We have developed a quadrupole magnetic flow sorter (QMS) to facilitate high-throughput binary cell separation. Optimized QMS operation requires the adjustment of three flow parameters based on the immunomagnetic characteristics of the target cell sample. To overcome the inefficiency of semiempirical operation/optimization of QMS flow parameters, a theoretical model of the QMS sorting process was developed. Application of this model requires measurement of the magnetophoretic mobility distribution of the cell sample by the cell tracking velocimetry (CTV) technique developed in our laboratory. In this work, the theoretical model was experimentally tested using breast carcinoma cells (HCC1954) overexpressing the HER-2/neu gene, and peripheral blood leukocytes (PBLs). The magnetophoretic mobility distribution of immunomagnetically labeled HCC1954 cells was measured using the CTV technique, and then theoretical predictions of sorting recoveries were calculated. Mean magnetophoretic mobilities of (1-3) x 10(-4) mm(3)/(T A s) were obtained depending on the labeling conditions. Labeled HCC1954 cells were mixed with unlabeled PBLs to form a "spiked" sample to be separated by the QMS. Fractional recoveries of cells for different flow parameters were examined and compared with theoretical predictions. Experimental results showed that the theoretical model accurately predicted fractional recoveries of HCC1954 cells. High-throughput (3.29 x 10(5) cells/s) separations with high recovery (0.89) of HCC1954 cells were achieved.

Infectious disease and other health hazards have been hypothesized to pose serious threats to the persistence of wild ape populations. Respiratory disease outbreaks have been shown to be of particular concern for several wild chimpanzee study sites, leading managers, and researchers to hypothesize that diseases originating from and/or spread by humans pose a substantial risk to the long-term survival of chimpanzee populations. The total chimpanzee population in Gombe National Park, Tanzania, has declined from 120-150 in the 1960s to about 100 by the end of 2007, with death associated with observable signs of disease as the leading cause of mortality. We used a historical data set collected from 1979 to 1987 to investigate the baseline rates of respiratory illness in chimpanzees at Gombe National Park, Tanzania, and to analyze the impact of human-related factors (e.g., banana feeding, visits to staff quarters) and non-human-related factors (e.g., sociality, season) on chimpanzee respiratory illness rates. We found that season and banana feeding were the most significant predictors of respiratory health clinical signs during this time period. We discuss these results in the context of management options for the reduction of disease risk and the importance of long-term observational data for conservation.

Despite the growing body of literature examining the welfare of zoo-housed animals, little standardized work has been published on the effect of construction and environmental disruption on the physiology and behavior of affected animals. When Lincoln Park Zoo (Chicago, IL), embarked on a renovation project for its Kovler Lion House, the opportunity was taken to perform a scientific study of behavioral and physiological markers in the resident felids to determine the effect of construction and environmental disruption. Fecal samples and behavioral observations were collected on four felid species (five individuals) before, during, and after the period of construction. As a group, the average z-score for fecal glucocorticoid metabolite concentration increased during construction relative to baseline. Levels remained elevated after construction, but trended toward baseline. All individuals demonstrated a significant decrease in the frequency of pacing and time spent visible during construction. Overall activity levels also showed a significant decrease relative to baseline measures. As zoological institutions continue to recognize the importance of habitat design, construction and renovation become inevitable. It is important to be aware of the potential consequences this can have on animals in the vicinity and to work toward minimizing negative effects. One recommendation is the availability of ample retreat and hiding space for felids during disruption to their environment.

Proper antibody labeling is a fundamental step in the positive selection/isolation of rare cancer cells using immunomagnetic cell separation technology. Using either a two-step or single-step labeling protocol, we examined a combination of six different antibodies specific for three different antigens (epithelial specific antigen, epithelial membrane antigen, and HER-2/Neu) on two different breast cancer cell lines (HCC1954 and MCF-7). When a two-step labeling protocol was used (i.e., anti-surface marker-fluoroscein-isothiocyanate [FITC] [primary Ab], anti-FITC magnetic colloid [secondary Ab]) saturation of the primary antibody was determined using fluorescence intensity measurements from flow cytometry (FCM). The saturation of the secondary antibody (or saturation of a single-step labeling) was determined using magnetophoretic mobility measurements from cell tracking velocimetry (CTV). When the maximum magnetophoretic mobility was the primary objective, our results demonstrate that the quantities necessary for antibody saturation with respect to fluorescence intensity were generally higher than those recommended by the manufacturer. The results demonstrate that magnetophoretic mobility varies depending on the types of cell lines, primary antibodies, and concentration of secondary magnetic colloid-conjugated antibody. It is concluded that saturation studies are a vital preparatory step in any separation method involving antibody labeling, especially those that require the specificity of rare cell detection.

Mycobacterium spp. infection is an important health concern for Asian elephant (Elephas maximus) populations worldwide. The disease is of particular concern considering its potential to affect not only the individual animal but also herd and public health. Although elephant tuberculosis susceptibility is poorly understood, immune function alterations are central to disease pathogenesis in other species and probably affect outcome of mycobacterial infections in elephants. Measurement of immune mediator (cytokine) levels within blood samples can provide information regarding immune function that may elucidate disease susceptibility. For this study, mRNA levels of interleukin (IL)-2, IL-4, IL-10, and IL-12; interferon (IFN)-gamma; tumor necrosis factor (TNF)-alpha; and transforming growth factor (TGF)-beta were measured using elephant-specific, real-time reverse transcription-polymerase chain reaction (RT-PCR) assays in RNA-preserved whole blood samples from 106 Asian elephants, 15% of which were Mycobacterium tuberculosis complex seropositive. The Elephant TB STAT-PAK (Chembio Diagnostics, Inc., Medford, New York 11763, USA), a novel lateral flow antibody detection assay developed for specific use in elephants, was used to determine serologic status for the study. Seropositive animals had higher levels of TNF-alpha and lower levels of TGF-beta than seronegative animals; these differences between groups were statistically significant when levels were analyzed as categorical variables. Trends toward higher levels of IFN-gamma and IL-4 and slightly lower levels of IL-10 and IL-12 were noted in the seropositive group, although differences between groups were not statistically significant. Presence of other inflammatory conditions was found to be a significant confounding variable in the analysis of the relationship between tuberculosis status and TNF-alpha levels, necessitating its inclusion in statistical models. Age and sex were not found to significantly affect the relationship between tuberculosis status and any of the cytokines measured. Interleukin-2 levels were below the sensitivity of the real-time RT-PCR assay irrespective of tuberculosis status. These findings provide a foundation for future research into the immunopathogenesis of elephant tuberculosis.