Use of an optical biosensor with a silicone-immobilized enzyme to determine plasma total cholesterol concentrations in fish

Decreased plasma total cholesterol concentrations are a useful indicator in fish of reduced resistance to bacterial infection. Taking advantage of the hydrophobic properties of silicone rubber, we developed an optical enzyme sensor system for measuring plasma cholesterol concentrations in fish. The system was constructed using an immobilized enzyme membrane, optical oxygen probe, flow system, and personal computer. The enzyme membrane was prepared from cholesterol esterase, cholesterol oxidase, and silicone rubber. The calibration curves were linear in the range of 55–85 mg dl⁻¹ for yellowtail and 20–90 mg dl⁻¹ for rainbow trout. Assays could be completed within 3 min, and the sensor response was stabilized by the hydrophobic properties of silicone rubber. Storage of immobilized enzyme membranes at 5°C permitted stable measurements for 58 days. The sensor system was applied to determine plasma total cholesterol concentrations in fish. Good correlations were obtained between results obtained using the sensor and results obtained using conventional methods (correlation coefficients: yellowtail 0.9751, rainbow trout 0.9947). Our method required much less time than conventional cholesterol assays and can be used economically for continuous determination of plasma total cholesterol in fish.     

Rapid enumeration of bacteria grown on surimi-based products by flow cytometry

ABSTRACT: A rapid method to enumerate bacteria adhered on a surimi-based product (kamaboko) by flow cytometry (FCM) is described. To remove Escherichia coli cells from the surface of kamaboko, ultrasonic energy was used. Almost all cells can be removed from kamaboko in 3 min with ultrasonic treatment. Because the sample might contain various non-bacterial particles such as food additives and debris of products, propidium iodide was used to discriminate bacterial cells from non-bacterial particles. Fluorescence scattergrams could distinguish bacteria from the particles, and the FCM method could be used to enumerate bacteria adhered on the surface of kamaboko during storage. Cell numbers determined by FCM paralleled well with those measured using a traditional colony counting method in the range of 10⁴–10⁸ cells/g. The FCM assay could enumerate cells within 1 min and the total assay time, including sample preparation, was less than 30 min.     

Fatty acid‐binding protein expression in the gastrointestinal tract of calves and cows

Fatty acid‐binding protein (FABP) has high affinity for long‐chain fatty acids and appears to participate in the metabolism and intracellular transport of lipids. Liver‐ and intestinal‐type FABP (L‐FABP and I‐FABP, respectively) are expressed in the small intestine. However, in the gastrointestinal tract of ruminants, expression and localization of FABPs are unknown. In this study, we investigated the expression of I‐FABP and L‐FABP in the gastrointestinal tract of cattle. I‐ and L‐FABP had higher messenger RNA (mRNA) and protein expression levels in the duodenum and jejunum relatively to other gastrointestinal regions in both calves and cows. Furthermore, L‐FABP mRNA and protein expression were high in the colon. Both these protein types were confirmed to be in the cytosol of jejunal epithelial cells, where they were found in the villi rather than in the crypts. We concluded that duodenal and jejunal FABPs might be involved in the metabolism of fatty acids mainly in epithelial cells in cattle.     

Effects of sterilization on the chemical forms of heavy metals in soils

We used sequential extraction to investigate changes in the amounts of six chemical forms of manganese, cobalt and cadmium in soil samples after chloroform fumigation. The six forms were designated as follows: exchangeable, dilute-acid-soluble, manganese-oxide-occluded, organically bound, iron-oxide-occluded and residual. For all three metals, the decreases in the amounts of manganese-oxide-occluded forms were equivalent to the sum of the increases in the amounts of exchangeable and dilute-acid-soluble forms. The amounts of the other three forms did not change significantly after fumigation. These results indicate that some of the cobalt and cadmium in the manganese oxides was converted into exchangeable and dilute-acid-soluble forms, which suggests that soil sterilization may increase the availability of these heavy metals to plants.     

Seasonal Variations of Nitrogen Isotopic Ratios of Ammonium and Nitrate in Precipitations Collected in the Yahiko–Kakuda Mountains Area in Niigata Prefecture,Japan

Water, Air, & Soil Pollution - Precipitation was collected from May 15, 2001 to November 18, 2002, at the mountain top (620 m a.s.l.) and mountain foot (47 m a.s.l.) of the...     

Development of an air-injection press for preventing blowout of particleboard IV: effects of air-injection conditions on board performance and formaldehyde emission

An air-injection press, which has holes punched in the heating plates, injects high-pressure air through the holes of one plate into particleboard and discharges the air through the other plate during press heating. The press can manufacture particleboard from high-moisture particles by controlling blowout of the boards. In this study, the optimum diameter and spacing of the air-injection holes and the effects of pre- and post-pressing were investigated. An optimum hole diameter was not found for the modulus of rupture and thickness swelling for a spacing of either 25 or 50 mm. In terms of internal bond strength, the optimum diameter of the holes arranged at a spacing of 25 mm was 1 mm, but the internal bond strength was not changed by the diameter of holes spaced 50 mm apart. Air injection under all hole conditions reduced the formaldehyde emission from the board. Pre-pressing was tested for further increase in the modulus of rupture and internal bond strength, but was found to have no effect. More efficient use of the air-injection press was achieved by injecting air from the early stages of pressing.     

Performance of particleboard manufactured using air-injection press I: effects of air-injection press on preventing blowout of board manufactured from low-moisture particles

An air-injection press (AIP) was developed to prevent accidental blowouts of boards during production. In this study, the effects of the AIP on preventing blowouts were investigated by artificially creating a blowout-prone condition, and the press was shown to be effective in preventing blowouts. The modulus of rupture of the boards was almost constant irrespective of pressing time. Longer pressing time resulted in higher internal bond strength when pressed at 170 °C. The thickness swelling of the boards pressed at 170 or 190 °C was almost uniform irrespective of pressing time, and the manufactured boards showed performance similar to those manufactured with an ordinary press. The AIP prevented blowouts sufficiently even when the pressure of the injected air was reduced, and this reduction did not adversely decrease the performance of the boards. Air injection reduced formaldehyde emissions from the board.     

Factors associated with antimicrobial-resistant Escherichia coli in zoo animals

Factors associated with the carriage of antimicrobial-resistant Escherichia coli isolates were analysed among zoo animals. An association was observed between selection of amoxicillin as the first-line therapy and a significantly higher percentage of resistance to ampicillin (54.5%) from 11 animals treated with antimicrobials, compared with isolates from 32 untreated animals (9.4%). In addition, the percentage resistance to kanamycin (36.4%), gentamicin (27.3%), trimethoprim (27.3%) and tetracycline (63.6%) from 11 treated animals was significantly higher than those from 32 untreated animals (3.1%, 3.1%, 3.1% and 25%, respectively), although these antimicrobials were rarely used. All kanamycin-, gentamicin- and trimethoprim-resistant isolates and more than half of the tetracycline-resistant isolates from treated animals were also resistant to ampicillin. Co-resistance to other antimicrobials with ampicillin was suggested to contribute to an increasing of resistance towards antimicrobials that were rarely administered. The present investigation revealed an association of antimicrobial treatment with the spread of antimicrobial-resistant bacteria among zoo animals.     

Development of AFLP-derived SCAR markers associated with resistance to two races of southern root-knot nematode in sweetpotato

The southern root-knot nematode (SRKN) Meloidogyne incognita severely damages yield and quality in sweetpotato production, and host plant resistance is one of the primary options for SRKN control. Segregation of F₁ progeny resistant and susceptible to the SP1 and SP2 races of SRKN suggested that the race-specific resistance of the sweetpotato cultivar ??Hi-Starch?? is mostly controlled by single genes and that the genes for resistance against each race are closely located. Bulked segregant analysis and subsequent analysis of 86 F₁ progeny plants identified nine amplified fragment-length polymorphism markers associated with SRKN resistance and a single linkage map consisting of seven of these markers. Quantitative trait locus (QTL) analysis using the segregating resistance data of the F₁ progeny allowed mapping of both a locus with a large effect on resistance to the SRKN race SP1 and another affecting resistance to SP2 to the region around E33M53_090 that was designated as qRmi(t). Two AFLP markers in the vicinity of qRmi(t), E33M53_090 and E41M32_206, were converted to locus-specific sequence-characterized amplified region markers based on their internal and adjacent DNA sequences. These markers might be useful for marker-assisted selection of SRKN resistance in sweetpotato breeding and as a first step to map-based cloning of the responsible QTL(s).     

Seasonal variations in leaf growth of <Emphasis Type="Italic">Cymodocea serrulata</Emphasis> in subtropical seagrass meadows

We estimated the leaf growth of the tropical–subtropical seagrass species Cymodocea serrulata using the conventional leaf-marking method in a subtropical seagrass bed on Ishigaki Island, southern Japan, which is near the northern distributional limit of this species. This survey was conducted over an annual cycle at two sites. The deep site was located in a dense meadow in the subtidal zone. The shallow site was located around the lowest part of the intertidal zone, forming a tide pool during ebb tides of the spring tide, which was considered to approximate the upper depth limit of C. serrulata. The relationships between temperature and shoot weight, and the number of leaves per shoot were significantly influenced by site, resulting in a lower biomass in the shallow site. Leaf growth and shoot length increased with temperature without any spatial variation. Leaf growth and the number of leaves per shoot particularly declined during periods of low temperature in both sites. These results suggest that leaf growth and phenology of C. serrulata in this subtropical area are largely regulated by temperature.