscular disorders. Lesions of degenerative myelopathy were found bilaterally in the white matter of all spinal cord segments, but were most pronounced in the thoracic segments. The dorsal and dorsolateral funiculi were most severely affected. The most prominent lesion was axonal sheath dilatation, with swelling or loss of axons. Dilated axon sheaths contained cellular and myelin debris, and accumulation of gitter cells was prominent. In the gray matter of 1 llama, a few degenerate neuronal cell bodies were seen in addition to axonal swelling. Degenerate neurons were identified in the brain stem of both llamas.

A ten-year audit of fresh gas flows in a New Zealand hospital: the influence of the introduction of automated agent delivery and comparisons with other hospitals.

Reducing fresh gas flow (FGF) rates with volatile anaesthetics reduces waste, with positive financial and environmental consequences. We have audited FGF since 2001 by analysis of data collected from anaesthetic machines. We recently introduced Aisys(®) (GE Healthcare, Madison, WI, USA) machines that allow automated control of end-tidal levels of volatile anaesthetics. In 2009 the mean FGF was 1.27 l/minute, which was lower than 2001 (2.05 l/minute) and 2006 (1.43 l/minute) and similar to two other New Zealand hospitals. Following introduction of the Aisys(®), mean FGF initially increased to 1.5 l/minute, but fell to 1.09 l/minute over the following 12 months. Median FGF showed a similar pattern and is now <600 ml/minute. Since introduction of the Aisys(®) workstation the proportion of time with flow rates >4 l/minute has increased. The proportion of time spent in automated delivery mode has increased from 35% to 63%. Users valued the workload reduction with end-tidal control. Our findings suggest that in daily practice, with a wide range of practitioners at different levels of training and a broad patient mix, mean flow rates of around 1.3 l/minute with median flows in the range 0.5 to 1.0 l/minute are achievable targets.

The development of a system to guide volatile anaesthetic administration.

We have developed and deployed within our operating rooms a system which provides real-time estimates of effect-site levels of inhalational anaesthetic agents along with forward predictions of end-tidal and effect-site concentrations. The initial aim of this project was to provide users of inhalational agents with tools similar to those available in target-controlled infusion systems. This paper describes the development and implementation of the system and outlines evaluation and uses of the system. The prototype was developed by combining a locally developed data logging and trend display system with a model of uptake developed as a teaching tool in 1982. This uptake model performs as well as contemporary models of propofol uptake and distribution. Following initial evaluation, the system has been deployed in over half our operating rooms and uses data gathered from the Datex/GE Anaesthesia Delivery Unit anaesthetic machines. We have conducted a number of studies of the system itself explored aspects of the underlying models, and used the system to investigate effect-site guided anesthesia and as a tool for data collection in other studies. The system has been well accepted locally and has been shown to facilitate faster changes in inhalational levels. We have also seen a significant decrease in fresh gas flow rates over recent years and attribute this in part to the predictive system, which simplifies the task of determining the appropriate combination of gas flow and vapour dial setting. The system also provides a useful platform for a range of research projects.

A New Zealand based cohort study of anaesthetic trainees' career outcomes compared with previously expressed intentions.

Predicting workforce requirements is a difficult but necessary part of health resource planning. A 'snapshot' workforce survey undertaken in 2002 examined issues that New Zealand anaesthesia trainees expected would influence their choice of future workplace. We have restudied the same cohort to see if that workforce survey was a good predictor of outcome. Seventy (51%) of 138 surveys were completed in 2009 compared with 100 (80%) of 138 in the 2002 survey. Eighty percent of the 2002 respondents planned consultant positions in New Zealand. We found 64% of respondents were working in New Zealand (P < 0.01). We found that family ties were an important influence on the choice of country of residence for 80% of New Zealand based respondents but only 40% of those living outside New Zealand agreed or strongly agreed with this statement (P < 0.01). Remuneration influenced country of residence for 76% of those living outside New Zealand but was important for only 2% of those resident in New Zealand (P < 0.01). Salaries in New Zealand were predominantly between NZ$150,000 and $200,000 while those overseas received between NZ$300,000 and $400,000. Of those that are resident in New Zealand, 84% had studied in a New Zealand medical school compared with 52% of those currently working overseas (P < 0.01). Our study shows that stated career intentions in a group do not predict the actual group outcomes. We suggest that 'snapshot' studies examining workforce intentions are of little value for workforce planning. However we believe an ongoing program matching career aspirations against career outcomes would be a useful tool in workforce planning.