Pad Culture Studies on Skin Surfaces

S ummary . A method is described for obtaining replicate velvet pad cultures of skin and other surfaces using malleable pads. This enables the bacterial flora of very irregular surfaces to be charted, and multiple replications on a range of selective media to be made. The application of the method to a number of clinical problems is reported.     

Protozoan pulses unveil their pivotal position within the soil food web

Protozoa are one of the most abundant groups of bacterivores within the soil and are responsible for mineralisation of bacterial biomass, having a large impact on C and N cycling. Little is known of their contribution to soil nutrient transfers or the identity of their consumers. Here, for the first time indigenous flagellates and ciliates, enriched to 83 atom% for (13)C and 10 atom% for (15)N, were introduced to soil cores from two different land managements, grassland and woodland with the same soil type, to trace the flow of protozoan C and N through the soil food web. Nematodes, Collembola, earthworms and insect larvae obtained the greatest amounts of C and N of protozoan origin, either through direct consumption or uptake of biomass post-cell death. Our results show that changes in management, affect the functioning of the soil food web and the utilisation of protozoa as a food source.     

A common soil flagellate (Cercomonas sp.) grows slowly when feeding on the bacterium Rhodococcus fascians in isolation, but does not discriminate against it in a mixed culture with Sphingopyxis witflariensis

Flagellates are very important predators on bacteria in soil. Because of their high growth rates, flagellate populations respond rapidly to changes in bacterial numbers. Previous results indicate that actinobacteria are generally less suitable than proteobacteria as food for flagellates. In this study, we investigated the growth of the flagellate Cercomonas sp. (ATCC 50334) on each of the two bacteria Sphingopyxis witflariensis (Alphaproteobacteria) and Rhodococcus fascians (actinobacteria) separately and in combination. The growth rate of the flagellate was lower and the lag phase was longer when fed with R. fascians than when fed with S. witflariensis. This supports our initial hypothesis that the actinobacterium is less suitable as food than the alphaproteobacterium. However, after longer periods of growth the peak abundance of flagellates was higher on R. fascians, indicating that the food quality of bacterial prey depends on the time perspective of the flagellate-bacterial interaction. There was no evidence that the flagellates selected against the actinobacterium when feeding in mixed cultures of the two bacteria. Experiments where flagellates were fed with washed bacterial cells or with bacteria growing with different substrate concentrations suggested that the low food quality of R. fascians is related both to the intrinsic cell properties and to the extracellular metabolites.     

Predation on Protozoa: its importance to zooplankton

Protozoa are an important component of both the nano- and microplanktonin marine and freshwater environments and are preyed upon byzooplankton, including suspension-feeding cope pods, some gelatinouszoopiankters and some first-feeding fish larvae. The clearancerates of suspension-feeding zooplankton for ciliates, in particular,are higher than for most phytoplankton. For at least some suspension-feedingzooplankton, protozoans are calculated to be quantitativelyan important component of the diet during certain seasons. Inlaboratory studies, protozoan components in the diet appearto enhance growth and survival of certain life-history stagesor enhance fecundity. These data suggest that protozoans arequalitatively as well as quantitatively important in the dietsof marine zooplankton. Most studies of predation on Protozoahave focused on the euphotic zone in nearshore waters. Predationon Protozoa is expected, however, to be particularly importantboth quantitatively and qualitatively in marine environmentsand seasons in which primary production is dominated by cells<5 µm in size, such as nearshore environments afterthe spring phytoplankton bloom, in oligotrophic waters, andin environments dominated by detritus-dominated food webs, suchas the deep sea. In detritus-dominated food webs, Protozoa maybe a source of essential nutrients and may thus facilitate utilizationof bacterial and detrital carbon by metazoan plankton.     

A method for turbidimetric measurement of bacterial growth in liquid cultures and agar plug diffusion cultures,using standard test tubes

Summary A method has been developed for turbidimetric measurement of bacterial growth in standard inexpensive test tubes with closures in-place. Liquid cultures and agar plug diffusion cultures can be assayed using an unmodified spectrophotometer. Growth curves of replicate cultures grown in test tubes, are reproducible with respect to similarity of curve shape, onset of logarithmic growth phase, and maximum growth.     

Nematophagous fungi from the maritime antarctic: factors affecting distribution

Soil collected from throughout the maritime Antarctic was analyzed to determine the effects of a range of abiotic and biotic soil parameters on the distribution of nematophagous fungi. Endoparasites were far more abundant than predatory fungi due to the greater efficiency of endoparasites in attracting and infecting nematodes, which resulted in a rapid completion of the infection cycle. This allowed endoparasites to colonise even the most exposed soil habitats where conditions favourable for nematode activity were restricted to only a few hours daily. 76% of all endoparasites isolated formed adhesive conidia. The success of this group of endoparasites was due to the ability of the conidia to attract nematodes.Spontaneous trap forming predators were far more abundant than nonspontaneous trap formers. The former have a clear competitive advantage over the latter, as they are able to attract nematodes to the conidia which subsequently germinate and form traps only when induced by direct chemical stimulation of nematodes. These predators are able to use internal reserves only and so rapidly respond to short periods when the soil is unfrozen and the nematodes are active.Although nematodes were utilized by all the species isolated, many were also capable of using Protozoa and Rotifera as a food source. While endoparasites were capable of infecting Rotifera, they were never observed infecting, or were isolated from soil containing, Protozoa. The distribution of endoparasites and spontaneous trap forming predators was largely independent of abiotic soil parameters. They were however significantly associated with the presence and abundance of suitable prey. These species survive solely on nematodes and therefore do not require prolonged periods of suitable soil conditions in order to produce vegetative mycelium to adsorb nutrients saprophytically from the soil, which is a significant ecological advantage over the other groups of microfungi found in the Antarctic.     

Biodiversity of Collembola and their functional role in the ecosystem

More than 6500 species of Collembola are known from throughout the world and these are only a small part of the still undescribed species. There are many checklists and catalogues of Collembola for smaller territories and entire continents. Biogeographical analyses have been made for some genera and smaller territories. The most serious problems for a global biogeographical analysis is the lack of enough records from immense territories of all continents. Local biodiversity of Collembola can be very high, reaching over 100 species in small mountain ranges. Sampling methods do not impede documenting biodiversity on a global scale. Collembola have well differentiated ecomorphological life-forms and feeding guilds which enable the functional role that Collembola play in ecosystems to be recognised in some degree. Collembola play an important role in plant litter decomposition processes and in forming soil microstructure. They are hosts of many parasitic Protozoa, Nematoda, Trematoda and pathogenic bacteria and in turn are attacked by different predators. They utilise as food Protozoa, Nematoda, Rotatoria, Enchytraeidae, invertebrate carrion, bacteria, fungi, algae, plant litter, live plant tissues, and some plant pathogens. Soil acidification, nitrogen supply, global climate change and intensive farming have greatly impacted collembolan diversity.     

Aphid effects on rhizosphere microorganisms and microfauna depend more on barley growth phase than on soil fertilization

This paper gives the first reports on aphid effects on rhizosphere organisms as influenced by soil nutrient status and plant development. Barley plants grown in pots fertilized with N but without P (N), with N and P (NP), or not fertilized (0) were sampled in the early growth phase (day 25), 1 week before and 1 week after spike emergence. Aphids were added 16 days before sampling was carried out. In a separate experiment belowground respiration was measured on N and NP fertilized plant–soil systems with aphid treatments comparable to the first experiment. Aphids reduced numbers of rhizosphere bacteria and fungal feeding nematodes 1 week before spike emergence. Before spike emergence, aphids reduced belowground respiration in NP treatments. These findings strongly indicate that aphids reduced allocation of photoassimilates to roots and deposition of root exudates in the growth phase of the plant. Contrary to this, 1 week after spike emergence numbers of bacteria, fungal feeding nematodes and Protozoa were higher in rhizospheres of plants subjected to aphids probably because aphids enhanced root mortality and root decomposition. Protozoa and bacterial feeding nematodes were stimulated at different experimental conditions with nematodes being the dominant bacterial grazers at N fertilization and Protozoa in the NP treatment before spike emergence.     

C and N mineralisation in the decomposer food webs of a European forest transect

Belowground processes are essential for the overall carbon and nitrogen fluxes in forests. Neither the functioning of the soil food web mediating these fluxes, nor its modulation by environmental factors is sufficiently understood. In this study the belowground carbon and nitrogen mineralisation of four European coniferous forest sites (northern Sweden to north‐east France) with different climate and N depositional inputs was analysed by investigating the soil food webs using field observations and modelling. The soil fauna directly contributed 7–13% to C mineralisation, among which the testate amoebae (Protozoa) made the largest contribution. Microbial grazing was suggested to have an important indirect effect by stimulating bacterial turnover. Due to relatively high C:N ratios of their substrate, bacteria immobilized N, while the fauna i.e. testate amoebae, nematodes, microarthropods and enchytraeids, counteracted this N immobilisation. Despite similar food web biomass, the sites differed with respect to food web structure and C and N flows. Model calculations suggested a significant influence of food web structure on soil ecosystem processes in addition to environmental factors and resource quality. Mineralisation rates were lowest at the low N input boreal site with a food web dominated by fungal pathways. Further south, as N availability increased, bacterial pathways became more important and the cycling of C and N was faster. The bioavailability of degradable C sources is suggested to be a limiting factor for microbial activity and overall mineralisation rates. In this respect, above‐ and belowground interactions e.g. transfers of labile C sources from the vegetation to the decomposer system deserve further attention. Our study revealed the combined effects of climate and nutrient inputs to ecosystems and the subsequent changes in the structure and functioning of the systems. If decomposition, and therefore carbon loss, is stimulated as a consequence of structural and/or nutritional changes, resulting for example from continuous industrial N emission, the storage capacity of forest ecosystems could be altered.     

Mobility of protozoa through narrow channels

Microbes in the environment are profoundly affected by chemical and physical heterogeneities occurring on a spatial scale of millimeters to micrometers. Physical refuges are critical for maintaining stable bacterial populations in the presence of high predation pressure by protozoa. The effects of microscale heterogeneity, however, are difficult to replicate and observe using conventional experimental techniques. The objective of this research was to investigate the effect of spatial constraints on the mobility of six species of marine protozoa. Microfluidic devices were created with small channels similar in size to pore spaces in soil or sediment systems. Individuals from each species of protozoa tested were able to rapidly discover and move within these channels. The time required for locating the channel entrance from the source well increased with protozoan size and decreased with channel height. Protozoa of every species were able to pass constrictions with dimensions equal to or smaller than the individual's unconstrained cross-sectional area. Channel geometry was also an important factor affecting protozoan mobility. Linear rates of motion for various species of protozoa varied by channel size. In relatively wide channels, typical rates of motion were 300 to 500 microm s(-1) (or about 1 m per hour). As the channel dimensions decreased, however, motilities slowed more than an order of magnitude to 20 microm s(-1). Protozoa were consistently observed to exhibit several strategies for successfully traversing channel reductions. The empirical results and qualitative observations resulting from this research help define the physical limitations on protozoan grazing, a critical process affecting microbes in the environment.     

Protozoa,Nematoda and Lumbricidae in the rhizosphere of Hordelymus europeaus (Poaceae): faunal interactions,response of microorganisms and effects on plant growth

Interactions among protozoa (mixed cultures of ciliates, flagellates and naked amoebae), bacteria-feeding nematodes (Pellioditis pellio Schneider) and the endogeic earthworm species Aporrectodea caliginosa (Savigny) were investigated in experimental chambers with soil from a beechwood (Fagus sylvatica L.) on limestone. Experimental chambers were planted with the grass Hordelymus europeaus L. (Poaceae) and three compartments separated by 45-m mesh were established: rhizosphere, intermediate and non-rhizosphere. The experiment lasted for 16 weeks and the following parameters were measured at the end of the experiment: shoot and root mass of H. europaeus, carbon and nitrogen content in shoots and roots, density of ciliates, amoebae, flagellates and nematodes, microbial biomass (SIR), basal respiration, streptomycin sensitive respiration, ammonium and nitrate contents, phosphate content of soil compartments. In addition, leaching of nutrients (nitrogen and phosphorus) and leachate pH were measured at regular intervals in leachate obtained from suction cups in the experimental chambers. Protozoa stimulated the recovery of nitrifying bacteria following defaunation (by chloroform fumigation) and increased nitrogen losses as nitrate in leachate. In contrast, protozoa and nematodes reduced leaching of phosphate, an effect ascribed to stimulation of microbial growth early in the experiment. Earthworms strongly increased the amount of extractable mineral nitrogen whereas it was strongly reduced by protozoa and nematodes. Both protozoa and nematodes reduced the stimulatory effect of earthworms on nitrogen mineralization. Microbial biomass, basal respiration, and numbers of protozoa and nematodes increased in the vicinity of the root. Protozoa generally caused a decrease in microbial biomass whereas nematodes and earthworms reduced microbial biomass only in the absence of protozoa. None of the animals studied significantly affected basal respiration, but specific respiration of microorganisms (O₂ consumption per unit biomass) was generally higher in animal treatments. The stimulatory effect of nematodes and earthworms, however, occurred only in the absence of protozoa. The sensitivity of respiration to streptomycin suggested that protozoa selectively grazed on bacterial biomass but the bacterial/fungal ratio appeared to be unaffected by grazing of P. pellio. Earthworms reduced root biomass of H. europaeus, although shoot biomass remained unaffected, and concentrations of nitrogen in shoots and particularly in roots were strongly increased by earthworms. Both nematodes and protozoa increased plant biomass, particularly that of roots. This increase in plant biomass was accompanied by a marked decrease in nitrogen concentrations in roots and to a lesser extent in shoots. Generally, the effects of protozoa on plant growth considerably exceeded those of nematodes. It is concluded that nematodes and protozoa stimulated plant growth by non-nutritional effects, whereas the effects of earthworms were caused by an increase in nutrient supply to H. europaeus.     

鄱阳湖地区生态系统服务权衡与协同关系的时空格局

鄱阳湖地区是中国最大淡水湖影响的区域,定量分析其生态系统服务权衡与协同关系对实现区域可持续发展具有重要意义.本研究首先基于遥感、土壤、气象、数字高程模型(DEM)等多源数据,定量测算2005—2015年鄱阳湖地区食物供给服务、土壤保持服务和产水服务,分析服务的时空格局及演变特征;然后运用相关分析、冷热点分析、空间制图方法探析1 km×1 km尺度上服务间权衡与协同关系的时空特征,并研究土地利用视角下生态系统服务及其相互关系的差异.结果表明: 研究期间,鄱阳湖地区食物供给服务增加,土壤保持服务减少,产水服务先增后减.空间分布上,食物供给服务在西北-东南方向上呈现“低-高-低-高”的分布格局,土壤保持服务呈现高低值集聚分布的空间格局,产水服务表现为南高北低的空间分布.鄱阳湖地区食物供给与土壤保持、食物供给与产水量是权衡关系,土壤保持与产水量是协同关系.食物供给与土壤保持呈现北部协同、南部权衡的空间格局,食物供给与产水量之间在空间上呈现出明显的权衡关系,土壤保持与产水量之间从北至南呈现协同-权衡-协同的“夹心式”分异规律.不同土地利用类型间,耕地与食物供给、园林地与土壤保持、水域与产水量之间均存在极显著协同关系,建设用地与3种服务皆是权衡关系.食物供给服务与土壤保持服务和产水服务之间在不同土地利用类型中皆以权衡为主,土壤保持与产水服务在耕地、园林地、草地及建设用地中以协同关系为主、在水域和未利用地中则以权衡关系为主.鄱阳湖地区多重生态系统服务热点区主要在东部和南部平原区,呈减少趋势.     

A Most Probable Number method (MPN) for the estimation of cell numbers of heterotrophic nitrifying bacteria in soil

A Most Probable Number (MPN) method was developed allowing for the first time estimation of populations of bacteria capable of heterotrophic nitrification. The method was applied to an acidic soil of a coniferous forest exhibiting nitrate production. In this soil nitrate production was unlikely to be catalyzed by autotrophic nitrifiers, since autotrophic ammonia oxidizers never could be detected, and autotrophic nitrite oxidizers were usually not found in appreciable cell numbers. The developed MPN method is based on the demonstration of the presence/absence of nitrite/nitrate produced by heterotrophic nitrifying bacteria during growth in a complex medium (peptone-meat-extract softagar medium) containing low concentrations of agar (0.1%). Both the supply of the growing cultures in MPN test tubes with sufficient oxygen and the presence of low agar concentrations in the medium were found to be favourable for sustainable nitrite/nitrate production. The results demonstrate that in the acidic forest soil the microbial population capable of heterotrophic nitrifcation represents a significant part of the total aerobic heterotrophic population. By applying the developed MPN method, several bacterial strains of different genera not previously described to perform heterotrophic nitrification have been isolated from the soil and have been identified by bacterio-diagnostic tests.     

Distribution, population dynamics and habitat use of the lesser pouched rat, Beamys hindei

The lesser pouched rat, Beamys hindei Thomas 1909, is one of Africa's rarest and least known rodents, recorded only from a few localities in Kenya and Tanzania. The results of this study show that B. hindei is more widely distributed than previously thought and occurs at high densities in suitable habitat. It breeds throughout the year, but maintains relatively constant population densities as recruitment rates are low. On account of its ability to cache food, it is well adapted to seasonally dry forests where food is in short supply for part of the year. The need for suitable soil in which to construct its burrows and dense vegetation cover may partly account for its patchy distribution. Morphological data collected during this study provide no evidence for separating B. hindei from B. major and suggest that the differences previously recorded between the two forms may be actually due to clinal variation in size from north to south.     

Microbial film development in a trickling filter

The transmission and scanning electron microscopes were employed to visualize the sequence of the biofilm development in the trickling wastewater filter. After the deposit of a small amount of debris upon a hard surface, the bacterial cells attach and develop the matrix on which the biofilm is formed. Protozoa invade the basic layer where they feed on the bacteria. The algae are seeded upon the bacterial matrix and grow so profusely that the bacteria must develop aerial colonies in the competition for food and oxygen. Destruction of the bacteria in the matrix and the weight and hydraulic pressure cause detachment of the biofilm and a new matrix must be developed.     

An improved method for assessing soil-fungus population density

Summary A rapid method for determining soil fungal populations is described. A 25-g equivalent of oven-dry soil, in 225 ml of water in a 500-ml aspirator bottle, is agitated on a magnetic stirrer and samples withdrawn with a syringe while the suspension is in motion. Individual drops are placed in petri dishes and mixed with a suitable culture medium. The average weight of soil per drop is determined and colony counts related to dry weight of soil as usual. This syringe method is simple, eliminates much of the error common in serial dilution and pipetting, and allows for a high degree of reproducibility and possible standardization.     

Mobility of Protozoa through Narrow Channels

Microbes in the environment are profoundly affected by chemical and physical heterogeneities occurring on a spatial scale of millimeters to micrometers. Physical refuges are critical for maintaining stable bacterial populations in the presence of high predation pressure by protozoa. The effects of microscale heterogeneity, however, are difficult to replicate and observe using conventional experimental techniques. The objective of this research was to investigate the effect of spatial constraints on the mobility of six species of marine protozoa. Microfluidic devices were created with small channels similar in size to pore spaces in soil or sediment systems. Individuals from each species of protozoa tested were able to rapidly discover and move within these channels. The time required for locating the channel entrance from the source well increased with protozoan size and decreased with channel height. Protozoa of every species were able to pass constrictions with dimensions equal to or smaller than the individual's unconstrained cross-sectional area. Channel geometry was also an important factor affecting protozoan mobility. Linear rates of motion for various species of protozoa varied by channel size. In relatively wide channels, typical rates of motion were 300 to 500 μm s⁻¹ (or about 1 m per hour). As the channel dimensions decreased, however, motilities slowed more than an order of magnitude to 20 μm s⁻¹. Protozoa were consistently observed to exhibit several strategies for successfully traversing channel reductions. The empirical results and qualitative observations resulting from this research help define the physical limitations on protozoan grazing, a critical process affecting microbes in the environment.     

Transparent Soil for Imaging the Rhizosphere

Understanding of soil processes is essential for addressing the global issues of food security, disease transmission and climate change. However, techniques for observing soil biology are lacking. We present a heterogeneous, porous, transparent substrate for in situ 3D imaging of living plants and root-associated microorganisms using particles of the transparent polymer, Nafion, and a solution with matching optical properties. Minerals and fluorescent dyes were adsorbed onto the Nafion particles for nutrient supply and imaging of pore size and geometry. Plant growth in transparent soil was similar to that in soil. We imaged colonization of lettuce roots by the human bacterial pathogen Escherichia coli O157:H7 showing micro-colony development. Micro-colonies may contribute to bacterial survival in soil. Transparent soil has applications in root biology, crop genetics and soil microbiology.     

Utilization of 4 Sugars Known to Occur in Activated Sludge by Aspidisca cicada (Ciliata, Hypotrichida); Effect on Division Rate

SYNOPSIS. Indiviluals of the hypotrich Aspidisca cicada (Müller) were separated from activated sludge and grown clonally. Lactose and sucrose increased its division rate; D-glucose and arabinose were inhibitory. All 4 sugars stimulated growth of the bacteria associated with A. cicada. Spectrophotometric detection of acid production showed that A. cicada utilized lactose and sucrose markedly, glucose only slightly, and arabinose not at all. The increased fission rate in the presence of lactose and sucrose is probably caused directly by these sugars providing a carbon source, and indirectly by increasing the supply of food bacteria. Retardation of growth of A. cicada in the presence of glucose or arabinose perhaps results from these sugars somehow making the food bacteria unavailable to A. cicada. Sugars found in activated sludge may inhibit or stimulate growth in populations of A. cicada , frequently one of the dominant Protozoa in activated sludge.