Inferential and forward projection modeling to evaluate options for controlling invasive mammals on islands.

Successful pest-mammal eradications from remote islands have resulted in important biodiversity benefits. Near-shore islands can also serve as refuges for native biota but require ongoing effort to maintain low-pest or pest-free status. Three management options are available in the presence of reinvasion risk: (1) control-to-zero density, in which immigration may occur but reinvaders are removed; (2) sustained population suppression (to relatively low numbers); or (3) no action. Biodiversity benefits can result from options one and two. The management challenge is to make evidence-based decisions on the selection of an appropriate objective and to identify a financially feasible control strategy that has a high probability of success. This requires understanding the pest species population dynamics and how it will respond to a range of potential management strategies, each with an associated financial cost. We developed a two-stage modeling approach that consisted of (1) Bayesian inferential modeling to estimate parameters for a model of pest population dynamics and control, and (2) a forward projection model to simulate a range of plausible management scenarios and quantify the probability of obtaining zero density within four years. We applied the model to an ongoing, six-year trapping program to control stoats (Mustela erminea) on Resolution Island, New Zealand. Zero density has not yet been achieved. Results demonstrate that management objectives were impeded by a combination of a highly fecund population, insufficient trap attractiveness, and a substantial proportion of the population that did not enter traps. Immigration is known to occur because the founding population arrived on the island by swimming from the mainland. However, immigration rate during this study was indistinguishable from zero. The forward projection modeling showed that control-to-zero density was feasible but required greater than a two-fold budget increase to intensify the trapping rate relative to population growth. The two-stage modeling provides the foundation for a management program in which broad-scale trials of additional trapping effort or improved trap lures would test model predictions and increase our understanding of system dynamics.

A novel approach to assess the probability of disease eradication from a wild-animal reservoir host.

Surveying and declaring disease freedom in wildlife is difficult because information on population size and spatial distribution is often inadequate. We describe and demonstrate a novel spatial model of wildlife disease-surveillance data for predicting the probability of freedom of bovine tuberculosis (caused by Mycobacterium bovis) in New Zealand, in which the introduced brushtail possum (Trichosurus vulpecula) is the primary wildlife reservoir. Using parameters governing home-range size, probability of capture, probability of infection and spatial relative risks of infection we employed survey data on reservoir hosts and spillover sentinels to make inference on the probability of eradication. Our analysis revealed high sensitivity of model predictions to parameter values, which demonstrated important differences in the information contained in survey data of host-reservoir and spillover-sentinel species. The modelling can increase cost efficiency by reducing the likelihood of prematurely declaring success due to insufficient control, and avoiding unnecessary costs due to excessive control and monitoring.

High-throughput all-atom molecular dynamics simulations using distributed computing.

Although molecular dynamics simulation methods are useful in the modeling of macromolecular systems, they remain computationally expensive, with production work requiring costly high-performance computing (HPC) resources. We review recent innovations in accelerating molecular dynamics on graphics processing units (GPUs), and we describe GPUGRID, a volunteer computing project that uses the GPU resources of nondedicated desktop and workstation computers. In particular, we demonstrate the capability of simulating thousands of all-atom molecular trajectories generated at an average of 20 ns/day each (for systems of approximately 30 000-80 000 atoms). In conjunction with a potential of mean force (PMF) protocol for computing binding free energies, we demonstrate the use of GPUGRID in the computation of accurate binding affinities of the Src SH2 domain/pYEEI ligand complex by reconstructing the PMF over 373 umbrella sampling windows of 55 ns each (20.5 mus of total data). We obtain a standard free energy of binding of -8.7 +/- 0.4 kcal/mol within 0.7 kcal/mol from experimental results. This infrastructure will provide the basis for a robust system for high-throughput accurate binding affinity prediction.

Immigration policy, foreign agricultural labor, and exit intentions in the United States dairy industry.

United States agriculture is dependent on foreign labor. Current US immigration policies have been alleged to disrupt agricultural labor availability, particularly that of hired foreign labor. A national survey of dairy farmers across herd sizes and regions of the United States was conducted and the results were used to estimate the extent to which hired foreign labor dependency will affect exit intentions in dairy farming. This study found that the expected probability of exit from dairy farming increased as the use of hired foreign labor intensified. But the expected probability of exit also decreased rapidly as herd sizes got larger. Given the immigration policy currently in place, farmers expecting labor shortages in the future are expected to experience greater tendency to exit the industry.

Cost-based optimization of the stopping threshold for local disease surveillance during progressive eradication of tuberculosis from New Zealand wildlife.

Bovine tuberculosis (TB) is managed in New Zealand largely via population reduction of the major wildlife disease reservoir of Mycobacterium bovis, the introduced brushtail possum Trichosurus vulpecula. New Zealand aims to eradicate M. bovis infection from its livestock and wildlife within 40 years, as the culmination of progressive regional eradication programmes. Declarations of regional eradication are decided after extensive possum population control and post-control surveillance; hence, we developed a modelling framework, based on eco-epidemiological simulation data, to provide cost-evaluated options for deciding when to make these declarations. A decision-support framework evaluated potential costs of wildlife surveillance (and recontrol, if required) with respect to the calculated probability of successful eradication of M. bovis from wildlife. This enabled expected costs to be predicted in terms of stopping thresholds, allowing selection of optimal stopping rules based on minimizing costs. We identified factors that could influence optimal stopping values applied during regional eradication. Where vector/disease surveillance was inexpensive (for example, using low-cost detection devices or sentinel wildlife hosts) optimization involved setting a higher rather than lower stopping value, as it would be cheaper to minimize the risk of making a false declaration of eradication than to remedy any such failure. In addition, any cost of recontrol would largely depend on the time to rediscovery of residual M. bovis infection in wildlife, which would in turn be linked to the level of ongoing passive surveillance (with more rapid detection of re-emergent infection among wildlife in farmland situations than in remote forested regions). These two scenarios would favour different optimal stopping rules, as would the consideration of stakeholder confidence and socio-political issues, which are discussed. The framework presented here provides guidance to assess the economics underlying eradication of bovine TB from New Zealand farming; this eliminates reliance upon a pre-determined and uniform stopping rule for ceasing active management.

Livestock as sentinels for an infectious disease in a sympatric or adjacent-living wildlife reservoir host.

A central question to address in managing wildlife diseases is how much effort and resources are required to reduce infection prevalence to below a requisite threshold? This requires surveillance for infection in at least one species involved in the infection cycle, a process that is often expensive and time-consuming but one which could be enhanced using additional sources of readily-obtainable surveillance data. We demonstrate how surveillance data from ruminant livestock monitored for bovine tuberculosis (bTB) in New Zealand can be employed in spatially-explicit modelling to help predict the probability of freedom from Mycobacterium bovis infection in a sympatric wildlife reservoir species, the brushtail possum (Trichosurus vulpecula). We apply the model to a case study and compare resulting probabilities of freedom when utilizing (1) livestock data only, (2) wildlife data only, and (3) combined livestock-plus-wildlife surveillance data. Results indicated that the greatest probability of M. bovis eradication was achieved using wildlife monitoring data supplemented with livestock surveillance data. This combined approach lessened the time required for a confident (95% probability) declaration of regional eradication. However, the combined model was sensitive to the precision of the input parameters, and we describe ways to account for this. In a broad sense, this modelling approach is flexible in that any spatial arrangement of wildlife habitat and farms can be analysed, provided infection is readily detectable in both the wild and domestic animal(s) of interest. It is applicable to monitoring any communicable wildlife disease that affects regularly-tested livestock. The potential benefits to wildlife disease management include reduced surveillance costs and more rapid achievement of targeted reductions in disease prevalence.

The influence of taste in willingness-to-pay valuations of sirloin steaks from postextraction algal residue-fed cattle.

Consumer preferences and willingness-to-pay (WTP) for beef sirloin steaks with differing production, physical, and credence attributes related to the use of postextraction algal residue (PEAR), a novel feed ingredient, were estimated. Ninety-six consumers participated in a sensory tasting panel before completing a choice set survey; 127 consumers completed only the choice set survey without sampling products. Steaks from grain- and PEAR-fed steers had similar Warner-Bratzler shear force (WBSF) scores (1.89 kg and 2.01 kg, respectively; = 0.77) and had lower WBSF scores than steaks from grass-fed steers (3.37 kg; < 0.05). Eicosapentaenoic acid (20:5) was not different among steaks from grain- and PEAR-fed steers ( = 0.39) but was greater compared with steaks from grass-fed cattle ( ≤ 0.03). Panelists in the sensory portion of the study evaluated beef samples for like/dislike of overall sample, overall flavor, beefy flavor, and juiciness. Panelist rating of overall like, overall flavor like, and beefy flavor like were not different between the PEAR- and grain-fed treatments ( > 0.26). Panelists rated the juiciness like/dislike of steaks from PEAR-fed cattle the highest ( < 0.01) among the 3 samples. Sensory tasting of the products was observed to alter the preferences of consumers. Consumers who completed only the survey negatively perceived beef from PEAR-fed cattle compared with beef from grain-fed cattle, with a WTP discount of -US$1.17/kg. However, with sensory tasting, the WTP for beef from PEAR-fed cattle was not discounted relative to beef from grain-fed cattle ( = 0.21). The nontasting consumers had much higher stated WTP values for credence attributes. Factors that influence the eating experience (tenderness and quality grade) dominated as the most important attributes on WTP among the tasting group. The use of no hormones and no antibiotics in production had a premium of $2.34/kg among the nontasting group, but with tasting, the premium was $1.19/kg. If PEAR-fed beef came to market, there would be no need to differentiate it from grain-fed beef unless retailers wanted to market it as a differentiated product. If it were marketed as a differentiated product, retailers would need to hold promotional tastings to change consumer's preconceived notions about the product.

Development of integrated surveillance systems for the management of tuberculosis in New Zealand wildlife.

Disease surveillance for the management of bovine tuberculosis (TB) in New Zealand has focussed, to a large extent, on the development of tools specific for monitoring Mycobacterium bovis infection in wildlife. Diagnostic techniques have been modified progressively over 30 years of surveillance of TB in wildlife, from initial characterisation of gross TB lesions in a variety of wildlife, through development of sensitive culture techniques to identify viable mycobacteria, to molecular identification of individual M. bovis strains. Of key importance in disease surveillance has been the elucidation of the roles that different wildlife species play in the transmission of infection, specifically defining brushtail possums (Trichosurus vulpecula) as true maintenance hosts compared to those that are predominantly spillover hosts, but which may serve as useful sentinel species to indicate TB persistence. Epidemiological modelling has played a major role in TB surveillance, initially providing the theoretical support for large-scale possum population control and setting targets at which control effort should be deployed to ensure disease eradication. As TB prevalence in livestock and wildlife declined throughout the 2000s, more varied field tools were developed to gather surveillance data from the diminishing possum populations, and to provide information on changing TB prevalence. Accordingly, ever more precise (but disparate) surveillance information began to be integrated into multi-faceted decision-assist models to support TB management decisions, particularly to provide informed parameters at which control effort could be halted, culminating in the Proof of Freedom modelling framework that now allows an area to be declared TB-free within chosen confidence limits. As New Zealand moves from large-scale TB control to regional eradication of disease in the coming years, further integrative models will need to be developed to support management decisions, based on combined field data of possum and TB prevalence, sentinel information, risk assessment in relation to financial benefits, and changing political and environmental needs.

Vitamin A concentrations in liver determined by isotope dilution assay with tetradeuterated vitamin A and by biopsy in generally healthy adult humans.

The vitamin A status in 11 generally healthy surgical patients was estimated by measuring the dilution of a 45-mg oral dose of tetradeuterated retinyl acetate (99% pure). After purification of retinol by high-performance liquid chromatography, the ratio of 2H4-retinol:1H-retinol in plasma was measured by gas chromatography-mass spectrometry. On the basis of the observed ratios of [2H4]retinol:[1H]retinol over 19-47 d, the total body reserves and liver concentrations of vitamin A were calculated. Liver biopsy samples taken at surgery were directly analyzed for vitamin A. The correlation coefficient between calculated and measured liver vitamin A concentrations for 10 of the subjects was 0.88, and the Spearman's rank correlation coefficient was 0.95 (p less than 0.002). Thus, total body reserves of vitamin A in humans can be estimated validly in the marginal and satisfactory ranges by a benign, relatively noninvasive procedure.

Misdiagnosis of spasm of the near reflex.

A bilateral sixth nerve palsy portends serious disease of the central nervous system and precipitates extensive patient studies. Spasm of the near reflex, characterized by intermittent convergence, accommodation, and miosis, is a functional disturbance. Five patients with hysterical spasm of the near reflex erroneously diagnosed as a bilateral sixth nerve palsy are reported. The pupillary sign, intense miosis on attempted lateral gaze, is emphasized as an important clue to the correct diagnosis. Despite extensive investigation, no disease of the central nervous system was found. Neurotic or hysterical features were evident in every patient.

In vitro immunostimulation of rainbow trout (Oncorhynchus mykiss) spleen cells with levamisole.

An in vitro immunization and cultivation method for fish spleen organ sections was used to investigate the effects of levamisole on the immune response. After 10 days of culture with either 50 micrograms/ml, 25 micrograms/ml, 5 micrograms/ml or no levamisole in the media, the nonspecific defense reactions were measured by determining the metabolic activity of neutrophils by using the nitroblue tetrazolium test, and phagocytic and adherence indexes by incubating the fish cells with suspensions of formalin-killed Staphylococcus aureus. The specific immune system activity was shown by the passive hemolytic plaque assay demonstrating the numbers of antibody-producing cells. Elevations were found in nonspecific defense and specific immune response in the 5 micrograms/ml levels of levamisole. The 25 micrograms/ml levels instigated a slight elevation in some nonspecific levels but showed suppression in the specific immune response. The 50 micrograms/ml levels suppressed all indicators whether levamisole was given alone or combined with Yersinia ruckeri O-antigen or DNP-Ficoll. These antigens are themselves nonspecific defense stimulators, and a synergistic effect was evident when combined with 25 micrograms/ml and 5 micrograms/ml levels of levamisole.

Flush exposure and injection immunization of rainbow trout to selected DNP conjugates.

Rainbow trout (Salmo gairdneri) were immunized by flush exposure or intraperitoneal injection with single doses of the following dinitrophenyl (DNP) conjugates: DNP-keyhole limpet hemocyanin (KLH), DNP-Ficoll, DNP-O-antigen (from the fish pathogen, Yersinia ruckeri), DNP-sheep red blood cells, or DNP-duck red blood cells. The immune response was demonstrated by the passive hemolytic plaque assay to show splenic antibody-producing cells (APC) 14 days after antigen administration. High numbers of splenic APC specific for the hapten were found when the conjugates DNP-KLH, DNP-Ficoll, and DNP-O-antigen were given by injection and when DNP-O-antigen was given by flush exposure. The heterologous red blood cells were relatively nonimmunogenic. When the immune response to the hapten and carrier could be measured--e.g., after immunization with the DNP-O-antigen--the numbers of APC were consistently higher for the hapten.

Molecular analysis of the lymphocyte membrane.

Methods for the molecular analysis of lymphocyte membranes are reviewed briefly, and wherever possible presented in a manner relevant to comparative studies. The specific areas reviewed include the bulk isolation of lymphocyte membranes, the use of radioisotopes to covalently label lymphocyte membrane molecules, the use of lectins to characterize membrane glycoconjugates, and our current understanding of lymphocyte membrane immunoglobulins.

The immune response in immunized and naturally infected rainbow trout (Salmo gairdneri) to Diplostomum spathaceum as detected by enzyme-linked immunosorbent assay (ELISA).

The enzyme-linked immunosorbent assay (ELISA) procedure was modified and adapted for detection of circulating antibodies in rainbow trout (Salmo gairdneri) against metacercariae of the digenean trematode Diplostomum spathaceum, the causative agent for diplostomiasis. Rainbow trout (Salmo gairdneri) were injected with sonicated metacercariae representing 10, 40, and 100 metacercariae per fish. Three weeks after immunization the average titers for trout injected with 10, 40, and 100 metacercariae were 874, 841, and 525, and by six weeks the titers had fallen to 299, 349, and 203, respectively. Nine weeks after initial immunization, two remaining fish initially immunized with 100 metacercariae per fish were injected with a booster of 50 sonicated metacercariae per fish. Four weeks later the average titer was 1204. Serum samples from naturally infected wild fish tested for the presence of circulating antibodies against Diplostomum spathaceum showed 25 of 27 with positive titers.

In vitro antigen trapping by gill cells of the rainbow trout: an immunohistochemical study.

An in vitro assay was used to study the involvement of gill cells in the trapping and processing of particulate antigens. Gills were routinely processed for light microscopy after being placed in medium containing either Yersinia ruckeri O-antigen-labelled fluorescent beads, unlabelled fluorescent beads, Y, ruckeri O-antigen or formalin-killed Y. ruckeri, for 0, 30 s, 1, 5 and 30 min. Y. ruckeri formalin-killed cells, Y. ruckeri O-antigen and fluorescent beads labelled with Y. ruckeri O-antigen were taken in by gill epithelial cells as soon as 30 s after administration. In contrast, unlabelled fluorescent beads adhered to the epithelial cell membranes, but did not occur inside the gill cells. These results are discussed principally in relationship with the specificity of antigen trapping.

In vitro effect of levamisole on the neutrophil activity in rainbow trout (Oncorhynchus mykiss).

An in vitro immunization and cultivation method for fish spleen organ section was used to investigate the effects of levamisole on the neutrophil activity. After 10 days of culture with either 50 micrograms/ml, 25 micrograms/ml, 5 micrograms/ml or no levamisole in the media, the nonspecific defense reactions were measured by determining the metabolic activity of neutrophils by using the nitroblue tetrazolium test, and phagocytic and adherence indexes by incubating the fish cells with suspensions of formalin-killed Staphylococcus aureus. Elevations were found in nonspecific cellular defense in the 5 micrograms/ml levels of levamisole. The 25 micrograms/ml levels instigated a slight elevation in nonspecific cellular defense and the 50 micrograms/ml levels suppressed all indicators, whether levamisole was given alone or combined with Yersinia ruckeri O-antigen or DNP-Ficoll. These antigens are themselves nonspecific cellular defense stimulators, and a synergistic effect was evident when combined with 25 micrograms/ml and 5 micrograms/ml levels of levamisole.

Immunostimulants added to injected Aeromonas salmonicida bacterin enhance the defense mechanisms and protection in rainbow trout (Oncorhynchus mykiss).

Immunostimulants were given to rainbow trout for assaying effects on modulating non-specific defense mechanisms, specific immune response, and protection levels against pathogen challenge with Aeromonas salmonicida. Three drugs, levamisole (an approved veterinary drug in the USA), a quaternary ammonium compound (QAC), and a short-chain polypeptide (ISK) were found to affect the non-specific defense mechanism activities, which were measured by changes in circulatory neutrophil and phagocytic activity levels, and the specific immune response factors, which were measured by numbers of plaque-forming cells, and circulatory antibody levels. When given alone, the immunostimulants elevated the non-specific factors. When injected in combination with an A. salmonicida O-antigen bacterin, the non-specific factors were further elevated, and the specific response was raised over samples taken from fish given the bacterin without the immunostimulants. Challenge tests with the virulent pathogen, A. salmonicida, showed a 5-6 day delay in the onset of mortalities in the fish given the immunostimulants alone, and a 12-14 day delay when immunostimulants given were combined with the bacterin. In the groups given the QAC or ISK with the bacterin, there was a 20% and 40% survival rate, respectively.

Neutrophil, glass-adherent, nitroblue tetrazolium assay gives early indication of immunization effectiveness in rainbow trout.

Neutrophil activity in rainbow trout (Oncorhynchus mykiss) is increased upon antigenic stimulation with the Yersinia ruckeri O-antigen bacterin. The characteristics of neutrophil attachment to glass and nitroblue tetrazolium (NBT) staining were used to determine the effectiveness of immunization programs with fingerling rainbow trout. Fish immunized by intraperitoneal injection with doses of 100, 10, or 1 microgram of the bacterin showed the highest responses in that order in numbers of glass adherent, NBT-positive neutrophils. Studies on the kinetics of the occurrence of numbers of glass-adherent, NBT-positive staining cells from the fish injected with the 10 micrograms dose showed the numbers of positive cells were largest on Day 2 after injection. The specific immune response was confirmed by demonstrating the presence of plaque-forming cells by the passive hemolytic plaque assay and the rise in humoral antibody titers by passive hemagglutination 12 days after injection. The effects of immunization in trout could be detected earlier by using the neutrophil glass adherence and NBT reduction assays than by using assays based on observations of the specific immune response.

Dietary intake of immunostimulants by rainbow trout affects non-specific immunity and protection against furunculosis.

Immunostimulant preparations Macrogard, Candida utilis, Saccharomyces cerevisiae, Evetsel, Chitosan, or FinnStim were mixed into semipurified diets and fed to groups of rainbow trout for 1 week. Fish were bled by non-lethal caudal puncture and blood samples assayed for changes in non-specific cellular immunity and humoral protein levels. In the immunostimulated fish, hematocrit levels and lymphocyte counts remained relatively stable; however, elevations were observed in oxidative radical release, myeloperoxidase activity, phagocytic indexes, and potential killing activities of phagocytic cells including neutrophils. Total plasma protein and total immunoglobulin levels were elevated by the dietary immunostimulants. A challenge with the virulent pathogen that causes furunculosis, Aeromonas salmonicida, showed that the immunostimulated groups of fish were more resistant to the disease, confirming the potential use of these substances in fish culture for the prevention of disease.

Immunization and culture of rainbow trout organ sections in vitro.

Splenic and anterior kidney sections or whole organs were excised from large (1 kg) or small (200 g) rainbow trout (Salmo gairdneri) and placed in sterile 60 mm plastic plates containing 10 ml of Eagle's minimal essential medium (EMEM) supplemented with normal or fetal calf serum for in vitro culture. The organ samples were immunized in vitro by direct injection or by mixing in the medium Yersinia ruckeri O-antigen or dinitrophenyl-Ficoll. The medium was changed once during the 10-day incubation at 15 C. The passive hemolytic plaque assay demonstrated antibody production from the plaque-forming cells (PFC); passive hemagglutination was used to measure antibody titers in the media. High numbers of PFC occurred in cultures of either kidney or spleen, demonstrating that these organs can function independently for antibody production. Splenic sections from large fish produced more PFC than comparable whole organs from small fish. EMEM supplemented with 2% normal calf serum was a satisfactory culture medium. 2-hydroxyethyl-mercaptan an ingredient used in mammalian cell culture, inhibited antibody production in trout cells. These techniques are being used in the culture of organs and cells to elucidate pathways and sequences of antigen uptake and delivery of the immunopoietic tissues in trout.