Cultivation of Lemna gibba under desert conditions. I: Twelve months of continuous cultivation in open ponds

Duckweed, Lemna gibba, was grown in 12 m² shallow ponds in the Negev desert, during 12 months of continuous cultivation, beginning April 1984. Average monthly growth rates varied with the season of the year. The lowest daily yield, 2·6±0·4 g dry weight m⁻² day⁻¹, was obtained during January. Highest daily yields, 7·9±2·6 g dry weight m⁻² day⁻¹ and 7·0±1·2 g dry weight m⁻² day⁻¹, were obtained during September and May. A 35% decline of the yield was seen during midsummer (July), 4·8±1·2 g dry weight m⁻² day⁻¹. The average rate for the year was 5·15±1·7 g dry weight m⁻² day⁻¹. The protein content of the plants ranged from 30 to 38% per unit dry weight.Growth performance is discussed in relation to the prevailing climatic conditions.     

Interactions between gnathiid isopods,cleaner fish and other fishes on Lizard Island,Great Barrier Reef

The rate of emergence of micropredatory gnathiid isopods from the benthos, the proportion of emerging gnathiids potentially eaten by Labroides dimidiatus, and the volume of blood that gnathiids potentially remove from fishes (using gnathiid gut volume) were determined. The abundance (mean ±s.e .) of emerging gnathiids was 41·7 ± 6·9 m^?2 day^?1 and 4552 ± 2632 reef^?1 day^?1 (reefs 91–125 m²). The abundance of emerging gnathiids per fish on the reef was 4·9 ± 0·8 day^?1; but excluding the rarely infested pomacentrid fishes, it was 20·9 ± 3·8 day^?1. The abundance of emerging gnathiids per patch reef was 66 ± 17% of the number of gnathiids that all adult L. dimidiatus per reef eat daily while engaged in cleaning behaviour. If all infesting gnathiids subsequently fed on fish blood, their total gut volume per reef area would be 17·4 ± 5·6 mm³ m^?2 day^?1; and per fish on the reefs, it would be 2·3 ± 0·5 mm^?3 fish^?1 day^?1 and 10·3 ± 3·1 mm³ fish^?1 day^?1 (excluding pomacentrids). The total gut volume of gnathiids infesting caged (137 mm standard length, L_S) and removed from wild (100–150 mm L_S) Hemigymnus melapterus by L. dimidiatus was 26·4 ± 24·6 mm³ day^?1 and 53·0 ± 9·6 mm³ day^?1, respectively. Using H. melapterus (137 mm L_S, 83 g) as a model, gnathiids had the potential to remove, 0·07, 0·32, 0·82 and 1·63% of the total blood volume per day of each fish, excluding pomacentrids, caged H. melapterus and wild H. melapterus, respectively. In contrast, emerging gnathiids had the potential of removing 155% of the total blood volume of Acanthochromis polyacanthus (10·7 mm L_S, 0·038 g) juveniles. That L. dimidiatus eat more gnathiids per reef daily than were sampled with emergence traps suggests that cleaner fishes are an important source of mortality for gnathiids. Although the proportion of the total blood volume of fishes potentially removed by blood‐feeding gnathiids on a daily basis appeared to be low for fishes weighing 83 g, the cumulative effects of repeated infections on the health of such fish remains unknown; attacks on small juvenile fishes, may result in possibly lethal levels of blood loss.     

Assessing the faecal source sensitivity and specificity of ruminant and human genetic microbial source tracking markers in the central Ethiopian highlands

This study tested genetic microbial source tracking (MST) methods for identifying ruminant- (BacR) and human-associated (HF183/BacR287, BacHum) bacterial faecal contaminants in Ethiopia in a newly created regional faecal sample bank (n = 173). BacR performed well, and its marker abundance was high (100% sensitivity (Sens), 95% specificity (Spec), median log₁₀ 8·1 marker equivalents (ME) g⁻¹ ruminant faeces). Human-associated markers tested were less abundant in individual human samples (median: log₁₀ 5·4 and 4·2 (ME + 1) g⁻¹) and were not continuously detected (81% Sens, 91% Spec for BacHum; 77% Sens, 91% Spec for HF183/BacR287). Furthermore, the pig-associated Pig2Bac assay was included and performed excellent (100% Sens, 100% Spec). To evaluate the presence of MST targets in the soil microbiome, representative soil samples were tested during a whole seasonal cycle (n = 60). Only BacR could be detected, but was limited to the dry season and to sites of higher anthropogenic influence (log₁₀ 3·0 to 4·9 (ME + 1) g⁻¹ soil). In conclusion, the large differences in marker abundances between target and non-target faecal samples (median distances between distributions ≥log₁₀ 3 to ≥log₁₀ 7) and their absence in pristine soil indicate that all tested assays are suitable candidates for diverse MST applications in the Ethiopian area.     

Feeding Rate of Brachionus plicatilis O. F. Müller on Two Types of Food Depending on Ambient Temperature and Salinity

Feeding rates of Brachionus plicatilis were studied for two types of food — algae Monochrysis lutheri and baker's yeast Saccharomyces cerevisae. The main regularities of changes in filtration rate and ration were studied in small culture volumes (1 ml) for adult amictic females depending on food concentration (1, 2, 4, 8 and 16 · 10⁶ cells · ml⁻¹), ambient temperature (16 and 26 °C), and salinity (5, 10, 15, 20, 25 and 30 ppt). B. plicatilis ration did not depend on the salinity, but was largely determined by temperature and food concentration. It was found that at 16 and 26 °C the dependence of the ingestion rate (ration) on food concentration differed greatly. A hypothesis was suggested to explain this phenomenon. A critical concentration of both types of food at which the increase in the rotifer ration ceased is 4 · 10⁶ cells · ml^−1. This is the minimum “background” food concentration for B. plicatilis mass cultivation. The average rations measured at the concentration of M. lutheri and S. cerevisae of 4 · 10⁶ cells · ml⁻¹ where 1.3 ± 0.1 and 4.8 ± 1.3 μg dry weight. · ind⁻¹ · day⁻¹ at 26 °C and 0.54 ± 0.1 and 1.9 μg d. w. · ind⁻¹ · day⁻¹ at 16 °C, respectively. The rations obtained in the laboratory were corrected for the conditions of rotifer commercial production in the open field in summer time. The correct values were 0.86 and 0.72 μg d. w. · ind⁻¹ · day⁻¹ for algae and yeast, respectively.     

The effects of dietary additives on faecal levels of Lactobacillus spp., coliforms,and Escherichia coli,and faecal prevalence of Salmonella spp. and Campylobacter spp. in US production nursery swine1

Aims: In the United States, carbadox and copper sulfate are growth promoters commonly used in combination in nursery swine diets. Our aim was to determine how selected dietary additives affect selected bacterial populations and pathogens in nursery swine, and compare to larch extract, which contains potential antibacterial activities. Methods and Results: Piglets were weaned and sorted into one of the four treatments: (i) basal diet without antimicrobials; (ii) basal diet with carbadox + copper sulfate; (iii) basal diet + 1000 ppm larch extract; or (iv) basal diet + 2000 ppm larch extract. Diets were fed for a 4‐week period after weaning. In both trials, the carbadox + copper sulfate group consumed more feed over the 4‐week period relative to the other three diet groups (P < 0·05), but did not gain significantly more weight. Faecal shedding of Salmonella spp. was not affected by dietary supplement in either trial, but faecal shedding of Campylobacter spp. was the lowest for the carbadox + copper sulfate diet. In faecal samples collected at the end of each trial, Lactobacillus spp. cell counts for the basal and larch extract diets were nearly 1·0 log₁₀ g^?1 faeces greater (P < 0·05) than the carbadox + copper sulfate group, whereas the coliforms and Escherichia coli were nearly 1·0 log₁₀ g^?1 faeces lower (P < 0·05). Conclusions: Compared to basal fed animals, supplementation with carbadox + copper sulfate significantly altered faecal E. coli, coliform bacteria and Lactobacillus spp. Larch extract has no benefit up to 0·2% of diet in regard to pathogen shedding, whereas carbadox + copper sulfate decreased faecal shedding of Campylobacter spp. Significance and Impact of the Study: Current swine management practices in the United States may be beneficial to managing Campylobacter spp. shedding in nursery swine, but also result in significant changes in the resident gastrointestinal microflora.     

Latitudinal and altitudinal growth patterns of brown trout Salmo trutta at different spatial scales

Spatial variation in growth of stream‐dwelling brown trout Salmo trutta was explored in 13 populations using a long‐term study (1993–2004) in the Bay of Biscay drainage, northern Spain. The high variability in fork length (L_F) of S. trutta in the study area was similar to the body‐size range found in the entire European distribution of the species. Mean L_F at age varied: 0+ years, 57·4–100·7 mm; 1+ years, 111·6–176·0 mm; 2+ years, 155·6–248·4 mm and 3+ years, 194·3–290·9 mm. Average L_F at age was higher in main courses and lower reaches compared with small tributaries and upper reaches. Annual specific growth rates (G_L) were: 0+ to 1+ years, 0·634–0·825 mm mm⁻¹ year⁻¹; 1+ to 2+ years, 0·243–0·342 mm mm⁻¹ year⁻¹; 2+ to 3+ years, 0·166–0·222 mm mm⁻¹ year⁻¹, showing a great homogeneity. Regression models showed that water temperature and altitude were the major determinants of L_F at age variability within the study area. A broader spatial analysis using available data from stream‐dwelling S. trutta populations throughout Europe indicated a negative relationship between latitude and L_F of individuals and a negative interaction between latitude and altitude. These findings support previous evidence of the pervasive role of water temperature on the L_F of this species. Altitude appeared as the overall factor that includes the local variation of other variables, such as water temperature or food availability. At a larger scale, latitude was the factor that encompassed these environmental gradients and explained the differences in L_F of S. trutta. In summary, L_F at age in stream‐dwelling S. trutta decreases with latitude in Europe, the converse of Bergmann's rule.     

Isolation of faecal coliform bacteria from the American alligator (Alligator mississippiensis)

Aims: To determine whether American alligators (Alligator mississippiensis) are an unrecognized poikilothermic source of faecal coliform and/or potential pathogenic bacteria in South Carolina’s coastal waters. Methods and Results: Bacteria from the cloaca of American alligators, as well as bacteria from surface water samples from their aquatic habitat, were isolated and identified. The predominant enteric bacteria identified from alligator samples using biochemical tests included Aeromonas hydrophila, Citrobacter braakii, Edwardsiella tarda, Escherichia coli, Enterobacter cloacae, Plesiomonas shigelloides and putative Salmonella, and these were similar to bacteria isolated from the surface waters in which the alligators inhabited. Based on most‐probable‐number enumeration estimates from captive alligator faeces, faecal coliform bacteria numbered 8·0 × 10⁹ g^?1 (wet weight) of alligator faecal material, a much higher concentration than many other documented endothermic animal sources. Conclusions: A prevalence of enteric bacteria, both faecal coliforms and potential pathogens, was observed in American alligators. The high faecal coliform bacterial density of alligator faeces may suggest that alligators are a potential source of bacterial contamination in South Carolina coastal waters. Significance and Impact of the Study: These findings help to increase our understanding of faecal coliform and potential pathogenic bacteria from poikilothermic reptilian sources, as there is the potential for these sources to raise bacterial water quality levels above regulatory thresholds.     

KINETICS OF GROWTH AND DEATH IN ANABAENA FLOS-AQUAE (CYANOBACTERIA) UNDER LIGHT LIMITATION AND SUPERSATURATION

The kinetics of population growth and death were investigated in Anabaena flos-aquae (Lyngb.) Bréb grown at light intensities ranging from limitation to photoinhibition (5 W·m⁻² to 160 W·m⁻²) in a nutrient-replete turbidostat. Steady-state growth rate (μ, or dilution rate, D) increased with light intensity from 0.44·day⁻¹ at a light intensity of 5 W·m⁻² to 0.99·day⁻¹ at 20 W·m⁻² and started to decrease above about 22 W·m⁻², reaching 0.56·day⁻¹ at 160 W·m⁻². The Haldane function of enzyme inhibition fit the growth data poorly, largely because of the unusually narrow range of saturation intensity. However, it produced a good fit (P < 0.001) for growth under photoinhibition. Anabaena flos-aquae died at different specific death rates (γ) below and above the saturation intensity. When calculated as the slope of a v_x⁻¹ and D⁻¹ plot, where v_x and D are cell viability (or live cell fraction) and dilution rate, respectively; γ was 0.047·day⁻¹ in the range of light limitation and 0.103·day⁻¹ under photoinhibition. Live vegetative cells and heterocysts, either in numbers or as a percentage of the total cells, showed a peak at the saturation intensity and decreased at lower and higher intensities. The ratio of live heterocysts to live vegetative cells increased with intensity when light was limiting but decreased when light was supersaturating. In cells growing at the same growth rate, the ratio was significantly lower under light inhibition than under subsaturation and the cell N:C ratio was also lower under inhibition. The steady-state rate of dissolved organic carbon (DOC) production increased with light intensity. However, its production as a percentage of the total C fixation was lowest at the optimum intensity and increased as the irradiance decreased or increased. The rate and percentage was significantly higher under photoinhibition than limitation in cells growing at the same growth rate. About 22% of the total fixed carbon was released as DOC at the highest light intensity. No correlation was found between the number of dead cells and DOC.     

In situ rates of nitrate uptake by giant kelp,Macrocystis Pyrifera (L.) C. Agardh: Tissue differences,environmental effects,and predictions of nitrogen-limited growth

NO⁻₃ uptake rates were measured in situ for seven types of tissue of adult sporophytes of Macrocystis pyrifera (L.) C. Agardh. Uptake by mature blades of canopy fronds followed saturation kinetics. At near-saturation concentrations, mean rates of uptake by different tissues ranged from 0.1 to 2.1 μg-at.· g wet wt⁻¹ · h⁻¹ (7–68 ng-at. · cm⁻²· h⁻¹). Different tissues incubated under similar environmental conditions took up NO⁻₃ at different rates, indicating physiological differences. Uptake rates showed no apparent relation to tissue age or nutritional history, but were influenced by certain environmental factors: uptake was 44–48% slower in dark than at ambient mid-day light levels, and dark uptake was 40% slower by blades incubated at 12m depth than by blades incubated at the surface. These physiological and environmental differences resulted in generally rapid uptake by tissues located at or just below the surface and slower uptake by tissues deeper in the water column.Daily NO⁻₃ uptake by M. pyrifera populations was predicted using a model based on in situ NO⁻₃ uptake rates. According to predictions of the model, NO⁻₃ concentrations of 1–2 μM throughout the water column are required to support kelp growth at the 4% · day⁻¹ (wet wt) rate typical of inshore populations. Vertical stratification, with high NO⁻₃ concentrations only at near-bottom depths, would result in severe nitrogen-limitation of growth. Seasonal changes in frond size distribution do not greatly affect nitrogen-limited growth rate. The model was also used to evaluate potential fertilizing methods and problems in management of offshore kelp farms.     

Algal biomass production,N uptake and N2 fixation in a synthetic medium

An experiment was conducted in the growth chamber to quantify the biomass production, N removal and N₂ fixation from a synthetic medium by Chlamydomonas reinhardtii and Anabaena flos-aquae. Nitrogen was supplied at a concentration of 100 mg liter⁻¹ of NO₃⁻−¹⁵N and NH₄⁺−¹⁵ (3·5 atom %), respectively. After 21 days Chlamydomonas reinhardtii removed an average of 83·8 and 78·7 mg N liter⁻¹ as NO₃⁻ and NH₄⁺, respectively. Averages of 0·89 and 0·71 g liter⁻¹ (first batch), 1·63 and 0·95 g liter⁻ (second batch) algal biomass were collected from NO₃⁻ and NH₄⁺ media, respectively. Uptake rates of 0·11 mg ¹⁵N g⁻¹ algae day⁻¹ from NO₃⁻ medium and 0·10 mg ¹⁵N g⁻¹ algae day⁻¹ from NH₄⁺ medium were calculated. Algal cells grown in NO₃⁻ and NH₄⁺ medium contained 71 and 65 g N kg⁻¹ (first batch), 39 and 58 g N kg⁻¹ (second batch), respectively. Anabaena flos-aquae produced averages of 0·58 and 0·46 g liter⁻¹ (first batch), 0·55 and 0·48 g liter⁻¹ (second batch) after 14 days of growth from NO₃⁻ and NH₄⁺ media, respectively. Blue-green algal biomass contained higher N (81–98 g kg⁻¹) than green algae. Isotope dilution method for determining N₂ fixation indicated that 55% and 77% of total N of blue-green algae grown in NO₃⁻ and NH₄⁺ media, respectively, was derived from the atmosphere.     

Identification and quantification of viable Bifidobacterium breve strain Yakult in human faeces by using strain-specific primers and propidium monoazide

Aims: To develop a quick and accurate PCR‐based method to evaluate viable Bifidobacterium breve strain Yakult (BbrY) in human faeces. Methods and Results: The number of BbrY in faeces was detected by using strain‐specific quantitative real‐time PCR (qPCR) derived from a randomly amplified polymorphic DNA analysis. And using propidium monoazide (PMA) treatment, which combined a DNA‐intercalating dye for covalently linking DNA in dead cells and photoactivation, only viable BbrY in the faeces highly and significantly correlated with the number of viable BbrY added to faecal samples within the range of 10⁵–10⁹ cells per g of faeces was enumerated. After 11 healthy subjects ingested 10·7 log CFU of BbrY daily for 10 days, 6·9 (±1·5) log CFU g^?1 [mean (±SD)] of BbrY was detected in faeces by using strain‐specific transgalactosylated oligosaccharide–carbenicillin (T‐CBPC) selective agar medium. Viable BbrY detected by qPCR with PMA treatment was 7·5 (±1·0) log cells per g and the total number (viable and dead) of BbrY detected by qPCR without PMA treatment was 8·1 (±0·8) log cells per g. Conclusions: Strain‐specific qPCR with PMA treatment evaluated viable BbrY in faeces quickly and accurately. Significance and Impact of the Study: Combination of strain‐specific qPCR and PMA treatment is useful for evaluating viable probiotics and its availability in humans.     

Size and growth rate differences of larval Baltic sprat Sprattus sprattus collected with bongo and MIK nets

The effect of sampling with bongo (0·6 m diameter frame with 500 µm mesh) and Methot Isaac Kidd (MIK) (2 m diameter frame with 2 mm mesh finished with 500 µm codend) nets on standard length (L_S) range and growth rate differences was tested for larval Sprattus sprattus (n = 906, L_S range: 7·0–34·5 mm) collected during four cruises in the summer months of 2006, 2007, 2009 and 2010 in the southern Baltic Sea. Although the minimum size of larvae collected with the bongo and MIK nets was similar in each cruise (from c. 7 to 9 mm), the MIK nets permitted collecting larger specimens (up to c. 34 mm) than the bongo nets did (up to c. 27 mm). The growth rates of larvae collected with the bongo and MIK nets (specimens of the same size range were compared for three cruises) were not statistically different (mean = 0·55 mm day^?1, n = 788, L_S range: 7·0–27·4 mm), which means the material collected with these two nets can be combined and growth rate results between them were compared.     

The influence of irradiance,photoperiod and temperature on the growth kinetics of three planktonic diatoms

The influence of irradiance, photoperiod and temperature was determined for the growth kinetics of the diatoms Aulacoseira subarctica, Stephanodiscus astraea and Stephanodiscus hantzschii and the results compared with those of cyanobacteria. Irradiance and photoperiod relationships were qualitatively similar to those for cyanobacteria in that: (1) growth rate (K) was proportionally greater under short photoperiods, with ratios of K under continuous light to K under 3:21 light:dark (LD) cycles of 1·50, 1·80 and 2·96 for A. subarctica, S. astraea and S. hantzschii respectively; (2) at subsaturating irradiances, K was proportional to irradiance and independent of temperature with a negligible predicted maintenance growth rate requirement. Apparent growth efficiencies (GE) at subsaturating irradiances were 0·26±0·03, 0·42±0·03 and 0·50±0·03 divisions mol^-1m² for A. subarctica, S. astraea and S. hantzschii with the values for Stephanodiscus species comparable to values for Oscillatoria species. Under a 3:21 LD cycle at 4 °C, light-saturated growth rates were 0·066±0·004, 0·197±0·033 and 0·285±0·018 divisions day^-1 for A. subarctica, S. astraea and S. hantzschii. S. hantzschii growth rate at 4 °C exceeded maximum Oscillatoria growth rates at 23 °C and the S. astraea growth rate at 4 °C was equivalent to O. agardhii growth rate at 20 °C. Temperature increases above 4 °C gave Q₁₀ values between 4 °C and 12 °C of 3·68, 2·39 and 1·92 for A. subarctica, S. astraea and S. hantzschii, but higher temperatures resulted in minor increases in K. S. astraea growth rate peaked at 16 °C, declining sharply at higher temperatures. February to March in situ growth rates in Lough Neagh, mean temperature 4·3 °C, showed that the A. subarctica in situ K of 0·058 divisions day^-1 was close to the laboratory K at 4 °C, but that S. astraea in situ K of 0·101 divisions day^-1 was lower than the laboratory K at 4 °C.     

Arthrospira platensis biomass with high protein content cultivated in continuous process using urea as nitrogen source

Aims: Arthrospira platensis has been studied for single‐cell protein production because of its biomass composition and its ability of growing in alternative media. This work evaluated the effects of different dilution rates (D) and urea concentrations (N₀) on A. platensis continuous culture, in terms of growth, kinetic parameters, biomass composition and nitrogen removal. Methods and results: Arthrospira platensis was continuously cultivated in a glass‐made vertical column photobioreactor agitated with Rushton turbines. There were used different dilution rates (0·04–0·44 day^?1) and urea concentrations (0·5 and 5 mmol l^?1). With N₀ = 5 mmol l^?1, the maximum steady‐state biomass concentration was1415 mg l^?1, achieved with D = 0·04 day^?1, but the highest protein content (71·9%) was obtained by applying D = 0·12 day^?1, attaining a protein productivity of 106·41 mg l^?1 day^?1. Nitrogen removal reached 99% on steady‐state conditions. Conclusions: The best results were achieved by applying N₀ = 5 mmol l^?1; however, urea led to inhibitory conditions at D ≥ 0·16 day^?1, inducing the system wash‐out. The agitation afforded satisfactory mixture and did not harm the trichomes structure. Significance and Impact of the Study: These results can enhance the basis for the continuous removal of nitrogenous wastewater pollutants using cyanobacteria, with an easily assembled photobioreactor.     

Lipid and biomass production by the halotolerant microalga Nannochloropsis salina

The effect of environmental factors on cell-lipid content, on the growth rate and on the overall productivity of Nannochloropsis salina was tested in the laboratory and in outdoor cultures. Under optimum conditions in the laboratory, the maximum growth rate (μ_max) was 0·030 h⁻¹, which corresponds to a doubling time of 23 h. Cellular lipid content was affected by the phase of growth and the temperature, but not by nitrogen starvation, pH or the source of sea water. The most important factor affecting the output rate of biomass was the cell concentration. The maximum biomass productivity obtained in outdoor ponds was 24·5 g·m⁻²·day⁻¹, and the lipid production rate was 4·0 g m⁻²·day⁻¹.     

Synthesis of hydroxyethyl starch 200/0.5-loaded albumin nanoparticles: biocompatibility and interaction mechanism

We aimed to synthesize hydroxyethyl starch (HES) 200/0.5-loaded bovine serum albumin nanoparticles (HBNs) and investigate the compatibility and binding mechanism in simulated physiological environments. Here, to elucidate the morphology, biocompatibility, and formation mechanism of HBNs, techniques such as scanning electron microscopy, haemolysis test, fluorescence, and circular dichroism spectroscopy were applied. The thermodynamic parameters at body temperature (ΔS° = −26.7 J·mol⁻¹·K⁻¹, ΔH° = −3.20 × 10⁴ J·mol⁻¹, and ΔG = −2.35 × 10⁴ J·mol⁻¹) showed a 1:1 binding stoichiometry, which was formed by hydrogen bonds and van der Waals interactions. In addition, the conformational analysis showed that the microenvironment of fluorophores was altered with the adaptational protein secondary structural changes. Energy transfer occurred from the fluorophores to HES with a high possibility. All these results provided accurate and complete primary data for demonstrating the interaction mechanisms of HES with BSA, which helps to understand its pharmaceutical effects in blood.     

Escherichia coli and enterococci are sensitive and reliable indicators for human,livestock and wildlife faecal pollution in alpine mountainous water resources

Aims: This study evaluated the applicability of standard faecal indicator bacteria (SFIB) for alpine mountainous water resources monitoring. Methods and Results: Escherichia coli, enterococci (ENTC) and Clostridium perfringens were investigated by standard or frequently applied phenotypic and genotypic methods in a broad range of animal and human faecal sources in a large alpine mountainous area. Clostridium perfringens occurred only in human, livestock and carnivorous source groups in relevant average concentrations (log 4·7–7·0 CFU g^?1) but not in herbivorous wildlife sources. Escherichia coli proved to be distributed in all faecal source groups with remarkably balanced average concentrations (log 7·0–8·4 CFU g^?1). Except for single faecal samples from the cattle source group, prevalence rates for ENTC source groups were generally >87% with average concentrations of log 5·3–7·7 CFU g^?1. To test the faecal indication capacity in the environment, faecal prevalence data were comparatively analysed with results from the concurrently performed multi‐parametric microbial source tracking effort on karst spring water quality from the investigated alpine mountainous catchment ( Reischer et al. 2008 ; Environ Microbiol 10:2598–2608). Conclusion: Escherichia coli and enterococci are reliable faecal indicators for alpine mountainous water resources monitoring, although E. coli is the more sensitive one. Clostridium perfringens did not prove to be an indicator of general faecal pollution but is suggested a conservative microbial source tracking marker for anthropogenic faecal influence. Significance and Impact of the Study: Applicability of SFIB is currently hotly debated. This is the first study providing comprehensive information on the applicability of SFIB at alpine mountainous habitats.     

Grazing behavior of rangeland beef cows differing in milk production

Differences in grazing behavior among Hereford (HH), 50% Angus-50% Hereford (AH), 50% Simmental-50% Hereford (1S1H) and 75% Simmental-25% Hereford (3S1H) lactating cows grazing foothill range were measured using vibracorders and pedometers to estimate daily grazing hours and distance traveled, respectively. There were four 10-day grazing periods during July and August. Cow weight, calf weight and milk production estimates were taken prior to Periods 1 and 4. During Periods 1, 2 and 4, the 48 cows were located on a daily basis between 06.00 and 09.00 h, and each animal location was recorded on gridded aerial photo maps. Pasture use was defined as the amount of total area utilized for grazing for the four periods.The overall mean for daily grazing hours was 9.4 h day⁻¹ (633 observations) and no significant breed type differences were found. Daily grazing hours increased from 8.0 to 10.0 h day⁻¹ during the 40-day observation period. Milk production and calf age significantly influenced daily grazing hours and the partial regression coefficients were 0.05 h·day⁻¹·kg·day⁻¹ and −0.02 h·day⁻¹·day⁻¹, respectively. The overall mean distance traveled was 4.7 km day⁻¹ (82 observations) and followed the same trend as daily grazing hours. For each kilometer of travel, 2h were spent grazing during the four grazing periods. The 1S1H cows traveled less (P < 0.05) than HH, AH and 3S1H cows. Breed type means were 5.0, 4.8, 4.1 and 4.8 km day⁻¹ for HH, AH, 1S1H and 3S1H cows, respectively. The overall mean for pasture use was 103 ha per cow (47 observations) and breed type was not significant in explaining variation in pasture use. For each kilogram increase in calf weight (adjusted for cow condition), pasture use increased by 0.5 ha (P < 0.05). Thus, the grazing behavior of different breed types under rangeland conditions was similar.     

The study of renin–angiotensin–aldosterone in experimental diabetes mellitus

It is generally accepted that hypertension and other vascular pathologies increase in diabetes mellitus (DM) patients as a result of the renin–angiotensin–aldosterone (RAA) system. In this study, changes in the renin‐angiotensin‐aldosterone (RAA) system level was determined in Streptozotocin (STZ)‐injected rats. A total of 46 female Wistar albino rats (180–220 g body weight) was utilized in these experiments. STZ was given intraperitoneally to induce diabetes in rats. Streptozotocin (60 mg kg⁻¹ body weight) was dissolved in 0·1 m citrate–‐phosphate buffer (pH 4–5). The non‐diabetic rats were injected with sterilized buffer alone to act as a control group. Blood glucose levels were 398±8·2 mg dl⁻¹, 488±11·75 mg dl⁻¹ and 658±29·6 mg dl⁻¹ at days 3, 12 and 30 respectively. The level of plasma renin activity (PRA) was measured as 7·69±1·07 ng ml⁻¹ h⁻¹; 1·82±0·22 ng ml⁻¹ h⁻¹ and 0·67±0·12 ng ml⁻¹ h⁻¹ at days 3, 12 and 30, respectively. These values showed that the PRA levels are decreased with increased time period. Serum angiotensin converting enzyme (ACE, E.C. 3.4.15.1) levels were increased at days 12 and 30 (p<0·05 and p<0·005), whereas serum aldosterone levels were increased at days 3 and 12 (p<0·05). The level of urea and creatinine increased at days 12 and 30 (p<0·05 and p<0·005, respectively) when compared to the control group. The data from these experiments indicate that the PRA level decreased whereas ACE activity level increased in diabetic rats compared with the control. Aldosterone levels increased at the first stage of the experiment, but then decreased by the end of the experiment as a result of changes in renin and ACE levels. Copyright © 1999 John Wiley & Sons, Ltd.     

Most probable number methodology for quantifying dilute concentrations and fluxes of Salmonella in surface waters

Aims: To better understand and manage the fate and transport of Salmonella in agricultural watersheds, we developed a culture‐based, five tube–four dilution most probable number (MPN) method for enumerating dilute densities of Salmonella in environmental waters. Methods and Results: The MPN method was a combination of a filtration technique for large sample volumes of environmental water, standard selective media for Salmonella and a TaqMan confirmation step. This method has determined the density of Salmonella in 20‐l samples of pond inflow and outflow streams as low as 0·1 MPN l^?1 and a low 95% confidence level 0·015 MPN l^?1. Salmonella densities ranged from not detectable to 0·55 MPN l^?1 for pond inflow samples and from not detectable to 3·4 MPN l^?1 for pond outflow samples. Salmonella densities of pond inflow samples were associated with densities of Escherichia coli and faecal enterococci that indicated stream contamination with faeces and with nondetectable pond outflow densities of the faecal indicator bacteria. The MPN methodology was extended to flux determinations by integrating with volumetric measurements of pond inflow (mean flux of 2·5 l s^?1) and outflow (mean flux of 5·6 l s^?1). Fluxes of Salmonella ranged from 100 to greater than 10⁴ MPN h^?1. Conclusions: This is a culture‐based method that can detect small numbers of Salmonella in environmental waters of watersheds containing animal husbandry and wildlife. Significance and Impact of the Study: Applying this method to environmental waters will improve our understanding of the transport and fate of Salmonella in agricultural watersheds, and can be the basis of valuable collections of environmental Salmonella.