Cold-Adapted Enzymes from Marine Antarctic Microorganisms

The Antarctic marine environment is characterized by challenging conditions for the survival of native microorganisms. Indeed, next to the temperature effect represented by the Arrhenius law, the viscosity of the medium, which is also significantly enhanced by low temperatures, contributes to slow down reaction rates. This review analyses the different challenges and focuses on a key element of life at low temperatures: cold-adapted enzymes. The molecular characteristics of these enzymes are discussed as well as the adaptation strategies which can be inferred from the comparison of their properties and three-dimensional structures with those of their mesophilic counterparts. As these enzymes display a high specific activity at low and moderate temperatures associated with a relatively high thermosensitivity, the interest in these properties is discussed with regard to their current and possible applications in biotechnology.     

Diversity of Polyketide Synthase Genes in the Genomes of Heterotrophic Microorganisms Isolated from Epilithic Biofilms of Lake Baikal

Many bacterial secondary metabolites, including pharmacologically promising compounds, are synthesized by polyketide synthases (PKS) enzyme complexes. In the present work, nucleotide sequences of the genes encoding 16S rRNA and PKS of heterotrophic bacterial strains isolated from epilithic biofilms of the littoral zone of Lake Baikal were determined. On the basis of molecular phylogenetic analysis of the 16S rRNA genes, six heterotrophic strains were identified: Serratia fonticola 1А and 10А, Pseudomonas umsongensis K10-2 and K10-3, Rheinheimera tilapiae K18, and Flavobacterium sp. 43-09. Sequencing of cloned amplification products for PKS gene cluster revealed 33 sequences. Genes involved in biosynthesis of antibiotics (difficidine, erythromycin, curacin, mixalamide, corallopyronin, and myxothiazol) and cytostatics (romidepsin, spiruchostatin, and disorazol) were found among homologous sequences. The low homology (50–83%) of the PKS amino acid sequences of Baikal bacteria with sequences from GenBank attests to the potential capability of strains to produce new, not yet studied bioactive compounds. The obtained results show that the studied strains may be of practical interest for biotechnological application.     

Free-Living Heterotrophic Nitrogen-Fixing Bacteria Isolated from Fuel-Contaminated Antarctic Soils

Five bacterial isolates enriched from fuel-contaminated Antarctic soils fixed nitrogen in the dark heterotrophically and nonsymbiotically. Two isolates utilized jet fuel vapors and volatile hydrocarbons for growth but not in N-deficient medium. Bacteria such as these may contribute to in situ biodegradation of hydrocarbons in Antarctic soils.     

Factors Affecting the Toxic Effect of Tin on Estuarine Microorganisms

Inorganic tin (SnCl₄·H₂O) is toxic to microbial populations obtained from estuarine sediments plated on nutrient medium solidified with either agar or purified agar. The use of gelatin as a gelling agent decreased the apparent toxicity of tin, and toxicity was markedly reduced in medium solidified with silica gel. There was no evidence that toxic agar-tin complexes were involved. Cd, Cu, Pb, Ni, and Zn exhibited similar toxicity patterns; therefore, toxicity levels determined in the laboratory should be extrapolated to the environment with caution. The addition of cysteine to the medium had no effect on tin toxicity. Serine or 3-hydroxyflavone enhanced toxicity, while humic acids or gelatin inhibited toxicity. Replacement of SO₄²⁻ with NO₃⁻ did not alter tin toxicity, but replacement of Cl⁻ with NO₃⁻ decreased tin toxicity. Thus, the toxic effect(s) of tin depend as much on the chemical speciation of the metal as on the total concentration of the metal in the medium.     

Some Factors Affecting Fermentation Capacity and Net Growth of Rumen Microorganisms

The fermentation rate of rumen microorganisms obtained from a sheep fed on hay was determined at different hours after feeding. The highest rate of fermentation was obtained immediately after feeding whereas the lowest rate was always before feeding. The effect of adding glucose, sucrose, and starch alone or in combination with urea, sodium citrate, or succinic acid on the maximal fermentation rate was studied. In no case was the maximal fermentation rate greater than that obtained in the presence of concentrate and straw. An investigation on the effect of C/N ratios, dilution, and substrate concentration on the net growth of rumen microorganisms was undertaken. The highest net growth value was obtained with a C/N ratio of 16.5 to 17.5, which is the ratio found for the ration fed to the animal. It was possible to increase net growth approximately threefold by using the proper dilution and fivefold by using the proper feed concentration.     

Factors Affecting Survival of Bacteriophage on Tomato Leaf Surfaces

The ability of bacteriophage to persist in the phyllosphere for extended periods is limited by many factors, including sunlight irradiation, especially in the UV zone, temperature, desiccation, and exposure to copper bactericides. The effects of these factors on persistence of phage and formulated phage (phage mixed with skim milk) were evaluated. In field studies, copper caused significant phage reduction if applied on the day of phage application but not if applied 4 or 7 days in advance. Sunlight UV was evaluated for detrimental effects on phage survival on tomato foliage in the field. Phage was applied in the early morning, midmorning, early afternoon, and late evening, while UVA plus UVB irradiation and phage populations were monitored. The intensity of UV irradiation positively correlated with phage population decline. The protective formulation reduced the UV effect. In order to demonstrate direct effects of UV, phage suspensions were exposed to UV irradiation and assayed for effectiveness against bacterial spot of tomato. UV significantly reduced phage ability to control bacterial spot. Ambient temperature had a pronounced effect on nonformulated phage but not on formulated phages. The effects of desiccation and fluorescent light illumination on phage were investigated. Desiccation caused a significant but only slight reduction in phage populations after 60 days, whereas fluorescent light eliminated phages within 2 weeks. The protective formulation eliminated the reduction caused by both of these factors. Phage persistence was dramatically affected by UV, while the other factors had less pronounced effects. Formulated phage reduced deleterious effects of the studied environmental factors.     

Factors Affecting Songbird Nest Survival in Northern Mixed-Grass Prairie

ABSTRACT Factors associated with the nest survival of mixed-grass prairie passerines are not well known, especially in the context of contemporary grassland management. We documented the nest survival of clay-colored sparrows (Spizella pallida), savannah sparrows (Passerculus sandwichensis), and bobolinks (Dolichonyx oryzivorus) in managed prairie in northwestern North Dakota, USA. We used logistic exposure models and an information-theoretic framework to estimate nest survival and evaluate support for mechanisms (grazing, temporal factors, nest parasitism, nest-site vegetation, and nest-patch factors) relevant to nest survival. Survival for the entire nesting interval (23–28 days) was low for clay-colored sparrow (18.2%), savannah sparrow (15.5%), and bobolink (3.5%). We found support for a cubic effect of nest age; survival of savannah and clay-colored sparrow nests was greatest during mid-incubation and least during the mid-nestling period. Parasitized clay-colored sparrow and bobolink nests had greater survival rates than nonparasitized nests. Nest survival of clay-colored sparrows increased with increasing vegetation height and density. For savannah sparrows, nest survival was lower when cattle were present than when cattle were absent. Characteristics of the nest patch did not have strong effects based on model coefficients and confidence intervals, though they appeared in many of the most supported models. Positive effects of vegetation height and density on nest survival of clay-colored sparrows and negative effects of cattle presence on nest survival of savannah sparrows suggest some detrimental effects of grazing. However, the need to restore and maintain intact prairies likely warrants the continuation of cattle grazing on conservation lands.     

Factors Affecting Nest Survival of Greater Sage-Grouse in Northcentral Montana

Abstract: We studied greater sage-grouse (Centrocercus urophasianus) in northcentral Montana, USA, to examine the relationship between nest success and habitat conditions, environmental variables, and female sage-grouse characteristics. During 2001-2003, we radiomarked 243 female greater sage-grouse, monitored 287 nests, and measured 426 vegetation plots at 4 sites in a 3,200-km² landscape. Nest survival varied with year, grass canopy cover, daily precipitation with a 1-day lag effect, and nesting attempt. In all years, daily survival rate increased on the day of a rain event and decreased the next day. There was temporal variation in nest success both within and among years: success of early (first 28 d of nesting season) nests ranged from 0.238 (SE = 0.080) in 2001 to 0.316 (SE = 0.055) in 2003, whereas survival of late (last 28 d of nesting season) nests ranged from 0.276 (SE = 0.090) in 2001 to 0.418 (SE = 0.055) in 2003. Renests experienced higher survival than first nests. Grass cover was the only important model term that could be managed, but direction and magnitude of the grass effect varied. Site, shrub and forb canopy cover, and Robel pole reading were less useful predictors of nest success; however, temporal and spatial variation in these habitat covariates was low during our study. We note a marked difference between both values and interpretations of apparent nest success, which have been used almost exclusively in the past, and maximum-likelihood estimates used in our study. Annual apparent nest success (0.46) was, on average, 53% higher than maximum-likelihood estimates that incorporate individual, environmental, and habitat covariates. The difference between estimates was variable (range = +8% to +91%). Management of habitats for nesting sage-grouse should focus on increasing grass cover to increase survival of first nests and contribute to favorable conditions for renesting, which should be less likely if survival of first nests increases.     

Bioextraction Dynamics of Potassium from Feldspar by Heterotrophic Microorganisms Isolated from Ceramic and Rhizospheric Soil

Bioleaching of potassium from feldspar was optimized for five fungal and two bacterial isolates for carbon, nitrogen, agitation, pH and incubation time. Fungal isolates SDS_R and SDS₃ and bacterial isolate SDS₁ released 34, 42 and 28 ppm K in sucrose-amended medium, respectively. Isolate SDS₃ took 10 days for the highest K solubilization, whereas SDS_R required 15 days under 150 rpm shaking condition. The pH of medium was found to decrease from 7.0 to 2.05 after the growth of the organisms. High-performance liquid chromatography analysis of the filtrate showed the presence of citric, tartaric and maleic acid, which could be responsible for the solubilization of potassium.     

Factors Affecting Survival of Clavibacter michiganensis subsp. sepedonicus in Water

The survival of Clavibacter michiganensis subsp. sepedonicus (Cms), the causal organism of bacterial ring rot in potato, was studied in water, to assess the risks for dissemination of Cms via surface water and infection of potato crops by irrigation. Cms was able to survive for a maximum period of 7 days in non‐sterile surface water at 10°C, a period during which Cms can be transported over long distances, but will also be strongly diluted. It is concluded that contamination of surface water with Cms can pose a threat on potato production only if aquatic host plants can multiply Cms in high densities. Survival of a fluidal and non‐mucoid strain was also studied in sterile ditch water and simulated ‘drainage water’, in sterile MilliQ water, in tap water, in physiological salt and in artificial xylem fluid. In addition, the influence of temperature and low oxygen conditions on persistence of Cms in some of these diluents was studied. A maximum survival period of 35 days was found for Cms in sterile tap water at 20°C, independent of the strain used. In the other diluents survival periods ranged between 0 and 21 days. Relatively poor survival was found in MilliQ water and artificial xylem fluid. Low temperatures of 4°C do not favour survival as it does in soil. Oxygen depletion affected survival detrimentally. Survival periods determined by agar dilution plating and a direct viable counting method, based on the use of indicators for esterase activity and membrane integrity were similar. Therefore, it was concluded that under the experimental conditions studied, Cms did not form cells in a viable but non‐culturable state.     

Vertical distribution in and isolation of bacteria from Lake Vanda: an Antarctic lake

Vertical distribution of bacteria in Lake Vanda, an Antarctic meromictic lake, was examined by the acridine orange epifluorescence direct count method. Total bacteria were 10⁴–10⁵ cells · ml^–1 in the water at 55 m depth and above, and increased drastically to 10⁷ cells · ml^–1 in the bottom water. Filamentous or long rodshaped bacteria occurred at a high frequency in the upper layers, but in the bottom layers most bacteria were coccoidal or short rods. Mean bacterial cell volume in water of between 10 m and 60 m deep was fairly large compared with common bacterial populations in seawater and lake water. Aerobic heterotrophic bacteria were recovered from the water of a depth of 30 m and above, and were assumed to belong to Caulobacter. Viable heterotrophic bacteria were not recovered from the high salinity deep water by media prepared with the same deep water. Phototrophic purple non-sulphur bacteria were isolated by enrichment cultures from water at 55 m depth.     

Factors Affecting Tropical Tree Damage and Survival after Catastrophic Wind Disturbance

The structure and dynamics of cyclone‐prone tropical forests are driven in part by variation in tree species resistance to and survival after wind‐induced structural damage. We determined the factors associated with tree damage and 3‐yr survival following Category 5 Cyclone Olaf on the Polynesian island of Ta'u, American Samoa. Despite sustaining a high rate of severe damage (34.6% of all trees snapped, 23.0% uprooted), system resilience was high with 74.3 percent stem survival overall and an annual mortality rate of 7.9 percent compared with 2.1 percent in nearby undisturbed late successional forest. Three‐yr survival rate of trees sustaining severe damage was 63.1 percent, compared to about 89 percent for trees sustaining only branch loss or defoliation. Three‐yr survival differed according to damage type, 78.5 percent after snapping vs. 38.4 percent after uprooting. Species differed widely in resistance to and survival after snapping and uprooting. Several species and individual traits were associated with the probability of snapping or uprooting; however, wood density was the only species trait consistently, and negatively, associated with the probability of sustaining either damage type. Survival after snapping was negatively associated with the proportion of the tree snapped off, which was determined by individual tree architecture. Species growth rate was negatively associated with survival after uprooting, indicating the importance of shade tolerance for survival after uprooting. Thus, whereas species traits seemed to exclusively underpin resistance to and survival after uprooting, they only partly accounted for snapping resistance, and did not determine the intensity of snap damage or survival after snapping. Our results highlight the importance of considering each damage type separately when considering ecological trade‐offs.     

Factors Affecting Golden-Cheeked Warbler Nest Survival in Urban and Rural Landscapes

ABSTRACT We evaluated hypotheses concerning temporal, landscape, and habitat effects on nest survival of golden-cheeked warblers (Dendroica chrysoparia) in an urban and a rural landscape during the breeding seasons of 2005 and 2006 in central Texas, USA. We found support for temporal effects of year and cubic effect of date and included them in candidate models that evaluated habitat and landscape effects. Nest survival was lower in 2006 than in 2005 and decreased nonlinearly as the breeding season progressed. We found support for edge effects with decreased nest survival nearer edges and in areas with increased open edge density (wooded habitat abutting open habitat) or decreased trail density. However, confidence intervals for the model-averaged odds ratios overlapped 1.0 for all edge variables. Overall daily survival rate was 0.964 (95% CI = 0.949-0.975), resulting in a 25-day period survival of 0.398 (95% CI = 0.269-0.524). Period survival in Austin's urban landscape (0.399, 95% CI = 0.270-0.526) was similar to survival in Fort Hood's rural landscape (0.396, 95% CI = 0.261-0.528). Both landscapes likely support self-sustaining populations based on reasonable assumptions for adult survival and number of nesting attempts. We suggest that some large urban preserves can provide breeding habitat of comparable quality to rural locations and recommend protecting large parcels (>100 ha) of breeding habitat with limited fragmentation and reducing the amount of wooded edge abutting open habitat to ensure nest survival regardless of their landscape context.     

Merriam's Turkey Nest Survival and Factors Affecting Nest Predation by Mammals

Abstract: Nest success is an important parameter affecting population fluctuations of wild turkeys (Meleagris gallopavo). Factors influencing mammalian predation on turkey nests are complicated and not well understood. Therefore, we assessed nest hazard risk by testing competing hypotheses of Merriam's turkey (M. g. merriami) nest survival in a ponderosa pine (Pinus ponderosa) ecosystem during 2001–2003. We collected nesting information on 83 female Merriam's turkeys; annual nest success averaged 50% for adult females (range = 45–59%) and 83% for yearling females (range = 75–100%). Proportional hazard modeling indicated that precipitation increased the hazard of nest mortality. However, estimated hazard of nest predation was lowered when incubating females had greater shrub cover and visual obstruction around nests. Coyotes (Canis latrans) were the primary predator on turkey nests. We hypothesize that precipitation is the best predictor of nest survival for first nests because coyotes use olfaction effectively to find nesting females during wet periods. Temporally, as the nesting season progressed, precipitation declined and vegetation cover increased and coyotes may have more difficulty detecting nests under these conditions later in the nesting period. The interaction of concealment cover with precipitation indicated that nest hazard risk from daily precipitation was reduced with greater shrub cover. Management activities that promote greater shrub cover may partially offset the negative effects of greater precipitation events.     

Distribution of Heterotrophic Bacteria in Lake Shira

A study of the horizontal and vertical distribution of heterotrophic bacteria in brackish Lake Shira in summer periods showed that mesophilic bacteria dominated in all areas of the lake, whereas psychrotolerant bacteria dominated in the metalimnion and hypolimnion of its central part. Nonhalophilic bacteria were mostly mesophilic and dominated in coastal waters. Most psychrotolerant bacteria were able to grow in the presence of 5–10% NaCl. Heterotrophic bacteria isolated in different regions of the lake were identified to a generic level. The isolates were classified into autochthonous and allochthonous microorganisms on the bases of their distribution pattern in the lake water, halotolerance, and ability to grow at low temperatures.     

Temporal and Vertical Difference in Factors Limiting Growth Rate of Heterotrophic Bacteria in Lake Biwa

Abstract Dilution bioassays were performed to examine the seasonal and vertical difference in the relative importance of factors limiting growth of heterotrophic bacteria in Lake Biwa. The lake water diluted by 0.2 μm lake filtrate (1:6.6) was enriched either with glucose (C), inorganic phosphorus (P), ammonium nitrogen (N), amino acids (AA), or a combination of these, and incubated for 2 days at the depths where lake water was collected (2.5, 20 and 30 m depths). Experiments showed that at 2.5 m, P was the most deficient resource for bacterial growth, but the magnitude of P limitation depended on water temperature. Among others, amino acids showed a slight but significant stimulation of bacterial growth rates during the fall. At 20 and 30 m, however, growth stimulation by resource addition was rarely detected. Vertically reciprocal translocation experiments revealed that the growth rate was limited by low temperature rather than resource supply at the greater depths. The results support a simple view that bacterial growth rate is basically regulated by water temperature, but high growth rate is not realized in summer because of resource depletion. The present study suggests that both temperature and P supply play a crucial role in biogeochemical cycling of organic matter in Lake Biwa through the bacterial growth rate. Received: 10 March 1999; Accepted: 14 May 1999