Effect of sodium acetate on the adhesion to porcine gastric mucin in a Lactococcus lactis strain grown on fructose

The association of lactic acid bacteria with mucosal surfaces plays important roles in the beneficial effects of these bacteria on human health, such as colonization of the gastrointestinal tract for pathogen antagonism. Previously, we found that the adhesion of Lactococcus lactis 7‐1 to porcine gastric mucin was higher with fructose than with lactose, galactose or xylose as the carbon source. In this study, we examined the effect of growth conditions on the adhesion of strain 7‐1 grown on fructose. Medium components affect the adhesion: the adhesion of strain 7‐1 grown with sodium acetate was higher than that without it. The enhancement of adhesion by sodium acetate was not observed under aerobic conditions. Cellular properties grown with or without sodium acetate were characterized: strain 7‐1 grown with sodium acetate had similar sugar contents, and different fatty acid composition to those grown without it. Strain 7‐1 grown with sodium acetate showed significantly lower cell yield and significantly higher hydrophobicity than those grown without it, which is associated with higher adhesion. Fructose and sodium acetate are frequently used in the food industry; this study may reveal a simple way to enhance the adhesion of lactic acid bacteria by growing them with these substances.     

Development of furnished cages re‐using conventional cages for laying hens: Behaviour,physical condition and productivity

Furnished cages for laying hens have advantages in allowing normal behaviors and maintaining productivity. As the cost of introduction is a barrier for farms, we developed furnished cages that re‐use conventional cages. To determine the minimum and functional cage design, we compared six designs, combinations of two floor designs (artificial turf or wire cage floor) and three screening designs in the integrated area (no screening, one entrance side or four sides). In total, 144 hens were used, and we measured behavior, physical condition and productivity. Comparing the floors, the percentages of hens performing dust‐bathing and laying eggs in the integrated area were higher in cages with turf than wire floor (P < 0.05 for both). Comparing the screening, dust‐bathing, litter‐exploring and active behavior tended to be more frequent in cages with the integrated area screened on one side than four sides. Feather damage was lower in cages with the integrated area screened on one side than with no screening (P < 0.05). These results suggest that the cage design with an integrated area with artificial turf floor, screened on one side, was effective for furnished cages that re‐use conventional cages.     

不同地下水位胡杨蒸腾速率与叶水势的变化分析

本文研究了不同地下水位胡杨蒸腾速率、叶水势及蒸腾速率的季节变化等水分生理特性,结果表明:地下水位影响胡杨蒸腾作用,同时土壤水分状况也影响蒸腾速率,当地下水位在2.28~2.43 m时,胡杨蒸腾速率变化幅度大,蒸腾速率的日最大值为7.50 mmol.m-2s-1,日均值为4.73 mmol.m-2s-1,对应叶水势的日最低值为-3.55(Mpa)和日均值为-1.94(Mpa);地下水位2.91~3.33 m时,叶水势的日最低值为-2.61(Mpa)和日均值为-1.62(Mpa)。胡杨蒸腾速率季节变化在7月份达到最大值,靠近河岸胡杨蒸腾速率变化较远离河岸胡杨大,表明地下水位低时,胡杨保持较高的水势就能从环境中吸收到水分来满足其正常生长。     

Photosynthetic induction responses to variable light under field conditions in three species grown in the gap and understory of a Fagus crenata forest

Photosynthetic induction responses to abrupt increases in photon flux density (PFD) to 800 and 1500 &mgr;mol m(-2) s(-1) from either darkness or 100 &mgr;mol m(-2) s(-1) were examined in situ in leaves of Fagus crenata Blume, Daphniphyllum humile Maxim., and Acer rufinerve Siebold & Zucc. growing in a gap and the understory of an F. crenata forest. Among the species studied, F. crenata exhibited the highest assimilation rate (A(100)), stomatal conductance (g(s100)) at the background PFD of 100 &mgr;mol m(-2) s(-1), and A(100)/A(max) (A(max) = maximum assimilation rate), in both the gap and the understory. Time required for full induction depended on both background PFD and maximum PFD. The induction period was 2-4-fold shorter at a background PFD of 100 &mgr;mol m(-2) s(-1) than in darkness. For the three understory species, time required to full induction was 2-3-fold longer when irradiance was increased from darkness to 800 &mgr;mol m(-2) s(-1) than when irradiance was increased from darkness to 1500 &mgr;mol m(-2) s(-1). Acer rufinerve showed higher initial stomatal conductance (g(s0)) and a shorter induction period in the understory than in the gap. Fagus crenata exhibited a similar g(s0) and induction period in both habitats. Daphniphyllum humile demonstrated lower g(s0) and a longer induction period in the understory than in the gap. These findings indicate that initial stomatal conductance is closely correlated with the photosynthetic induction response. We conclude that the photosynthetic induction response is affected by the light conditions experienced by plants before the sudden increase in irradiance and by the extent of the increase in irradiance.     

Individual-based measurement and analysis of root system development: case studies for <Emphasis Type="Italic">Larix gmelinii</Emphasis> trees growing on the permafrost region in Siberia

We present results of individual-based root system measurement and analysis applied for Larix gmelinii trees growing on the continuous permafrost region of central Siberia. The data of root excavation taken from the three stands were used for the analyses; young (26 years old), mature (105 years old), and uneven-aged over-mature stand (220 years old). In this article, we highlight two topics: (1) factors affecting spatio-temporal pattern of root system development, and (2) interactions between aboveground (i.e., crown) and belowground (i.e., root) competition. For the first topic, the detailed observation of lateral roots was applied to one sample tree of the overmature stand. The tree constructed a superficial (<30 cm in depth) and rather asymmetric root system, and each lateral root expanded mainly into elevated mounds rather than depressed troughs. This indicated that spatial development of an individual root system was largely affected by microtopography (i.e., earth hummocks). For these lateral roots, elongation growth curves were reconstructed using annual-ring data, and annual growth rates and patterns were compared among them. The comparison suggested that temporal root system development is associated with differences in carbon allocation among the lateral roots. For the second topic, we examined relationships between individual crown projection area (CA) and horizontal rooting area (RA) for the sample trees of each stand. RA was almost equal to CA in the young stand, while RA was much larger (three or four times) than CA in the mature and overmature stands. Two measures of stand-level space occupation, crown area index (aboveground: CAI; sum of CAs per unit land area) and rooting area index (belowground: RAI; sum of RAs), were estimated in each stand. The estimates of RAI (1.3–1.8 m² m₋₂) exceeded unity in all stands. In contrast, CAI exceeded unity (1.3 m² m₋₂) only in the young stand, and was much smaller (<0.3 m² m₋₂) in the two older stands. These between-stand differences in RAI–CAI relationships suggest that intertree competition for both aboveground and belowground spaces occurred in the young stand, but only belowground competition still occurred in the two older stands. Based on this finding, we hypothesized that competition below the ground may become predominant as a stand ages in L. gmelinii forests. Methodological limitations of our analysis are also discussed, especially for the analysis using the two indices of space occupation (CAI, RAI).     

Effects of oxidative stress caused by tert-butylhydroquinone on cytotoxicity in MDCK Cells

Antioxidant and oxidative stress effects of prooxidants are generally dose-dependent, and these effects depend on the prooxidant species and cell type. However, the cellular response to oxidant challenge is a complicated interplay of events involving cellular expression of phase II detoxification enzymes and cellular metal metabolism. This study demonstrates the effect of tert-butylhydroquinone (t-BHQ)-induced oxidative stress on MDCK cells. Cell toxicity tests were carried out using the crystal violet (CV) assay with the following prooxidants: t-BHQ, diethyl maleate (DEM), hydrogen peroxide (H(2)O(2)), diquat (DQ) and β-naphthoflavone (β-NF). Except for β-NF, these prooxidants showed dose-dependent cytotoxicity besides the most potent t-BHQ cytotoxicity. Only t-BHQ and DEM caused significant time-dependent expression of ferritin protein as an antioxidant, which segregates Fe(2+), causing the Fenton reaction. t-BHQ and DEM increased formation of lipid peroxidation, but DQ showed a tendency to decrease lipid peroxidation levels. In XTT assay, even when substantial cell death was observed in the CV assay, t-BHQ appeared to increase cell viability by enhancing XTT reduction, likely through the production of NADPH. Although curcumin, which induces cytoprotective phase II enzymes and chelates metal irons, decreased cell viability, it inhibited t-BHQ cytotoxicity. These results indicate that t-BHQ exhibits strong cytotoxicity against MDCK cells, an effect mitigated by curcumin, and that t-BHQ-induced oxidative stress activates the pentose phosphate pathway.     

Leukotoxin family genes in Staphylococcus aureus isolated from domestic animals and prevalence of lukM-lukF-PV genes by bacteriophages in bovine isolates

Leukotoxin family genes in Staphylococcus aureus isolated from domestic animals were examined by polymerase chain reaction. LukS and lukF genes were detected in all 48 avian and 72 porcine isolates of S. aureus. LukE and lukD genes, located in a putative staphylococcal pathogenicity island (Sapln3/Saplm3), were recognized in 44 (91.7%) of 48 avian isolates, but these genes were not detected in porcine isolates. In 297 bovine isolates collected from mastitic cow's milk and bulk milk from dairy farms in two regions, lukM and lukF-PV(P83) genes in addition to lukS-lukF and lukE-lukD genes were detected in 100 (62.5%) of the 160 isolates from Ishikawa and in118 (86.1%) of the 137 isolates from Hokkaido. When the lysogeny of S. aureus bovine isolates was examined by treatment with mitomycin C, clearing of the culture due to cell lysis was observed in 34 (91.9%) of 37 lukM-lukF-PV(P83) genes--positive isolates. In addition, we isolated a novel lukM-lukF-PV(P83)-carrying (designated phiLukM), and revealed that the lukM-lukF-PV(P83) genes were located very close to an amidase gene on the temperate phage genomes. These results suggest horizontal transmission of lukM-lukF-PV(P83) genes by temperate bacteriophages in S. aureus of bovine origin.     

Subcellular localization of the protein encoded by virulence gene <Emphasis Type="Italic">psvA</Emphasis> of <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">eriobotryae</Emphasis>

The plasmid-encoded virulence gene psvA was previously isolated from Pseudomonas syringae pv. eriobotryae and sequenced. The deduced protein of the psvA gene had no significant similarity to any other protein sequences in the database. To gain a better understanding of the function of the PsvA protein its subcellular localization was examined. To localize the PsvA protein within the bacteria, the cells were fractionated into cytoplasmic, inner membrane, and outer membrane components. The cell fractions and culture supernatant were analyzed by immunoblotting. The PsvA protein was predominantly detected in the outer membrane fraction. Immunoelectron microscopy also showed that the PsvA protein was located in the outer membrane.     

A 39 kDa protein mediates adhesion of avian Pasteurella multocida to chicken embryo fibroblast cells

To clarify the role of avian Pasteurella multocida capsule in pathogenesis, adhesion of capsulated strains P-1059, X-73 and Pm-18, and noncapsulated strains P-1059B, Pm-1 and Pm-3 to chicken embryo fibroblast (CEF) cells was compared. Number of adherent organisms of the capsulated strains to CEF cells were approximately three times as much as noncapsulated strains indicating that adhesive properties were enhanced by the presence of bacterial capsule. Pretreatments of the bacterial cells with heat, trypsin, or with antiserum caused a marked decrease in adhesion of capsulated strain P-1059 and its noncapsulated variant P-1059B. However, depolymerization of capsular hyaluronic acid with high dose of hyaluronidase enhanced adhesion of these strains. Combined treatments of the bacterial cells with both hyaluronidase and trypsin significantly (P < 0.05) inhibited the adherence of strain P-1059 as compared to the treatment only with trypsin, but strain P-1059B was not affected. SDS-PAGE profiles of crude capsular extract (CCE) prepared from capsulated strain P-1059 and its noncapsulated variant P-1059B grown on dextrose starch agar (DSA) plates by heating at 56 degrees C in a 2.5% NaCl solution demonstrated eight protein bands of 28, 34, 36, 39, 52, 56, 63 and 93 kDa. The 28, 34 and 36 kDa proteins were commonly major for both strains, and the 39 kDa protein was major only for strain P-1059 but poor in strain P-1059B. Outer membrane protein (OMP) profiles were identical with a major protein at 34 kDa and four minor proteins between the two strains. The adhesion of strain P-1059 and strain P-1059B to CEF cells was inhibited significantly (P < 0.01) by treatment with rabbit antisera against P-1059, P-1059B, CCE or 39 kDa protein of strain P-1059 as compared to the treatment with either PBS or with normal rabbit serum. These results indicated that an antigenic 39 kDa protein in the capsule may be responsible for adhesion of avian P. multocida type A strains to CEF cells as a virulence factor.