Lactose utilization byVibrio vulnificus

The ability of wild-type strains ofVibrio vulnificus to utilize lactose as a sole source of carbon and energy and produce acid in lactose-containing media is associated with the appearance of spontaneous lactose-utilizing mutants. These contain increased activities of an enzyme able to hydrolyzeo-nitrophenyl--d-galactoside as well as lactose. This activity is constitutive in some mutants and inducible by both lactose and isopropyl--d-thiogalactoside in others. A limited survey of otherVibrio species indicates thatV. pelagius also can acquire, by mutation, the ability to grow on and make acid from lactose. No immunological cross-reaction was detected between the enzymes fromVibrio and the -galactosidases ofEscherichia coli andKlebsiella.     

Regulation systems of protease and hemolysin production in Vibrio vulnificus

Vibrio vulnificus, a gram‐negative halophilic estuarine bacterium, is an opportunistic human pathogen that causes rapidly progressive fatal septicemia and necrotizing wound infection. This species also causes hemorrhagic septicemia called vibriosis in cultured eels. It has been proposed that a range of virulence factors play roles in pathogenesis during human and/or eel infection. Among these factors, a metalloprotease (V. vulnificus protease [VVP]) and a cytolytic toxin (V. vulnificus hemolysin [VVH]) are of significant importance. VVP elicits the characteristic edematous and hemorrhagic skin damage, whereas VVH exhibits powerful hemolytic and cytolytic activities and contributes to bacterial invasion from the intestine to the blood stream. In addition, a few V. vulnificus strains isolated from diseased eels have recently been found to produce a serine protease designated as V. vulnificus serine protease (VvsA) instead of VVP. Similarly to VVP, VvsA may possess various toxic activities such as collagenolytic, cytotoxic and edema‐forming activity. In this review, regulation of V. vulnificus VVP, VVH and VvsA is clarified in terms of expression at the mRNA and protein levels. The explanation is given on the basis of the quorum sensing system, which is dependent on bacterial cell density. In addition, the roles of environmental factors and global regulators, such as histone‐like nucleoid structuring protein, cyclic adeno monophosphate receptor protein, RpoS, HlyU, Fur, ToxRS, AphB and LeuO, in this regulation are outlined. The cumulative impact of these regulatory systems on the pathogenicity of V. vulnificus is here delineated.     

Some properties of Vibrio vulnificus hemolysin

Some properties of hemolysin produced by Vibrio vulnificus were investigated. The hemolysin was heat labile, and the hemolytic activity was inhibited by adding cholesterol or divalent cations. Cholesterol inhibited the temperature-independent hemolysin-binding step, suggesting that cholesterol made up the binding site of the cell membrane, whereas the divalent cations inhibited the temperature-dependent membrane-degradation step. However, the V. vulnificus hemolysin was stable to oxygen and sulfhydryl reagents and was not inactivated by antiserum against streptolysin O, suggesting that the V. vulnificus hemolysin differs from oxygen-labile hemolysins which bind to cholesterol. The V. vulnificus hemolysin seems to be one of the exceptional cholesterol-binding hemolysins.     

Isolation and characterization of hemolysin mutants of Vibrio vulnificus

Abstract The importance of the cytolysin/hemolysin in the virulence of Vibrio vulnificus was investigated using both the naturally occuring virulent and avirulent colony variants and ethylmethane-sulfonate generated mutants. Both virulent and avirulent isogenic morphotypes produced similar amounts of hemolysin. Two mutants deficient in the production of hemolysin and negative for CHO cell activity were characterized and their virulence for mice was examined. Non-hemolytic mutants were found to be as virulent as their parent strain. It is concluded that the hemolysin produced by V. vulnificus is not required for the full virulence of this pathogen.     

Mutations affecting hemolysin production in Listeria monocytogenes located outside the listeriolysin gene

We have investigated the molecular basis of spontaneous mutations leading to non-hemolytic and avirulent variants of the Listeria monocytogenes serotype 1/2a strain NCTC 7973 using Southern hybridization to DNA fragments that harbor the listeriolysin gene (hlyA) and adjacent regions cloned from a L. monocytogenes serotype 1/2a strain. The analysis of such non-hemolytic variants revealed the presence of a deletion of 300 base pairs, located 1.6 kb upstream of an otherwise intact listeriolysin gene. The importance of regions upstream of the hlyA gene in controlling the expression of the listeriolysin gene was further emphasized by the detection of a transposon-derived nonhemolytic mutant in which the transposon had inserted approximately 200 bp upstream of the listeriolysin gene. We conclude that at least two elements, contained within a region encompassing 1.6 kb of sequences upstream of the hlyA gene, may be required for expression of the listeriolysin gene.     

Factors affecting pheromone production in beetles

Pheromone production and/or release by beetles is coordinated with a variety of behavioral, physiological, and environmental factors. To data, two basic mechanisms for the regulation of pheromone biosynthesis in beetles have been proposed. Pheromone biosynthesis may simply be dependent on the availability of biosynthetic precursors. Alternatively, certain stimuli or events may trigger pheromone biosynthesis via juvenile hormone (JH) action. JH may either act directly at the site of pheromone biosynthesis to enhance pheromone production or may act indirectly, through a brain hormone (which might be related to the pheromone biosynthesis activating neuropeptide) or through effects on antennal sensory response. Knowledge of the regulation of the initiation and termination of pheromone biosynthesis is reviewed. Mechanisms by which pheromone stereochemistry is controlled are also discussed. This is an important aspect of pheromone production in Coleoptera, since slight changes in the stereochemistry can completely alter the activity of the molecule. © 1994 Wiley-Liss, Inc.     

Non-thiol-activated property of a cholesterol-binding hemolysin produced by Vibrio vulnificus

Abstract Vibrio vulnificus hemolysin (VVH) and streptolysin O (SLO) are both cholesterol-binding hemolysins. Both hemolysins were inactivated with H₂O₂, but the lost activity of SLO was restored by addition of thiol compounds, whereas that of VVH was not. Moreover, the activity of VVH was lowered by thiol compounds but not by thiolblocking agents, whereas the latter produced a decrease in SLO activity. These results suggest that VVH is not a thiol-activated hemolysin, in spite of its cholesterol-binding property.     

同源四倍体水稻原种制备及影响因素分析

本研究已将从全国各地征集的50份水稻优良种质资源(2n=2x=24)诱变为同源四倍体水稻原种。本文讨论不同处理日期、不同秋水仙碱浓度和处理时间、以及不同品种的抗药性等因素对加倍成功率的影响,结果表明:①不同日期处理水稻幼芽,其加倍成功率不同,4月6日～15日加倍效果最好,加倍成功率达19.4%。②不同浓度秋水仙碱和处理时间其加倍成功率存在一定的差别。0.1%秋水仙碱处理48h 和0.2%处理24h 为佳,其芽恢复率超过58%,加倍成功率超过16%。③不同水稻品种抗药性不同。在所加倍的50个水稻品种中,中作9025抗药性最强,芽恢复率达92.4%,加倍成功率38.7%。盐州14抗药性最差,芽恢复率仅为10.6%。     

Sandwich enzyme-linked immunosorbent assay for Vibrio vulnificus hemolysin to detect V. vulnificus in environmental specimens.

Vibrio vulnificus hemolysin, purified by quantitative isoelectric focusing, was used to prepare rabbit and goat anti-hemolysin. The resulting antibodies were used as capture and detector antibody reagents in a sandwich enzyme-linked immunosorbent assay (ELISA) to detect V. vulnificus in environmental samples. By this technique, 4 laboratory-maintained V. vulnificus strains and 33 environmental V. vulnificus isolates were detected. Also, the technique distinguished five other Vibrio species from V. vulnificus, and when it was used in combination with colistin-polymyxin-cellobiose agar, 31 non-V. vulnificus isolated were excluded. This sandwich ELISA compared favorably with the current Food and Drug Administration standard immunoassay in confirming presumptive V. vulnificus colonies from environmental specimens: oysters, sediment, and seawater. Among 340 presumptive V. vulnificus colonies, the sandwich ELISA detected 95% of the confirmed V. vulnificus colonies. Equally important, the technique correctly distinguished 99% of the non-V. vulnificus colonies. The sandwich ELISA offers time-saving and labor-saving advantages over the currently accepted immunoassay.     

Vibrio vulnificus hemolysin dilates rat thoracic aorta by activating guanylate cyclase

Hemolysin produced by Vibrio vulniflcus caused hypotension and tachycardia in rats and dilated rat thoracic aorta. Hemolysin-induced vasodilatation of the aorta was not affected by Nω-nitro-L-arginine methyl ester and aminoguanidine, NO synthase inhibitors, whereas the vasodilatation was inhibited by LY 83,583, a guanylate cyclase inhibitor. Hemolysin elevated cGMP levels, and the elevation was abolished by LY 83,583. These results suggest that V. vulnificus hemolysin activates guanylate cyclase independently of NO synthase, and the subsequent increase in cGMP levels results in vasodilatation.     

Susceptibility of erythrocytes from several animal species to Vibrio vulnificus hemolysin

The hemolytic activity of Vibrio vulnificus hemolysin (VVH) against erythrocytes from several animal species (sheep, horse, cow, rabbit, chicken) was investigated. VVH was active against erythrocytes from all species, but the amount of VVH causing 50% hemolysis under identical conditions (hemolytic susceptibility to VVH) differed. The degree of 125I-labeled VVH (125I-VVH) binding to each erythrocyte species correlated with the susceptibility of the cells to hemolysis. However, marked differences in the binding ability of 125I-VVH were not observed against liposomes constructed with lipids from each erythrocyte membrane. On the other hand, release of hemoglobin (Hb) differed for each of the erythrocyte species despite administration of approximately the same hemolytic VVH concentration to each species. Furthermore, under hypotonic conditions, the stability of each erythrocyte species varied markedly; the more susceptible the erythrocyte to VVH, the more unstable it was under such conditions. These results, therefore, suggest that the susceptibility of erythrocytes to VVH may be closely associated with the binding ability of VVH and erythrocyte membrane stability.     

Factors affecting ultrasound production in infant rodents

Studies on the motivation for the production of ultrasound in rodents revealed that two principal factors influence this phenomenon in the infants. These have already been reported in full for white mice and partially for Wistar rats and Golden hamsters. The present paper completes the report on all five species that were investigated and makes a general assessment of the findings.     

Factors affecting progeny production of Anaphes iole

Anaphes iole Girault(Hymenoptera: Mymaridae) is a native, solitaryegg parasitoid of Lygus spp.(Heteroptera: Miridae) in North America. Current research is considering factors thatmay optimize the in-vivo rearing of A.iole using Lygus hesperus Knight(Heteroptera: Miridae) as host. The effects ofhost density, day of oviposition and foodpresence, parasitoid age and mate presence onproduction of A. iole progeny weredetermined in this study. After exposingindividual parasitoids to host patches for 24h, the percentage of hosts containing a latestage A. iole pupa wassignificantly greater at a moderate hostdensity (41–70 eggs per patch) than at a lowdensity (10–40 eggs). But, no differenceswere detected between moderate and high density(71–100 eggs) or between high and low densitytreatments. More adult progeny were generatedby females (of variable age; 0–2 d old) onthe first day of oviposition rather than thesecond day, regardless of food presence. Progeny sex ratio was decidedly male-biased onthe second day. Female age (0 d vs 1 d old)had a marginal effect on production in 24 h; 0d old females tended to generate more adultprogeny than 1 d old females. Overall, thisresearch suggests that exposing newly-emerged,unfed, mated A. iole females to amoderate to high host density for 1 to 2 dcould lead to time-efficient production ofadult progeny in an in-vivo rearing system.     

Factors affecting lipase production in Syncephalastrum racemosum

Syncephalastrum racemosum grown as a static culture showed maximum lipase production at 30°C in 2d at pH 8.0. When the medium was supplemented with fructose, maximum production of lipase per unit of growth was achieved, followed by raffinose, sucrose, ribose, galactose, maltose, lactose, mannitol and glucose. Amongst the nitrogen sources tested, corn steep liquor at 8% (v/v) produced maximum enzyme; there was evidence of catabolite repression by glucose when groundnut protein, soybean meal, milk casein or wheat bran were the sources of nitrogen. Calcium, potassium and sodium citrates, each at 0.1% (w/v), increased the yield of lipase.     

Factors affecting goat milk production and quality

Differences between production systems based on grazing and browsing vs. use of harvested feedstuffs in confinement largely depend on specific feedstuffs and plants available and being consumed. Low forage nutrient ingestion should have relatively greater impact on tissue mobilization than milk production in early than later periods of lactation, with a transition to proportionally greater change in milk production in late lactation. However, low body condition at kidding would limit tissue energy mobilization and restrict impact of level of nutrient intake to milk yield and, likewise, tissue mobilization would be less with one vs. two or three milkings per day. As lactation advances after freshening, fat and protein levels decrease with increasing milk yield, and when production declines in mid- to late lactation, fat and protein concentrations increase. Milk production generally peaks at a parity of 3 or 4, thereafter declining slowly. Elevated somatic cell count alone in dairy goats is not a valid indication of mammary infection. Extended lactations offer opportunities to minimize or avoid seasonal fluctuations in milk production and lessen production costs. If differences in performance between suckled and machine-milked dairy goats occur, they may be restricted to or of greater magnitude during the suckling period compared with post-weaning, and differences in milk yield will either be absent or less with one kid compared with greater litter sizes. The magnitude of effects of milking frequency on milk yield is less for goats of low vs. high production potential and with low vs. high diet quality. Likewise, the effect of milking frequency is greater in early and mid-lactation when yield is higher than in late lactation, along with a shorter period of peak production with one vs. two daily milkings. Physical form of the diet can affect production and composition of goat milk, although effects appear of smaller magnitude than in dairy cattle. When tissue is mobilized to support milk production in early lactation, levels of C18:0 and C18:1 cis in milk increase and levels of medium-chain fatty acids decline. Effects of elevated levels of dietary fatty acids on specific long-chain fatty acids in milk and milk products vary with the fatty acid profile of fat sources used.