Neuraminidase production by Erysipelothrix rhusiopathiae

In order to characterise neuraminidase activity by Erysipelothrix, 85 isolates of Erysipelothrix spp. from a variety of sources including human clinical, marine and terrestrial animals, and the environment were investigated for neuraminidase production. Neuraminidase activity was detected by a peanut lectin haemagglutination method. The effects of media, incubation conditions and pH on the production and activity of neuraminidase were also investigated. Enzyme activity was detected only in the supernatants of the isolates of Erysipelothrix rhusiopathiae which had been incubated in cooked meat broth and Todd Hewitt broth supplemented with horse serum after 16 and 36 h incubation at 37 degrees C. The maximum titres were reached at 40 h in cooked meat broth and 56 h in Todd Hewitt broth supplemented with horse serum. All 58 isolates and the type strain (ATCC 19414) of E. rhusiopathiae produced detectable neuraminidase activity with titres between 10 and 320. The optimal pH for the enzyme activity varied among the isolates with a pH between 6.0 and 7.0 covering the highest enzyme activity of the most. There was no statistically significant difference in the level of neuraminidase activity between isolates from different sources (p > 0.05). Neuraminidase activity was not detected in the non-pathogenic Erysipelothrix spp. such as E. tonsillarum. Neuraminidase was detected only in E. rhusiopathiae suggesting its possible role as a virulence factor. Enzyme production and activity were medium and pH dependent. The peanut lectin haemagglutination assay is a simple, rapid and sensitive method and is particularly useful for the analysis of multiple samples.     

Effects of Phosphate Type on the Quality of Vacuum-Tumbled Catfish Fillets

The objective of this study was to determine the effects of various agglomerated phosphate blends on the quality of vacuum-tumbled catfish fillets. Catfish fillets were tumbled with a brine solution at 15% over initial, raw weight prior to tray-packing and storage at 4°C for 10 days. Fillets were evaluated for protein exudate, tumbling yield, color, pH, cooking loss, tenderness, purge loss, and shelf life. A specific blend of agglomerated sodium phosphates (AGSP) that contains mono-, tri-, and polyphosphates had significantly less protein (p < 0.05) exudate and significantly higher pH (p < 0.05) than other treatments. All phosphate treatments significantly increased (p < 0.05) tenderness and significantly decreased (p < 0.05) purge loss, but agglomerated phosphate blends significantly decreased (p < 0.05) cooking loss and yellowness. Psychrotrophic plate counts for all phosphate treatments were similar to the control at each storage time. All phosphate treatments improved the yield and quality of catfish fillets, but the use of AGSP may optimize quality attributes.     

Comparison of the standard predictive equation for calculation of resting energy expenditure with indirect calorimetry in hospitalized and healthy dogs

OBJECTIVE: To determine the level of clinical agreement between 2 methods for the measurement of resting energy expenditure (REE). DESIGN: Prospective case series. ANIMALS: 77 dogs. PROCEDURE: Oxygen consumption (Vo2) and CO2 production (Vco2) were measured with an open-flow indirect calorimeter in healthy (n = 10) and ill (67) dogs. Measurements were collected at 3 time periods on 2 days. The Vo2 and the Vco2 measurements were then used to calculate the REE values. RESULTS: Mean values of measured (MREE) and predicted (PREE) REEs in healthy dogs and a dog with medical illnesses or trauma were not significantly different. There was a significant difference on day 2 between the MREE and PREE in the group of dogs recovering from major surgery. More importantly, there was significant variation between the PREE and MREE on an individual-dog basis. The PREE only agreed to within +/- 20% of the MREE in 51% to 57% of the dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The level of agreement between these two methods for determining the 24-hour REE was poor in individual dogs. The level of disagreement between the 2 methods indicates that these methods may not be used interchangeably in a clinical setting. Measurement of REE by use of indirect calorimetry may be the only reliable method of determining REE in an individual ill or healthy dog.     

Catch per unit effort standardization of the eastern Bering Sea walleye pollock (Theragra chalcogramma) fleet

A general linear model (GLM) was used to standardize catch per unit effort (CPUE) data for Alaska walleye pollock (Theragra chalcogramma) from the Bering Sea fleet for the years 1995–1999. Data were stratified temporally by year and season and spatially by area using either Alaska Department of Fish and Game (ADF&G) or National Marine Fisheries Service (NMFS) reporting areas. Four factors were used: vessel identification (ID) number, vessel speed, percentage of pollock by weight in the haul (a measure of targeting), and whether most of the haul took place before or after sunset. At least 29 combinations of main effects, quadratic covariates, and interactions were tested for each year/area/season stratum. GLM models explained from 31 to 48% of the total sums of squares. Vessel identification number was included in all models and explained the most variability. Of the remaining factors, the square of the percentage of pollock in the haul was included in most models, following an F-test to determine parsimony. Analysis of the vessel identification number coefficients indicated that larger vessels tended to have higher CPUEs; and that this relationship differed between dedicated catcher vessels and offshore catcher processors. Coefficient estimates and response surfaces generally indicated increased CPUEs with the percentage of pollock in the haul and showed mixed results with vessel speed. The vessel identification number incorporated most vessel characteristics, leaving vessel speed primarily as a fitting variable with less biological meaning. The year/area/season stratification procedure was found to be necessary due to the unbalanced design, which otherwise would have factor levels with no data in a large combined model. In addition, the stratification procedure reduced the variability in CPUE substantially.     

Counting low-copy number proteins in a single cell

We have designed a microfluidic device in which we can manipulate, lyse, label, separate, and quantify the protein contents of a single cell using single-molecule fluorescence counting. Generic labeling of proteins is achieved through fluorescent-antibody binding. The use of cylindrical optics enables high-efficiency (approximately 60%) counting of molecules in micrometer-sized channels. We used this microfluidic device to quantify beta2 adrenergic receptors expressed in insect cells (SF9). We also analyzed phycobiliprotein contents in individual cyanobacterial cells (Synechococcus sp. PCC 7942) and observed marked differences in the levels of specific complexes in cell populations that were grown under nitrogen-depleted conditions.     

The incorporation of environmentally derived 87Sr/86Sr and Sr/Ca in early otolith formation of Chinook salmon

Effective species management often requires understanding patterns of movement and habitat use. A common approach in identifying where individuals reside relies upon chemical tracers from the environment that are incorporated into an individual's tissues. For fish, isotopes in their otoliths, specifically the portion of their otolith formed during their larval stage, have been used to identify the natal origin. Complicating this work, however, is the fact that during this life stage, there is a shift in the source of isotopes deposited onto the growing otolith from maternally to environmentally derived. The objective of this study was to identify the portion of the otolith representing this transition to environmentally derived isotopes so as to accurately investigate questions of natal origin for a threatened population of fall Chinook salmon (Oncorhynchus tshawytscha). We exposed developing larvae to four treatments that differed in terms of their water strontium isotope ratio (⁸⁷Sr/⁸⁶Sr) and used change-point analysis of otolith ⁸⁷Sr/⁸⁶Sr and strontium to calcium ratio (Sr/Ca) to identify the otolith radius corresponding to the transition to environmentally derived isotopes. Our results indicated this transition occurred, on average, at 132 μm (⁸⁷Sr/⁸⁶Sr; ±50 μm standard deviation) and 127 μm (Sr/Ca; ±29 μm) from the otolith core, which corresponded to the developmental time between hatching and exogenous feeding. A substantial proportion of our otoliths (i.e., 61%) did not show convergence between otolith and water ⁸⁷Sr/⁸⁶Sr by the end of the 113-day experiment, which was likely due to the dietary contribution of marine-based feed. Therefore, we were unable to recommend an otolith radius to target for the purposes of reconstructing natal origin apart from being beyond approximately 130 μm.     

Amphibian oncology.

Spontaneous neoplasia is rare in all three orders of Amphibia.Tumors are documented in most major organ systems, and some have various underlying etiologies, including viral infection, environmental contaminants, and genetic predisposition. Currently,treatment options are limited to removal of the predisposing condition(s), palliative care, surgical excision, and, when necessary,humane euthanasia may be elected. Neoplasia must be distinguished from common infectious, nonneoplastic conditions that can negatively impact population health. This article is a review of the more common types of neoplasia in amphibians, and includes clinically relevant information, such as biologic behavior,anatomy, associated etiologies, major differential diagnoses, and clinical management.