Estimation of Species Identification Error: Implications for Raptor Migration Counts and Trend Estimation

Abstract: One of the primary assumptions associated with many wildlife and population trend studies is that target species are correctly identified. This assumption may not always be valid, particularly for species similar in appearance to co-occurring species. We examined size overlap and identification error rates among Cooper's (Accipiter cooperii) and sharp-shinned (A. striatus) hawks specific to a raptor migration count station along the Pacific Coast of North America. Illustrating the difficulty of distinguishing between these 2 species, we found overlap in 7 metrics among species-sex groups and in 2 metrics between species, and a principal components analysis revealed a continuum of discrete clusters for each species-sex combination in morphospace. Among juvenile hawks (n = 940), we found the greatest misidentification rate for male Cooper's hawks (23% of the 156 males were identified as sharp-shinned), lesser error rates for female Cooper's (8%, n = 339) and female sharp-shinned (6%, n = 246), and the lowest misidentification rate for male sharp-shinned hawks (0%, n = 199). We observed a similar pattern of misidentification among adult hawks (n = 48). We attempted to use conditional probabilities (identification rates) from calibration data to calculate the true number of adult and juvenile Cooper's hawks and sharp-shinned hawks. Discrepancies between total number of observed accipiters and estimated number using calibration data suggest that daily observer misclassification rates are higher than misclassification rates estimated from calibration data and prevent correction of the raw data. Our results illustrate the importance of testing for and quantifying observer error in species identification in wildlife census and population trend studies particularly when target species may be easily confused with other nontarget species.     

Constitutive Reactive Oxygen Species Generation from Autophagosome/Lysosome in Neuronal Oxidative Toxicity

Reactive oxygen species (ROS) are involved in several cell death processes, including cerebral ischemic injury. We found that glutamate-induced ROS accumulation and the associated cell death in mouse hippocampal cell lines were delayed by pharmacological inhibition of autophagy or lysosomal activity. Glutamate, however, did not stimulate autophagy, which was assessed by a protein marker, LC3, and neither changes in organization of mitochondria nor lysosomal membrane permeabilization were observed. Fluorescent analyses by a redox probe PF-H₂TMRos revealed that autophagosomes and/or lysosomes are the major sites for basal ROS generation in addition to mitochondria. Treatments with inhibitors for autophagy and lysosomes decreased their basal ROS production and caused a burst of mitochondrial ROS to be delayed. On the other hand, attenuation of mitochondrial activity by serum depletion or by high cell density culture resulted in the loss of both constitutive ROS production and an ROS burst in mitochondria. Thus, constitutive ROS production within mitochondria and lysosomes enables cells to be susceptible to glutamate-induced oxidative cytotoxicity. Likewise, inhibitors for autophagy and lysosomes reduced neural cell death in an ischemia model in rats. We suggest that cell injury during periods of ischemia is regulated by ROS-generating activity in autophagosomes and/or lysosomes as well as in mitochondria.     

Plant species effect on the spatial patterns of soil properties in the Mu-us desert ecosystem,Inner Mongolia,China

Although Artemisia ordosica Krasch. and Sabina vulgaris Ant. are the dominant shrub species in the Mu-us desert ecosystem, they differ in their botanical traits. We investigated the spatial patterns of soil properties using geostatistical analysis to examine the effect of plant species on these spatial patterns. Comparison among three microsite types (under A. ordosica, under S. vulgaris, and the opening between vegetation) showed that A. ordosica generally had less effect than S. vulgaris on local soil properties. The long life-span, prostrate life-form, and evergreen leaf-habit of S. vulgaris may lead to a higher accumulation of organic and fine materials under S. vulgaris. The range of spatial autocorrelation found in the mass of organic matter on the soil surface was smaller than that of the coverage of S. vulgaris (11.5 m) which corresponded to the canopy patch size of this species, and was longer than the canopy patch size of A. ordosica. The ranges of total C and N, and pH (11.7–15.6 m) were similar to that of S. vulgaris. The range of available P (106.3 m) was comparable to that of the coverage of A. ordosica (86.2 m) considered to be the scale of the distribution of this species. The ranges of silt+clay and exchangeable K, Ca, and Mg (31.0–46.7 m) were not related to plant presence, and were similar to that of topography (43.1 m).     

Experimental infection studies of Pasteurella pneumotropica and V-factor dependent Pasteurellaceae for F344-rnu rats.

To investigate the pathogenicities of P. pneumotropica (Pp) and V-factor dependent Pasteurellaceae (VFDP) in immunodeficient rats, experimental infections of F344-rnu rats were performed using 3 strains (ATCC 35149, CNP 160 and RPZ) of Pp and 4 strains (V6, V7, V8 and V9) of VFDP. Four animals per experimental group were inoculated twice on day 0 and post-inoculation day (PID) 14 with bacterial suspension intranasally. Two animals from each group were sacrificed on PID 60 and 120, and examined. In the animals inoculated with strains of Pp, sneezing was observed in some animals inoculated with strains ATCC 35149 and CNP 160 until PID 31. No clinical signs were observed in other animals. The strains were mainly isolated from the nasal cavity and trachea on PID 60, and the nasal cavity, trachea and lung on PID 120. Inflammation and necrosis of nasal cavity mucosa were observed in all animals inoculated with strains ATCC 35149 and CNP 160 in a histopathologic examination. No histopathological changes were observed in any other animal. In the animals inoculated with strains of VFDP, neither clinical disorder nor histopathological change was observed. The strains were mainly isolated from the trachea on PID 60, and from the trachea and lungs on PID 120. From these results, the pathogenicity of Pp in immunodeficient rats appears to differ by strain, and VFDP appears to be non-pathogenic in immunodeficient rats.     

Highly sensitive quantification of vancomycin in plasma samples using liquid chromatography-tandem mass spectrometry and oral bioavailability in rats

We developed a highly sensitive liquid chromatography-tandem mass spectrometry assay (LC-MS-MS) for a glycopeptide antibacterial drug, vancomycin (VCM), in rat plasma. After precipitating 100 micro l of plasma with 300 micro l of 10% trifluoroacetic acid-methanol (2:1, v/v), the supernatant was diluted with 300 micro l of distilled water and was passed through a filter. LC-MS-MS equipped with electrospray ionization in the positive ion mode used a pair of ions at 725/144 m/z for VCM in the multiple reaction-monitoring mode with a sample injection volume of 20 micro l. The calibration curve had a linear range from 0.01 to 20 micro g/ml when linear least square regression was applied to the concentration versus peak area plot. The drug in the sample was detected within 5 min. Precision, accuracy and limit of quantitation indicated that this method was suitable for the quantitative determination of VCM in rat plasma. Using this method, we defined for the first time that the oral bioavailability of VCM in rats was 0.069%. This method can be applied to basic pharmacokinetic and pharmaceutical studies in rats.     

Biogeochemical and hydrological controls on carbon export from a forested catchment in central Japan

Here we review research on the links between hydrological processes and the biogeochemical environment controlling the dynamics of dissolved organic carbon (DOC) and dissolved inorganic carbon (DIC) in temperate forested catchments. In addition, we present the results of original experiments. The spatial and temporal changes in DIC and DOC concentrations were investigated in tandem with observations of elementary belowground hydrological processes for a forested headwater catchment in central Japan. The soil CO₂ gas concentration, which is the source of DIC, increased with depth. The hydrological characteristics of groundwater also affected the spatial variation of partial pressure of dissolved CO₂ (pCO₂) in groundwater. The temporal variations in the soil CO₂ gas concentration and the pCO₂ values of groundwater suggested that the dynamics of DIC were strongly affected by biological activity. However, the geographical differences in DIC leaching were affected not only by the link between climatological conditions and biological activity, but also by other factors such as geomorphologic conditions. The DOC concentrations decreased with selective removal of hydrophobic acid during vertical infiltration. The major DOC-removal mechanisms were retention of metal-organic complexes to soil solids in the upper mineral soil layer and decomposition of DOC in the lower mineral soil layer. The responses of the DIC and DOC concentrations to changes in discharge during storm events were explained by the spatial variation in the DIC and DOC concentrations. Seasonal variation, which represents a long-term change, in stream water DOC concentrations was affected not only by the temporal variation in DOC concentrations in the topsoil, which may be affected by biological activity, but also by water movement, which transports DOC from the topsoil to stream water. These results indicate that both a biogeochemical approach and a method for evaluating the hydrological effects on carbon dynamics are critical for clarifying the carbon accumulation-and-release processes in forested ecosystems.     

Importance of hydrophobic interaction between a SoxB-type cytochrome c oxidase with its natural substrate cytochrome c-551 and its mutants

Cytochrome c-551, the electron donor of SoxB-type cytochrome c oxidase in thermophilic bacilli, can be over-expressed in Bacillus thermodenitrificans cells by tranformation with pSTEc551. Several mutant cytochromes c-551 were prepared by site-directed mutagenesis to this expression plasmid. Among them, several Lys residues were changed to Ala/Ser, and we found that these mutant cytochromes retained their activity as substrates, although their K(m) values were 0.04-0.12 microM, depending on the site replaced. In contrast, the C19A mutant cytochrome, which was produced in Brevibacillus choshinensis as a secretion protein, lost its activity as a substrate, suggesting that the fatty acyl-glyceryl residue covalently bound to the cysteine residue of the wild-type c-551 plays a very important role in the activity. The importance of the hydrophobic fatty acid residue for the binding of cytochrome c-551 to the oxidase was also shown by the loss of substrate activity in deacylated cytochrome c-551. These results show the importance of the hydrophobic interaction between this cytochrome and SoxB-type oxidase, despite the fact that the importance of an electrostatic interaction between cytochrome c and mitochondrial cytochrome aa(3) oxidase has already been established.     

Stabilization of mast cells by heme oxygenase-1: an anti-inflammatory role

This study examined the role of bilirubin in heme oxygenase (HO)-1-mediated amelioration of mast cell (MC)-elicited inflammatory responses. Pretreatment of rats with an intraperitoneal injection of hemin, an inducer of HO-1, evolved a marked induction of the enzyme in MCs. Intravital videomicroscopy revealed that hemin pretreatment attenuated compound 48/80-elicited degranulation of MCs and resultant leukocyte adhesion in venules. Superfusion with biliverdin or bilirubin, but not with carbon monoxide (CO), another product of the HO reaction, mimicked suppressive actions of the HO-1 induction on both the cell degranulation and leukocyte adhesion elicited by the stimulus, suggesting a requirement of the enzyme reaction to generate bilirubin in the inhibitory mechanisms. Such MC-desensitizing actions of bilirubin were observed in primary-cultured MCs and reproduced irrespective of the choice of stimuli, such as compound 48/80, calcium ionophore, and anti-IgE serum. Furthermore, MC-stabilizing effects of HO-1 were reproduced by the gene transfection of the enzyme into mastocytoma cell line RBL2H3. These results suggest that bilirubin generated through HO-1 serves as an anti-inflammatory substance that desensitizes MCs and ameliorates leukocyte recruitment.     

STUDIES ON THE METABOLISM OF A SULFUR-OXIDIZING BACTERIUM: I. OXIDATION OF SULFUR

1. Thiobacillus thiooxidans, isolated in our laboratory, was foundto oxidize sulfur, but not thiosulfate. Tetrathionate is alsooxidized slightly. Its ability to oxidize sulfur is inactivatedeven by such a mild treatment as keeping the cells in a frozenstate.
2. Inhibitory action of alcohols on the sulfur oxidationincreasesas the length of carbon chain of alcohols increases.Carboxylicacids do not inhibit the sulfur oxidation at pH abovetheirpK, while they strongly inhibit the reaction at pH belowthepK.
3. The sulfur oxidation is inhibited by cyanide, azide,diethyldithiocarbamateand carbon monoxide, and the inhibitionby carbon monoxide isnot reversed by light. These results suggestthe presence ofmetal enzymes in the sulfur oxidation system.The terminal enzymeof this reaction appears to be differentfrom the usual cytochromeoxidase.

(Received May 13, 1960; )     

Identification procedure for Pasteurella pneumotropica in microbiologic monitoring of laboratory animals.

Discrepancies have been recognized in the identification of Pasteurella pneumotropica between testing laboratories. To determine the causes of the differences and to propose a reliable identification procedure for P. pneumotropica, a working group was organized and 69 isolates identified or suspected as P. pneumotropica were collected from 8 laboratories in Japan. These isolates were examined by colony morphology, Gram-staining, the slide agglutination test using two antisera (ATCC35149 and MaR), two commercially available biochemical test kits (ID test, API20NE) and two primer sets of PCR tests (Wang PCR, CIEA PCR). The 69 isolates and two reference strains were divided into 10 groups by test results. No single procedure for P. pneumotropica identification was found. Among tested isolates, large differences were not observed by colony morphology and Gram-straining except for colony colors that depended on their biotypes. Sixty-eight out of 69 isolates were positive by the slide agglutination test using two antisera except for one isolate that tested with one antiserum. The ID test identified 61 out of 69 isolates as P. pneumotropica and there was no large difference from the results of CIEA PCR. From these results, we recommend the combination of colony observation, Gram-straining, the slide agglutination tests with two antisera and biochemical test using the ID test for practical and reliable identification of this organism.