Rates of sulfate reduction and thiosulfate consumption in a marine microbial mat

Abstract The sulfur cycle in a microbial mat was studied by determining viable counts of sulfate-reducing bacteria, chemolithoautotrophic sulfur bacteria and anoxygenic phototrophic bacteria. All three functional groups of sulfur bacteria revealed a maximum population density in the uppermost 5 mm of the mat: 1.1 × 10⁸ cells of sulfate reducers cm⁻³ sediment, 2.0 × 10⁹ cells of chemolithoautotrophs cm⁻³ sediment, and 4.0 × 10⁷ cells of anoxygenic phototrophs cm⁻³ sediment. Bacterial dynamics were studied by sulfate reduction rate measurements, both under anoxic conditions (dark incubation) and oxic conditions (incubation in the light), and determination of the vertical distribution of the potential rate of thiosulfate consumption under oxic conditions. Sulfate reduction rates in the top 5 mm of the sediment were 566 nmol cm⁻³ d⁻¹ in the absence of oxygen, and 123 nmol cm⁻³ d⁻¹ in the presence of oxygen. In the latter case, the maximum rate was found in the 5–10-mm depth horizon (361 nmol cm⁻³ d⁻¹). Biological consumption of amended thiosulfate was rapid and decreased with depth, while in the presence of molybdate, thiosulfate consumption decreased to 10–30% of the original rate.     

Effectiveness of a steam-vacuum sanitizer for reducing Escherichia coli O157:H7 inoculated to beef carcass surface tissue

A steam-vacuum sanitizer reduced aerobic plate counts associated with bovine faecal contamination from 5.5 log₁₀ cfu cm⁻² to 3.0 ± 0.21 log₁₀ cfu cm⁻² on beef carcass short plates. The same beef carcass short plates inoculated wiht 7.6 ± 0.09 log₁₀ cfu cm⁻² Escherichia coli O157: H7 in faeces, yielded an average residual level of E. coli O157: H7 of 2.1 ± 0.21 log₁₀ cfu cm⁻² after steam-vacuum treatments. This study demonstrates the effectiveness of a steam-vacuum sanitizer for removing E. coli O157: H7 from beef carcasses.     

Soil oxygen flux measured polarographically in an Alaskan tussock tundra

Soil oxygen flux was measured polarographically in undisturbed tussock tundra and in an old vehicle track at Eagle Creek, Alaska. Because some polarograms did not show effective voltage plateaus, the polarographic method was tested for use in the tundra by experimental manipulation of soil water content and microbial oxygen demand. The response of soil oxygen flux was as predicted for artificially waterlogged systems, and the response to increased microbial demand indicated relatively high oxygen availability. Soil oxygen flux ranged from 25.9 ng O₂ cm⁻² min⁻¹ in Sphagnum mats to 3.9 ng O₂ cm⁻² min⁻¹ in unvegetated ruts of the vehicle track. Most sites in unvegetated ruts did not have measurable soil oxygen flux. In contrast, oxygen flux beneath Eriophorum vaginatum tussocks growing in the vehicle track averaged 10.7 ng O₂ cm⁻² min⁻¹.     

Short-term thermal acclimation induces adaptive changes in the inner mitochondrial membranes of fish skeletal muscle

In the oxidative muscles (musculi laterales superficiales) of crucian carp Carassius carassius acclimated for 6 weeks to either 5 or 25° C, the volume density and the surface density of fibres per tissue did not differ significantly between the control and experimental groups. The correlation ratio (μ²) for these values was below 50, 39·3 and 43·9 respectively. After acclimation to 5° C, the surface density of outer mitochondrial membrane per fibre increased significantly from 0·93 to 1·23m² cm⁻³ in the summer population but dropped from 0·94 to 0·67 m² cm⁻³ in the winter population. The surface density of outer mitochondrial membrane per mitochondrion increased from 3·24 to 4·52 m² cm⁻³ in summer fish. After acclimation to 25° C, the surface density of inner mitochondrial membranes per muscle fibre decreased from 4·04 to 1·79 m² cm⁻³ in summer fish and from 3·86 to 1·07 m² cm⁻³ in winter fish. The surface density of inner mitochondrial membranes per mitochondrion increased from 14·17 to 15·60 m²cm⁻³ in summer fish but dropped from 13·91 to 10·67 m² cm⁻³ in winter fish. Correlation matrices demonstrate a negative correlation of the surface density of outer mitochondrial membrane per mitochondrion with the volume density of mitochondria per fibre and temperature, suggesting cold-induced proliferation of small mitochondria. It was concluded that short-term cold acclimation increased surface area of the inner mitochondrial membranes in summer fish.     

Water vapour uptake by diapausing eggs of a tropical walking stick

Abstract. This study is the first to demonstrate the capacity of an arthropod egg, that of a tropical walking stick Extatosoma tiaratum (Macleuy), to absorb water vapour from the air. This species diapauses both as an early embryo and then again as a pharate first instar larva, and both stages are capable of absorbing water vapour. Water vapour absorption occurs at lower humidities and at a lower rate for an egg in early embryonic diapause (c. a_v 0.30, 0.516 mg h^-1δ_v^-1) than in the diapausing pharate first instar (c. a_v 0.60, 0.725 mgh^-1δa_v^-1) at 25C. In addition to having the capacity to gain water at very low vapour activities, water is efficiently conserved as indicated by the low rate of water loss (0.015% h^-1 in the early embryo and 0.046% h^-1 in the pharate larva at 25C). Eggs that have been killed lose water when held at a hydrating vapour activity, thus implying that active uptake contributes to net absorption. Wax block experiments suggest that water is absorbed over the entire chorionic surface. Eggs of five other insect species that were examined [Lymantria dispar (L.), Bombyx mori (L.), Antheraea polyphemus (Cram.), Oncopeltus fasciatus (Dallas) and Diaferomera femorata (Say)] lacked the ability to absorb atmospheric water.     

Responses of apple leaf stomata: a model for single leaves and a whole tree

Abstract. An empirical model of stomatal response to environmental factors was developed from measurements of stomatal conductance ( g _s) made in a leaf chamber under controlled conditions. Results presented in a companion paper (Warrit, Landsberg & Thorpe, 1980) indicated that the model could be written in terms of only two factors, photon flux density ( Q _p) and leaf to air vapour pressure gradient ( D ). The response of Q _p was hyperbolic and that to D linear; combining these the equation of the model is where g _r is a reference conductance, α is the slope of the response to D and β indicates the sensitivity of g _s response to Q _p. Values of α were 0.20 and 0.30 kPa⁻¹ in June and August; the corresponding values of β were 59 and 79 μmol m⁻² s⁻¹.
The model was tested against mean values of g _s obtained with a porometer in the field, using environmental measurements as inputs. Correspondence between measured and calculated values was good. Transpiration rates were calculated from the Penman-Monteith equation, with stomatal resistance values calculated from the model, and compared with gravimetric measurements of tree water use. It was shown that transpiration could be calculated with acceptable accuracy. The effects of variations in stomatal resistance on transpiration rates under a range of conditions were explored using the model and the Penman- Monteith equation.     

An integrated portable apparatus for the simultaneous field measurement of photosynthetic CO2 and water vapour exchange, light absorption and chlorophyll fluorescence emission of attached leaves

Abstract. A portable apparatus has been constructed to measure simultaneously the quantum yield of CO₂ assimilation, light absorption, chlorophyll fluorescence emission and water vapour exchange of attached intact leaves in the field. The core of the instrument is a light-integrating spherical leaf chamber which includes ports for a light source, photosynthetically active radiation sensor, fluorescence probes and gas inlet and outlet manifolds. Measurement of the quantum flux inside the empty chamber and with a leaf present allows determination of leaf absorptance. An open gas exchange system is employed using an infra-red analyser to measure leaf CO₂ exchange. Using a DC white light source the quantum yield of CO₂ assimilation based on absorbed light (φ_abs) may be determined rapidly in either ambient air or artificial gas mixtures. Inclusion of capacitance humidity probes into the gas inlet and outlet ports allows simultaneous determination of water vapour exchange and subsequent estimation of stomatal conductance to CO₂ and intercellular CO₂ concentration. Measurement of fluorescence emission by the sample leaf exposed to white light is achieved by a modulated fluorescence detection system. In addition to determination of the minimal, maximal and variable fluorescence levels, a further analysis allows the photochemical and non-photochemical components of fluorescence quenching, to be estimated. The theory and design of this apparatus is described in detail. The use of the apparatus in the field is demonstrated through a study of the photosynthetic performance of a maize and bean crop during the growing season and by analysis of the photosynthetic performance of crops subjected to nitrogen-stress and a herbicide treatment.     

Transmission of stridulatory signals of the burrower bugs, Scaptocoris castanea and Scaptocoris carvalhoi (Heteroptera: Cydnidae) through the soil and soybean

Abstract.  Males and females of the burrower bug species Scaptocoris castanea Perty and Scaptocoris carvalhoi Becker emit stridulatory signals when on the roots of soybean. The substrate-borne components of the signal can be recorded on the plant but not on the surrounding soil surface. The stridulatory apparatus is composed of the tergal plectrum (lima) and the stridulitrum (stridulatory vein) on the underside of the hind wings. The male plectrum has one ridge and the female lima has 13 ridges. Stridulitra of different species differ in the length and in the number of teeth. Rubbing of plectrum (lima) ridges over the stridulitrum in one or both directions produces pulse trains. The velocity of signals that are recorded less than 0.5 cm from the bug is below 0.013 mm s⁻¹ on the soil and below 0.066 mm s⁻¹ on the leaf surface. Broadband spectra have a dominant frequency of less than 1 kHz and subdominant peaks extending up to 7 kHz. The dominant frequency of the stridulatory signal transmitted through a plant decreases together with the proportion of its higher frequency spectral components. Signals are attenuated for 3–9 dB cm⁻¹ when transmitted through the soil or soybean leaf and for approximately 1 dB cm⁻¹ when transmitted through soybean stem.     

An experimental test of the eddy correlation technique over a Mediterranean macchia canopy

Abstract. Flux densities of water vapour and carbon dioxide were measured for a Mediterranean macchia canopy. Results show good agreement between the measured available energy and the sum of latent sensible and heat flux densities determined with the eddy correlation technique. Joint evaluation of the Bowen ratio, aerodynamic resistance, canopy resistance and the 'omega factor' suggests that the macchia canopy is intermediate in aerodynamic roughness between coniferous and deciduous canopies. Maximum daytime carbon flux densities ranged from -14 to -22(μnol m⁻² s⁻¹ on a ground area basis. The ratio of transpiration to assimilation (E/A) was a function of incident photo-synthetic photon flux density below about 400 μmol m⁻²s⁻¹ and above it was fairly constant at 272 mol mol⁻¹ (H₂O/CO₂). The relationship between carbon influx and canopy conductance was linear. Results show promising applications of the eddy correlation technique for evaluating physiological features of canopies, treated as unitary functional systems.     

Numerical abundance of myxomycetes (myxogastrids) in soils in the West of England

Abstract A most-probable-number technique was used to quantify the abundance of myxomycetes (myxogastrids) in soil samples taken from 23 'Sites of Special Scientific Interest' and one other site in the West of England. Associated organisms and soil conditions were also recorded. Sixteen of the 24 sites yielded myxomycete plasmodium-forming units (PFUs) in numbers averaging 230 cm⁻³ fresh soil. Where detectable in samples of grassland soil, the numbers ranged from 20–2750 cm⁻³, in woodland soil from 20–800 cm⁻³, and in sand dunes from 80–400 cm⁻³. Repeated sampling revealed changing numbers at single sites. The abundance of PFUs was correlated positively with numbers of soil amoebae, ciliates and nematodes, and with levels of magnesium and soil density, and negatively with organic matter content and levels of NH₄-nitrogen and phosphate. Each PFU probably corresponded to one or several uninucleate cells in the soil rather than to a plasmodium.     

Competition between anoxygenic phototrophic bacteria and colorless sulfur bacteria in a microbial mat

Abstract The populations of chemolithoautotrophic (colorless) sulfur bacteria and anoxygenic phototrophic bacteria were enumerated in a marine microbial mat. The highest population densities were found in the 0–5 mm layer of the mat: 2.0 × 10⁹ cells cm⁻³ sediment, and 4.0 × 10⁷ cells cm⁻³ sediment for the colorless sulfur bacteria and phototrophs, respectively. Kinetic parameters for thiosulfate-limited growth were assessed for Thiobacillus thioparus T5 and Thiocapsa roseopersicina M1, both isolated from microbial mats. For Thiobacillus T5, growing at a constant oxygen concentration of 43 μmol l⁻¹, μ_max was 0.336 h⁻¹ and K _s 0.8 μmol l⁻¹. Phototrophically grown Thiocapsa strain M1 displayed a μ_max of 0.080 h⁻¹ and a K _s of 8 μmol l⁻¹ when anoxically grown under thiosulfate limitation. In a competition experiment with thiosulfate as electron donor, Thiocapsa became dominant during a 10-h oxic/14-h anoxic regimen at continuous illumination, despite the higher affinity for thiosulfate of Thiobacillus .     

Generation of a reproducible nutrient-depleted biofilm of Escherichia coli and Burkholderia cepacia

An in vitro method of growing bacteria as a defined nutrient-depleted biofilm is proposed. The medium was defined nutritionally in terms of the quantitative composition and by the total amount of nutrient required to achieve a defined population size. Escherichia coli and Burkholderia cepacia were incubated on a filter support placed on a defined volume of solid medium. The change of biomass of the biofilm population was compared with the change in a planktonic culture. The size of the population in stationary phase was proportional to the concentration of limiting substrate up to 40 μmol cm⁻² glucose for E. coli and up to 2·7 × 10⁻⁹ mol cm⁻² iron for B. cepacia . Escherichia coli growing exponentially had a growth rate of μ = 0·30 h⁻¹ in a biofilm and μ = 0·96 h⁻¹ in planktonic culture. The growth rate, μ, for exponentially growing B. cepacia in a biofilm was 1·12 h⁻¹ and in planktonic culture 0·78 h⁻¹. This method allows the limitation of the size of a biofilm population to a chosen value.     

Low vapour pressure deficit reduces the beneficial effect of elevated CO2 on growth of N2-fixing alfalfa plants

Plant responses to elevated CO₂ can be modified by many environmental factors, but very little attention has been paid to the interaction between CO₂ and changes in vapour pressure deficit (VPD). Thirty-day-old alfalfa plants ( Medicago sativa L. cv. Aragón), which were inoculated with Sinorhizobium meliloti 102F78 strain, were grown for 1 month in controlled environment chambers at 25/15°C, 14 h photoperiod, and 600 µmol m⁻² s⁻¹ photosynthetic photon flux (PPF), using a factorial combination of CO₂ concentration (400 µmol mol⁻¹ or 700 µmol mol⁻¹) and vapour pressure deficit (0.48 kPa or 1.74 kPa, which corresponded to relative humidities of 85% and 45% at 25°C, respectively). Elevated CO₂ strongly stimulated plant growth under high VPD conditions, but this beneficial effect was not observed under low VPD. Under low VPD, elevated CO₂ also did not enhance plant photosynthesis, and plant water stress was greatest for plants grown at elevated CO₂ and low VPD. Moreover, plants grown under elevated CO₂ and low VPD had a lower leaf soluble protein and photosynthetic activity (photosynthetic rate and carboxylation efficiency) than plants grown under elevated CO₂ and high VPD. Elevated CO₂ significantly increased leaf adaxial and abaxial temperatures. Because the effects of elevated CO₂ were dependent on vapour pressure deficit, VPD needs to be controlled in experiments studying the effect of elevated CO₂ as well as considered in the extrapolations of results to a warmer, high-CO₂ world.     

Seasonal patterns of viruses, bacteria and dissolved organic carbon in a riverine wetland

SUMMARY 1. Viral and bacterial abundances were studied in relation to environmental attributes over an annual period, for both planktonic and attached (sediment, aquatic macrophyte and submerged wood) habitats, in a riverine wetland.
2. Annual mean abundance of planktonic viruses ranged from 2.3 × 10⁵−3.8 × 10⁵ particles mL⁻¹ and varied according to sampling site. Significant seasonal patterns in viral abundance were evident and appeared to be linked to variations in bacterial abundance, dissolved organic carbon and inorganic nutrients.
3. Annual mean abundance of viruses associated with surfaces ranged from 1.3 × 10⁶ particles cm⁻² on aquatic macrophytes to 1.1 × 10⁷ particles cm⁻² on wood and also showed seasonal patterns. The difference in viral dynamics among the different sites emphasizes the importance of considering habitat diversity within wetlands when examining microbial communities.     

Reduction of Brochothrix thermosphacta on beef surfaces following immobilization of nisin in calcium alginate gels

Lean and adipose beef carcass tissues inoculated with Brochothrix thermosphacta (BT) (approx. 4.50 log₁₀ cfu cm⁻²) were left untreated (U) or treated with 100 μg ml⁻¹ nisin (N), calcium alginate (A) or 100 μg ml⁻¹ nisin immobilized in a calcium alginate gel (AN). Tissue samples were refrigerated after treatments and bacterial populations and nisin activity were determined at 0, 1, 2 and 7 d. U, A and N treatments of lean and adipose tissues did not suppress bacterial growth (>6 log₁₀ cfu cm⁻² by day 7) while treatments of lean and adipose tissues with AN suppressed bacteria (>2.42 log₁₀ cfu cm⁻² by day 7). Bacteriocin titres from both tissues were higher in AN vs N samples after the 7 d incubation. This study demonstrates that immobilization of nisin in a gel may be a more effective delivery system of a bacteriocin to the carcass surface than direct application.     

Electroporation of abalone sperm enhances sperm-DNA association

The ability of sperm from the black-footed abalone Haliotis iris to take up foreign DNA in solution has been demonstrated. The efficiency of DNA uptake is related to the conditions of electroporation, including field strength (625 V cm⁻¹, 1000 V cm⁻¹), pulse length (18.6 ms, 27.4ms) and number of pulses (1, 2), and DNA concentration (20, 100 μg ml⁻¹). Sperm motility decreased with increased field strength and pulse number. At a field strength of 625 V cm⁻¹, neither the pulse length nor pulse number enhanced DNA uptake. A 40% enhancement in DNA uptake was observed when the sperm were shocked at 1000 V cm⁻¹ with two long pulses (27.4 ms each). Linear regression analysis revealed that pulse number ( p = 0.013) and field strength ( P =0.039) were the most important factors in sperm–DNA interaction. Higher DNA concentration enhanced sperm DNA uptake irrespective of field strength, pulse length and pulse number. The optimal electroporation conditions for DNA uptake were 1000 V cm', with two pulses of 27.4 ms each, and a DNA concentration of 100 μg ml⁻¹.     

Electrofusion of protoplasts from Aspergillus nidulans

Abstract Electrical parameters were determined and quantified for the stimulation of the optimum alignment and fusion of Aspergillus nidulans protoplasts. In a non-homogeneous alternating electrical field A. nidulans protoplasts aligned to form pearl chains associated with the electrodes of the fusion chamber. Most protoplasts were in pearl chains in an alignment field frequency of 3.0 MHz but maximum pair formation occurred at 1.0 MHz. At a field strength between 100 and 1000 V · cm⁻¹ pearl chain formation occurred with minimal protoplast rotation or lysis. The application of DC pulses resulted in protoplast fusion. Most fusion events were observed after two 500 V · cm⁻¹ DC pulses with a 0.5 s interpulse period. Using 1 × 10³ protoplasts · cm⁻³ in a 7 μm fusion chamber a maximum of 17.2 ± 2.0% fusion events were achieved.     

Densities and swimbladder development of juvenile American eels, Anguilla rostrata (Lesueur) as related to energetics of migration

Glass eels and elvers of the American eel were negatively buoyant. Those adapted to sea water were more dense (1.072 ± 0.001 g cm⁻³) than those adapted to fresh water (1.061 ± 0.001 g cm⁻³). Adaptation to fresh water increased relative body water content, but did not account for the observed decrease in total body density. Histological examination revealed the presence of a potentially functional swimbladder in the glass eels, although this hydrostatic organ did not becomegas-filled until after freshwater residency had occurred. Calculation of lift as used in the selective tidal transport mechanism suggests that hydrodynamic compensation for horizontal swimming during the estuarine phase of migration is energetically adaptive.     

Inactivation of bacteria and yeasts on agar surfaces with high power Nd: YAG laser light

G.D. WARD, I.A. WATSON, D.E.S. STEWART-TULL, A.C. WARDLAW AND C.R. CHATWIN. 1996. Near infrared light from a high-powered, 1064 nm, Neodymium : Yttrium Aluminium Garnet (Nd : YAG) laser killed a variety of Gram-positive and Gramnegative bacteria and two yeasts, lawned on nutrient agar plates. A beam (crosssectional area, 1.65 cm²) of laser light was delivered in 10 J, 8 ms pulses at 10 Hz, in a series of exposure times. For each microbial species, a dose/response curve was obtained of area of inactivation vs energy density (J cm⁻²). The energy density that gave an inactivation area (IA) equal to 50% of the beam area was designated the IA₅₀-value and was plotted together with its 95% confidence limits. Average IA₅₀-values were all within a threefold range and varied from 1768 J cm⁻² for Serratia marcescens to 4489 J cm⁻² for vegetative cells of Bacillus stearothermophilus. There were no systematic differences in sensitivity attributable to cell shape, size, pigmentation or Gram reaction. At the lowest energy densities where inactivation was achieved for the majority of organisms (around 2000 J cm⁻²), no effect was observed on the nutrient agar surface, but as the energy density was increased, a depression in the agar surface was formed, followed by localized melting of the agar.     

A survey of the Van Mijen fiord, Svalbard, as habitat for ringed seals, Phoca hispida

The ringed seal breeding habitat of the Van Mijen fiord, Svalbard, Norway, was studied from 30 March to 22 May 1986. A Siberian husky dog was used to detect ringed seal birth lairs and breathing holes. Fifteen percent of the total fiord areas was sampled. We estimated the densitites of birth lairs and breathing holes to be 0.04 km^-2 and 1.30 km^-2, respectively. The Van Mijen fiord was a poor ringed seal breeding habitat compared with breeding habitats investigated both in Canada and other parts of Svalbard. The main reason is probably shallow snow depth and lack of structures such as pressure ridges around which enough snow could accumulate for the ringed seal females to be able to dig out their lairs. The number of seals inhabiting the area during the breeding period 1986 was approximately 125 animals.