CRITICAL LEVELS OF LIGHT AND TEMPERATURE REGULATING THE GAMETOGENESIS OF THREE LAMINARIA SPECIES (PHAEOPHYCEAE)1

Gametophytes of three Laminaria species occurring near Helgoland, North Sea, were cultivated 4 wk in a 12:12 LD regime at different temperatures in artificial light fields, and in the sea at different water depths. In the artificial light fields underwater spectral distribution was simulated according to Jerlov water Types 5, 7, 9. Blue light in the simulated light fields amounted to 17, 12 or 4% of total quanta. The rate of vegetative growth did not depend on spectral distribution, was light-saturated at 4–6 W · m^?2, and increased with temperature up to 15 C. L. saccharina (L.) Lamour. exhibited the highest tolerance towards temperature, light and UV. Gametophytes survived 1 wk at 21 C ± 0.1, but not 22 C ± 0.1. Gametophytes of L. hyperborea (Gunn.) Fosl. and L. digitata (Huds.) Lamour. survived 1 wk at 20 C ± 0.1, but not at 21 C ± 0.1. In sunlight, and in the light field of a xenon lamp, 50% of L. saccharina gametophytes were killed by a quantum dose of 50 μEin · cm^?2, and 100% of the plants by 90 μEin · cm^?2. Approximately half of these quantum doses killed the corresponding percent of the other species gametophytes. Appreciably higher quantum doses were survived in visible light, with red being the most detrimental. Fertility depended on a critical quantum dose of blue light which decreased almost exponentially with decreasing temperature. The quantum dose (400–512 nm) required for induction of fertilization of 50% of the female gametophytes (males react similarly) was 90 μEin · cm^?2 at 5 C, 110 μEin · cm^?2 at 10 C, 230 (560 in L. digitata)μEin · cm^?2 at 15 C, and 560 (L. hyperborea) or about 850 (other 2 species) μEin · cm^?2 at 18 C. In the sea the gametophytes survived the dark winter months in the unicellular stage, with almost no vegetative growth of the primary cell, due to lack of light. In early spring the female gametophytes matured in the unicellular, and the males in a few-celled stage at the depth of 2 m, as did the laboratory cultures under conditions inducing maximal fertility.     

Landscape Changes Influence the Occurrence of the Melioidosis Bacterium Burkholderia pseudomallei in Soil in Northern Australia

Background

The soil-dwelling saprophyte bacterium Burkholderia pseudomallei is the cause of melioidosis, a severe disease of humans and animals in southeast Asia and northern Australia. Despite the detection of B. pseudomallei in various soil and water samples from endemic areas, the environmental habitat of B. pseudomallei remains unclear.

Methodology/Principal Findings

We performed a large survey in the Darwin area in tropical Australia and screened 809 soil samples for the presence of these bacteria. B. pseudomallei were detected by using a recently developed and validated protocol involving soil DNA extraction and real-time PCR targeting the B. pseudomallei–specific Type III Secretion System TTS1 gene cluster. Statistical analyses such as multivariable cluster logistic regression and principal component analysis were performed to assess the association of B. pseudomallei with environmental factors. The combination of factors describing the habitat of B. pseudomallei differed between undisturbed sites and environmentally manipulated areas. At undisturbed sites, the occurrence of B. pseudomallei was found to be significantly associated with areas rich in grasses, whereas at environmentally disturbed sites, B. pseudomallei was associated with the presence of livestock animals, lower soil pH and different combinations of soil texture and colour.

Conclusions/Significance

This study contributes to the elucidation of environmental factors influencing the occurrence of B. pseudomallei and raises concerns that B. pseudomallei may spread due to changes in land use.     

DIFFERENTIAL EFFECTS OF PHOTOPHASE IRRADIANCE ON METABOLIC AND URINARY STRESS HORMONE CONCENTRATIONS IN BLIND AND SIGHTED RODENTS

The effects of different photophase irradiance levels on the daily rhythms of energy expenditure (DEE, calculated from oxygen consumption, VO₂) and urinary metabolites of stress hormones in sighted (Microtus socialis) and blind (Spalax ehrenbergi) rodents were compared. Five groups of each species were exposed to different irradiance levels (73, 147, 293, 366, and 498 μW/cm²) under short photoperiod (8L:16D) condition with constant ambient temperature 25?±?2°C for 21 days before assessments. As light intensity increased from 73 μW/cm², both species reduced DEE, especially among M. socialis. Cosinor analysis revealed significant ultradian rhythms in VO₂ of M. socialis with period length being inversely related to irradiance level. Conversely, in S. ehrenbergi, robust 24 h VO₂ rhythms were detected at all irradiances. In M. socialis, significant 24 h rhythms in urinary output of adrenaline were detected only at 293 μW/cm², whereas for cortisol, unambiguous rhythms were detected at 73 and 147 μW/cm². Distinct adrenaline daily rhythms of S. ehrenbergi were observed at 73 and 293 μW/cm², whereas this species exhibited significant rhythms in cortisol at 147 and 293 μW/cm². Changes in photophase irradiance levels affected stress hormone concentrations in a dose-dependent manner. There were significant negative and positive correlations of M. socialis and S. ehrenbergi stress hormones, respectively, with increasing irradiance. Our results indicate photophase light intensity is another environmental factor that can significantly affect entrainment of mammalian daily rhythms. Both low and high irradiance conditions can trigger stress responses, depending on the species' natural habitat. (Author correspondence: zubidat3@013.net.il)     

电解水的抑菌活性及对食品加工表面材料的消毒效果

为了考查应用电解水消除细菌污染的可行性,对氧化电解水的杀菌效果及对食品加工表面材料的消毒效果进行了研究。结果表明,含0.1%NaCl的自来水经7min的电解后所获得的氧化电解水,能在2min内将菌液浓度分别为4.20×106CFU/mL,2.18×106CFU/mL,1.44×106CFU/mL,2.10×106CFU/mL,1.94×106CFU/mL的埃希氏大肠杆菌(Escherichia coliO157:H7)、沙门氏菌(Salmonella enteritidis)、单核细胞增生李斯特菌(Listeria monocytogenes)、摩化摩根菌(Morganella morganii)、副溶血性弧菌(Vibrio parahaemolyticus)几乎全部杀死。另外,对食品加工表面接触材料中的地板砖、不锈钢板、瓷砖进行染菌消毒试验结果表明,含0.1%NaCl的电解水同样能将上述浓度的菌液感染到食品表面接触材料后在5min之内几乎全部将其杀死,是一种理想的食品表面材料消毒剂。     

Inhibitory effects of silver ions on Legionella pneumophila grown on agar, intracellular in Acanthamoeba castellanii and in artificial biofilms

Aims: We undertook a series of experiments to investigate the susceptibility of Legionella pneumophila grown under extracellular and intracellular conditions and other water‐related bacteria to silver ions. Methods and Results: In this study, the antimicrobial effect of silver ions to intra‐ and extra‐cellular grown Legionella bacteria was investigated. The minimal inhibitory concentration (MIC) after 24 h exposure, leading to a 5 log reduction, was c. 64 μg l^?1 AgNO₃ for extracellular grown Legionella and other tested Gram‐positive and Gram‐negative bacteria. In contrast, the MIC for intracellularly grown Legionella was up to 4096 μg l^?1 AgNO₃ after 24 h. Furthermore, the heterotrophic bacteria grown within a biofilm model were killed at a concentration of 4–16 μg l^?1 AgNO₃. In contrast, biofilm‐associated Legionella were less sensitive (MIC 128–512 μg l^?1 AgNO₃). Conclusion: Intracellularly and biofilm‐grown legionellae are less sensitive against silver compared with agar‐grown bacteria. Significance and Impact of the Study: The reduced sensitivity of Legionella grown in amoebae might explain why the effect of silver decontamination requires an extended exposure in field trials.     

Complete killing of Caenorhabditis elegans by Burkholderia pseudomallei is dependent on prolonged direct association with the viable pathogen

Background

Burkholderia pseudomallei is the causative agent of melioidosis, a disease of significant morbidity and mortality in both human and animals in endemic areas. Much remains to be known about the contributions of genotypic variations within the bacteria and the host, and environmental factors that lead to the manifestation of the clinical symptoms of melioidosis.

Methodology/Principal Findings

In this study, we showed that different isolates of B. pseudomallei have divergent ability to kill the soil nematode Caenorhabditis elegans. The rate of nematode killing was also dependent on growth media: B. pseudomallei grown on peptone-glucose media killed C. elegans more rapidly than bacteria grown on the nematode growth media. Filter and bacteria cell-free culture filtrate assays demonstrated that the extent of killing observed is significantly less than that observed in the direct killing assay. Additionally, we showed that B. pseudomallei does not persistently accumulate within the C. elegans gut as brief exposure to B. pseudomallei is not sufficient for C. elegans infection.

Conclusions/Significance

A combination of genetic and environmental factors affects virulence. In addition, we have also demonstrated that a Burkholderia-specific mechanism mediating the pathogenic effect in C. elegans requires proliferating B. pseudomallei to continuously produce toxins to mediate complete killing.     

Influence of soil edaphic factors and their critical limits on seedling emergence of corn (Zea mays L.)

Summary In the present investigation the influence of various soil physical parameters on seeding emergence of corn (Zea mays L.) and their critical limits was studied in order to obtain a proper crop stand. The effect of soil compaction, resulting into various bulk densities on some soil physical properties was also studied. Seedling emergence decreased with increasing bulk density, soil suction, seed placement depth, soil strength and with decreasing oxygen diffusion rates. The critical values for these soil physical parameters were found to be 1.4 g/cm³, 3 atm., 4 cm, 1.0 kg/cm² and 40×10^-8 g O₂/cm²/min bulk density, soil suction, seed placement depth, soil strength and oxygen diffusion rates respectively. Increasing bulk density decreased the soil water content, oxygen diffusion rates and increased the soil strength. re]19751117 Soil Science Department     

Sensitive and Specific Molecular Detection of Burkholderia pseudomallei, the Causative Agent of Melioidosis, in the Soil of Tropical Northern Australia

Burkholderia pseudomallei, the cause of the severe disease melioidosis in humans and animals, is a gram-negative saprophyte living in soil and water of areas of endemicity such as tropical northern Australia and Southeast Asia. Infection occurs mainly by contact with wet contaminated soil. The environmental distribution of B. pseudomallei in northern Australia is still unclear. We developed and evaluated a direct soil B. pseudomallei DNA detection method based on the recently published real-time PCR targeting the B. pseudomallei type III secretion system. The method was evaluated by inoculating different soil types with B. pseudomallei dilution series and by comparing B. pseudomallei detection rate with culture-based detection rate for 104 randomly collected soil samples from the Darwin rural area in northern Australia. We found that direct soil B. pseudomallei DNA detection not only was substantially faster than culture but also proved to be more sensitive with no evident false-positive results. This assay provides a new tool to detect B. pseudomallei in soil samples in a fast and highly sensitive and specific manner and is applicable for large-scale B. pseudomallei environmental screening studies or in outbreak situations. Furthermore, analysis of the 104 collected soil samples revealed a significant association between B. pseudomallei-positive sites and the presence of animals at these locations and also with moist, reddish brown-to-reddish gray soils.     

Chlorine dioxide inactivation of bacterial threat agents

Aims: To evaluate the efficacy of chlorine dioxide (ClO₂) against seven species of bacterial threat (BT) agents in water. Methods and Results: Two strains of Bacillus anthracis spores, Yersinia pestis, Francisella tularensis, Burkholderia pseudomallei, Burkholderia mallei and Brucella species were each inoculated into a ClO₂ solution with an initial concentration of 2·0 (spores only) and 0·25 mg l^?1 (all other bacteria) at pH 7 or 8, 5 or 25°C. At 0·25 mg l^?1 in potable water, six species were inactivated by at least three orders of magnitude within 10 min. Bacillus anthracis spores required up to 7 h at 5°C for the same inactivation with 2·0 mg l^?1 ClO₂. Conclusions: Typical ClO₂ doses used in water treatment facilities would be effective against all bacteria tested except B. anthracis spores that would require up to 7 h with the largest allowable dose of 2 mg l^?1 ClO₂. Other water treatment processes may be required in addition to ClO₂ disinfection for effective spore removal or inactivation. Significance and Impact of Study: The data obtained from this study provide valuable information for water treatment facilities and public health officials in the event that a potable water supply is contaminated with these BT agents.     

Partial identification and copper tolerance of soil bacteria from copper-impregnated bog and from a salt marsh

Abstract

Bacteria from two areas in Wales have been extracted and partially identified. The areas are a copper impregnated bog in the Coed-Y-Brenin Forest with low pH and high carbon and copper content, and the Morfa Harlech salt marshes which have neutral pH and lower copper and carbon. Bacillus spp. was the predominant bacterium in both soils, but there was increased diversity of bacteria in the salt marsh soils. Bacteria from the Cu-rich, Coed-Y-Brenin soils showed higher growth densities in nutrient agar plates containing 10, 30 and 100 μg cm^?3 Cu than bacteria from the salt marsh soils, indicating that bacteria from the Coed-Y-Brenin soils could tolerate higher copper concentrations. Pseudomonas spp. from both study areas grew better in media with added Cu at low levels. The possibility is discussed that the bacteria play a part in the bioavailability of Cu particularly in the copper-rich Coed-Y-Brenin area.     

Effects of environmental factors upon variation in soil respiration of a Zoysia japonica grassland, central Japan

The effects of environmental factors on seasonal and annual variations in soil respiration were examined in the cool temperate Zoysia japonica grassland of Japan. Field measurements of soil respiration were conducted using a closed chamber method with an infrared gas analyzer at monthly intervals in the snow-free seasons from May 2007 to December 2009. There was an exponential relationship between soil respiration and soil temperature, and the soil temperature accounted for 85–86% of seasonal soil respiration variability. Moreover, a positive linear relationship between soil respiration and soil water content was detected in summer (R ² = 0.55, p < 0.001), but not in spring or autumn. Annual soil respiration was estimated at 755, 719, and 1,037 g C m⁻² year⁻¹ in 2007, 2008, and 2009, respectively. These interannual variations in soil respiration might be influenced by the strength of precipitation during rainy seasons and the timing of each snow-melt. Our results suggest that the effects of rainfall and snow-melt events on soil respiration might be important factors to understand carbon dynamics in grassland ecosystem in Japan.     

Evaluation of Molecular Methods To Improve the Detection of Burkholderia pseudomallei in Soil and Water Samples from Laos

Burkholderia pseudomallei is the cause of melioidosis, a severe and potentially fatal disease of humans and animals. It is endemic in northern Australia and Southeast Asia and is found in soil and surface water. The environmental distribution of B. pseudomallei worldwide and within countries where it is endemic, such as the Lao People''s Democratic Republic (Laos), remains unclear. However, this knowledge is important to our understanding of the ecology and epidemiology of B. pseudomallei and to facilitate public health interventions. Sensitive and specific methods to detect B. pseudomallei in environmental samples are therefore needed. The aim of this study was to compare molecular and culture-based methods for the detection of B. pseudomallei in soil and surface water in order to identify the optimal approach for future environmental studies in Laos. Molecular detection by quantitative real-time PCR (qPCR) was attempted after DNA extraction directly from soil or water samples or after an overnight enrichment step. The positivity rates obtained by qPCR were compared to those obtained by different culture techniques. The rate of detection from soil samples by qPCR following culture enrichment was significantly higher (84/100) than that by individual culture methods and all culture methods combined (44/100; P < 0.001). Similarly, qPCR following enrichment was the most sensitive method for filtered river water compared with the sensitivity of the individual methods and all individual methods combined. In conclusion, molecular detection following an enrichment step has proven to be a sensitive and reliable approach for B. pseudomallei detection in Lao environmental samples and is recommended as the preferred method for future surveys.     

模拟喀斯特不同土壤生境下黑麦草对水分胁迫的生长和光合生理响应

以多年生草本黑麦草为对象,模拟喀斯特生境土壤特征设置浅而宽(Shallow and wide, SW:30×30×5 cm~3)和深而窄(Deep and narrow, DN:10×10×45 cm~3)两种土壤容器,以正常降水量为对照供水(W100%),设减水50%(W50%)和减水70%(W30%)共3种水分处理进行盆栽实验,探究了两种不同土壤生境中土壤水分变化对黑麦草生长及光合生理的影响,以进一步理解喀斯特地区植物的适应对策。结果显示:(1)SW生境对水分变化敏感,随供水减少土壤含水量显著下降。轻度减水下植物叶面积增大,光合速率提高,地上部分生长得到促进,但水分严重减少对其生长和光合生理有抑制作用,但地上质量分数和水分利用效率却显著升高;(2)DN生境保水能力较好,随供水减少土壤水分含量下降较为平缓。叶片相对含水量、气孔导度和比叶面积在各水分处理之间差异不显著,但严重减水条件下总生物量、地上质量分数和水分利用效率均有回升。研究表明:浅而宽生境中植物倾向于通过提高地上部分的生长,保持较高的光合速率,并向地上部分分配较多生物量来应对水分胁迫;而深而窄生境中植物会充分利用土壤空间条件,促进根系的生长来提高对水分的吸收,从而维持植物的生长。     

Factors affecting photosynthesis and its seasonal variation in the seagrasses Cymodocea nodosa (Ucria) Aschers,and Posidonia oceanica (L.) Delile in the mediterranean

Measurements of photosynthesis, dark respiration, and leaf chlorophyll content were made in the laboratory on both shallow (1 to 5 m) and deep (25 to 33 m) leaves of Cymooceu nodosa (Ucria) Aschers, and Posidonia oceanica (L.) Delile in Malta in April and August. Light saturated photosynthetic rates in Cymodocea were similar in spring (18 μg C cm^?2h^?1) and summer (25μg Ccm^?2h^?1) if the 9 C increase in water temperature in summer is taken into account: however, photosynthetic rates in Posidonia were higher in spring than in summer, especially in shallow leaves which fixed ≈ 10 μg C cm^?2h^?1 in spring but less than half that in summer when rates of carbon accretion were close to compensation point. Levels of irradiance at which photosynthesis was light saturated (Iλ were ≈ 3 mW cm^?2 PAR for Cymodocea and 2 mW cm^?2 PAR for Posidonia: underwater irradiance at the lower depth limit for these plants (≈33 m) was ≈3 mW cm^?2 PAR. corresponding closely to the saturation irradiances. Compensation irradiance for both species was between 0.3 and 0.5 mW cm^?2 PAR.Photosynthesis in both species had a temperature optimum at about 30 C (slightly higher in Cymodocea in summer). Dark respiration rates were generally similar in spring and summer, in the region of 3 μg C cm^?2 h^?1 in Cymodocea and 1.5 to 2 μg C cm^?2 h^?1 in Posidonia. Increase in dark respiration rates with increased temperature was considerably greater in spring than in summer in both species. Photosynthesis was directly proportional to chlorophyll content in Posidonia in the range encountered (up to 58 μg Chl cm^?2) and the summer reduction in photosynthesis was closely correlated with reduction in chlorophyll content. It seems unlikely that environmental factors such as seasonal changes in light intensity, nutrient availability or water temperature were directly responsible for this loss of chlorophyll and it is suggested that this is a manifestation of general leaf senescence, probably induced by daylength changes but possibly enhanced by increased water temperature. Cymodocea showed a similar reduction in chlorophyll content in summer but this was not reflected in reduced photosynthesis. Thus, although Cymodocea may grow rapidly throughout the spring and summer with an overall productivity of 3.6 g C m^?2 day^?1 in shallow water, the luxuriant growths of Posidonia must develop in the first half of the year when a dense meadow may produce up to 2.1 g C m^?2 day^?1 in shallow water, declining to ?0.6 g C m^?2 day^?1 in summer.     

基于最小限制水分范围评价不同耕作方式对土壤有机碳的影响

以2009年吉林省德惠市中层黑土上进行了8a的田间定位试验小区土壤为研究对象,对免耕和秋翻两种耕作方式及玉米-大豆轮作和玉米连作两种种植方式下耕层有机碳进行分析,分别采用加权平均和分层两种方法计算最小限制水分范围(LLWR),用其评价不同耕作方式对土壤有机碳的影响.结果表明,免耕在玉米-大豆轮作和玉米连作下0-5 cm土壤有机碳含量分别比秋翻增加了15.2％和11.5％ (P＜0.05).采用加权平均法计算的LLWR值为0.148-0.166 cm3/cm3,不同耕作方式下玉米-大豆轮作的LLWR高于玉米连作且在两种种植方式下均表现出免耕小于秋翻的特点;利用分层法计算得到的LLWR值介于0.130-0.173 cm3/cm3之间,玉米-大豆轮作和玉米连作下免耕0-5 cm LLWR均显著小于秋翻,而5-30 cm LLWR数值免耕大于秋翻(P＞0.05);玉米-大豆轮作下0-30 cm各层LLWR均高于玉米连作.由于LLWR可以评价不同耕作方式对土壤有机碳的影响,因此采用加权平均法计算的LLWR可以客观的反映不同耕作处理尤其是种植方式对土壤有机碳的影响;而采用分层法计算的LLWR则更清晰的刻画了土壤表层与亚表层固碳能力的差异.     

Physiology of biofilms of thermophilic bacilli—potential consequences for cleaning

Thermophilic Bacillus species readily attached and grew on stainless steel surfaces, forming mature biofilms of >10^6.0 cells/cm² in 6 h on a surface inoculated with the bacteria. Clean stainless steel exposed only to pasteurized skim milk at 55 °C developed a mature biofilm of >10^6.0 cells/cm² within 18 h. When bacilli were inoculated onto the steel coupons, 18-h biofilms were 30 m thick. Biofilm growth followed a repeatable pattern, with a reduction in the numbers of bacteria on the surface occurring after 30 h, followed by a recovery. This reduction in numbers was associated with the production of a substance that inhibited the growth of the bacteria. Variations in the environment, including pH and molarity, affected the viability of the cells. Chemicals that attack the polysaccharide matrix of the biofilm were particularly effective in killing and removing cells from the biofilm, demonstrating the importance of polysaccharides in the persistence of these biofilms. Treatment of either the biofilm or a clean stainless steel surface with lysozyme killed biofilm cells and prevented the attachment of any bacteria exposed to the surface. This suggests that lysozyme may have potential as an alternative control method for biofilms of these bacteria.     

Development and evaluation of a diatom-conductivity model from lakes in West Greenland

1. The area around Kangerlussuaq (Søndre Strømfjord; West Greenland, 67°N 51°W) contains thousands of lakes ranging from coastal, dilute (conductivity < 30 μS cm^–1) oligotrophic systems to subsaline (～4000 μS cm^–1), closed basin lakes close to the ice sheet margin. In closed basins, salinity (or conductivity) is often a proxy for effective moisture, and thus palaeorecords of lake conductivity can provide valuable palaeoclimatic data. Little or nothing is known about the recent history of these lakes and hence it is difficult to evaluate how they will respond to effects of future changes. 2. Over 100 lakes have been sampled (1996–2000) between the ice sheet and the outer coast for a variety of water chemical and limnological variables. Surface sediments were taken from a subset of 40 lakes and analysed for diatoms. Diatom responses to 28 environmental variables were analysed by multivariate ordination techniques and indicate that the main gradient is highly correlated to conductivity (explaining ～12% of species variance). Despite the relatively short gradient (24–4072 μS cm^–1), diatom assemblages exhibit a clear response to conductivity. The most saline lakes do not contain a true saline flora. 3. We developed a range of weighted averaging (WA) and weighted‐averaging partial least squares (WA‐PLS) models from this training data set and found two component WA‐PLS models performed best. The effects of data transformation and omission of dissolution susceptible species (Diatoma spp.) on model performance were also examined. The error statistics for the preferred WA‐PLS (2) model (r²_jack=0.88, root mean square error of prediction (RMSEP)=0.217 log μS cm^–1) compare well with other published models. 4. A ²¹⁰Pb‐dated short core from a meromictic, subsaline lake (Braya Sø; location 67°N, 51°W, max. depth 23 m; conductivity 2600 μS cm^–1) was analysed for diatoms. Diatom preservation is poor and some taxa (e.g. Diatoma spp.) are badly corroded. Lake water conductivity was reconstructed using WA‐PLS models. Diatom‐inferred conductivity ranges from 1800 to 4400 μS cm^–1 over the last 600 years (extrapolated ²¹⁰Pb chronology). 5. The Kangerlussuaq area of West Greenland is an important area for palaeoclimatic research, located as it is between the Greenland ice sheet (ice core records) and the Davis Straits to the west. The development of a statistically robust transfer function for diatoms and conductivity will enable the reconstruction of conductivity from the many subsaline lakes around the head of the fjord and, hence, regional estimates of changing palaeoprecipitation.     

Studies on Tobacco Lignin

A study was made of such characteristics as infrared and ultraviolet absorption spectra, alkaline nitrobenzene oxidation, elementary composition and constituent groups of tobacco milled wood lignin (Japanese flue-cured and Matsukawa M. W. L.).

Methoxyl contents of mid-rib and stalk M. W. L. were 12.3~13.8% and 16.6~17.0% respectively. Infrared absorption spectra showed that the relation of the absorption intensities between at 1270 cm^?1 and at 1220 cm^?1 of stalk M. W. L. was 1270 cm^?1 < 1220 cm^?1 but, on the contrary, in the case of mid-rib M. W. L., the relation of the intensities was 1270 cm^?1 = 1220 cm^?1. Alkaline nitrobenzene oxidation indicated that the nuclear structures of tobacco M. W. L. were mainly composed of p-hydroxyphenyI and guaiacyl for mid-rib, and the former two and syringyl for stalk.     

Abundance, body composition and reproductive output of Gammarus minus (Crustacea: Amphipoda) in ten cold springs differing in pH and ionic content

1. A survey of thirty-two rheocrene springs in central Pennsylvania revealed that, Like Gammarus in lakes and streams, Gammarus minus is absent from springs with pH <6.0 and conductivity <25μS cm^?1 (total range in pH = 4.6–7.7, and in conductivity = 14–411 μS cm^?1). 2. In ten springs G. minus density was positively correlated with Ca²⁺ and Mg²⁺ hardness, but not with pH, unless three springs with either an exceptionally high velocity or extremely high densities of the potentially competing snail Fontigens nickliniana were omitted. 3. Adults were larger in the springs with few or no large predators than in those with more predators. In all ten springs, adult dry mass was unrelated to spring pH and ionic content, but brooding female dry mass covaried positively with Ca²⁺ and Mg²⁺ hardness in the five predator-poor springs. 4. Body water, Na and Ca contents and body mass/length ratios varied independently of spring pH and ionic content. Water content was inversely correlated with fat content, but even when expressed as a percentage of fat-free wet mass, it was unrelated to water chemistry. 5. In juveniles, males and non-brooding females, fat content varied independently of spring pH and ionic content, but in brooding females it was correlated with alkalinity and Ca²⁺ and Mg²⁺ hardness. The cost of reproduction in brooding females may have been a factor here; they had significantly lower per cent fat than did non-brooding females. Juvenile fat content did not differ significantly among spring populations, whereas adult fat content did. The per cent fat of brooding females covaried positively with body size among springs, and this was marginally true for non-brooding females, as well. The residuals of brooding female per cent fat against dry mass were not related to water chemistry. 6. Brood size (number of embryos in a brood) and brood mass varied significantly among populations, but independently of spring pH and ionic content. Both covaried positively with maternal body size among springs. The residuals of these relationships were unrelated to water chemistry, as was the percentage of females brooding. 7. G. minus from a pH 6 spring survived better and lost less body mass in acidic soft water than did those from a pH 7.6 spring. However, although G. minus has apparently been able to adapt (or acclimate) to pH 6 water it has failed to adapt to more acidic waters. A physiological or structural constraint may be involved because this species has probably had ample opportunity to evolve resistance to dilute acidic water. This hypothesis is consistent with the threshold effect observed: above pH 6 G. minus shows very little evidence of osmotic or metabolic stress, but beiow pH 6 viable populations apparently cannot survive at all. However, the gradual linear decrease in population density of G. minus with decreasing alkalinity and Ca²⁺ and Mg²⁺ hardness suggests that other factors may also be involved (e.g. a decrease in food quality).