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Temperature control for kinetic refolding of heat-denatured ovalbumin.

The folding of heat-denatured ovalbumin, a non-inhibitory serpin with a molecular size of 45 kDa, was examined. Ovalbumin was heat-denatured at 80 degrees C under nonreducing conditions at pH 7.5 and then cooled either slowly or rapidly. Slow cooling allowed the heat-denatured ovalbumin to refold to its native structure with subsequent resistance to digestion by trypsin. Upon rapid cooling, by contrast, the heat-denatured molecules assumed the metastable non-native conformations that were susceptible to trypsin. The non-native species were marginally stable for several days at a low temperature, but the molecules were transformed slowly into the native conformation. Considering data from size-exclusion chromatography and from analyses of CD, intrinsic tryptophan fluorescence, and adsorption of the dye 1-anilinonaphthalene-8-sulfonate, we postulated that the non-native species that accumulated upon rapid cooling were compact but structureless globules with disordered side chains collectively as a folding intermediate. Temperature-jumped CD experiments revealed biphasic kinetics for the refolding process of heat-denatured ovalbumin, with the features of increasing and subsequently decreasing amplitude of the rapid and the slow phases, respectively, with the decrease in folding temperature. The temperature dependence of the refolding kinetics indicated that the yield of renaturation was maximal at about 55 degrees C. These findings suggested the kinetic partitioning of heat-denatured ovalbumin between alternative fates, slow renaturation to the native state and rapid collapse to the metastable intermediate state. Analysis of disulfide pairing revealed the formation of a scrambled form with non-native disulfide interactions in both the heat-denatured state and the intermediate state that accumulated upon rapid cooling, suggesting that non-native disulfide pairing is responsible for the kinetic barriers that retard the correct folding of ovalbumin.

Animals↗

Invasive species and coal bed methane development in the Powder River Basin, Wyoming.

One of the fastest growing areas of natural gas production is coal bed methane (CBM) due to the large monetary returns and increased demand for energy from consumers. The Powder River Basin, Wyoming is one of the most rapidly expanding areas of CBM development with projections of the establishment of up to 50,000 wells. CBM disturbances may make the native ecosystem more susceptible to invasion by non-native species, but there are few studies that have been conducted on the environmental impacts of this type of resource extraction. To evaluate the potential effects of CBM development on native plant species distribution and patterns of non-native plant invasion, 36 modified Forest Inventory and Analysis plots (each comprised of four 168-m2 subplots) were established in the Powder River Basin, Wyoming. There were 73 168-m2 subplots on control sites; 42 subplots on secondary disturbances; 14 on major surface disturbances; eight on well pads; and seven on sites downslope of CBM wells water discharge points. Native plant species cover ranged from 39.5 +/- 2.7% (mean +/- 1 SE) in the secondary disturbance subplots to 17.7 +/- 7.5% in the pad subplots. Non-native plant species cover ranged from 31.0 +/- 8.4% in the discharge areas to 14.7 +/- 8.9% in the pad subplots. The control subplots had significantly less non-native species richness than the combined disturbance types. The combined disturbance subplots had significantly greater soil salinity than the control sites. These results suggest that CBM development and associated disturbances may facilitate the establishment of non-native plants. Future research and management decisions should consider the accumulative landscape-scale effects of CBM development on preserving native plant diversity.

Coal Mining↗

Laboratory evaluation of the predation efficacy of native Australian fish on Culex annulirostris (Diptera: Culicidae).

The introduction and establishment of fish populations can provide long-term, cost-effective mosquito control in habitats such as constructed wetlands and ornamental lakes. The predation efficacy of 7 native Brisbane freshwater fish on 1st and 4th instars of the freshwater arbovirus vector Culex annulirostris was evaluated in a series of 24-h laboratory trials. The trials were conducted in 30-liter plastic carboys at 25+/-1 degrees C under a light:dark cycle of 14:10 h. The predation efficacy of native crimson-spotted rainbowfish Melanotaenia duboulayi (Melanotaeniidae), Australian smelt Retropinna semoni (Retropinnadae), Pacific blue-eye Pseudomugil signifer (Atherinidae), fly-specked hardyhead Craterocephalus stercusmuscarum (Atherinidae), firetail gudgeon Hypseleotris galii (Eleotridae), empire gudgeon Hypseleotris compressa (Eleotridae), and estuary perchlet Ambassis marianus (Ambassidae) was compared with the exotic eastern mosquitofish Gambusia holbrooki (Poeciliidae). This environmentally damaging exotic has been disseminated worldwide and has been declared noxious in Queensland. Melanotaenia duboulayi was found to consume the greatest numbers of both 1st and 4th instars of Cx. annulirostris. The predation efficacy of the remaining Australian native species was comparable with that of the exotic G. holbrooki. With the exception of A. marianus, the maximum predation rates of these native species were not statistically different whether tested individually or in a school of 6. Based on these data, M. duboulayi, H. compressa, and A. marianus warrant further investigation as biological control agents in pilot field trials.

Animals↗

Are invasive species the drivers of ecological change?

Invasive species are widely accepted as one of the leading direct causes of biodiversity loss. However, much of the evidence for this contention is based on simple correlations between exotic dominance and native species decline in degraded systems. Although appealing, direct causality is not the only possible interpretation. A plausible alternative hypothesis is that exotic dominance could be the indirect consequence of habitat modification driving native species loss. In a new paper, MacDougall and Turkington now provide the first direct test of whether invasive species are the drivers of community change, or merely 'passengers' along for the environmental ride.

Journal Article↗

Helminths of two native frog species (Rana chiricahuensis, Rana yavapaiensis) and one introduced frog species (Rana catesbeiana) (Ranidae) from Arizona.

The gastrointestinal tracts, lungs, urinary bladders, and body cavities of Rana catesbeiana (n = 25), Rana chiricahuensis (n = 25), and Rana yavapaiensis (N = 37) from Arizona were examined for helminths. Helminths representing 9 species of trematodes: Cephalogonimus brevicirrus, Glypthelmins quieta, Gorgoderina attenuata, Haematoloechus complexus, Haematoloechus langiplexus, Megalodiscus temperatus, Alaria sp., Clinostomum sp., and an unidentified strigeid; and 4 species of nematodes: Falcaustra catesbeianae, Rhabdias ranae, Physaloptera sp., and an unidentified ascarid were found. The helminth fauna of introduced R. catesbeiana differed markedly from that of native ranids. Helminths of R. chiricahuensis and R. yavapaiensis represent new host records. Arizona is a new locality record for C. brevicirrus, G. attenuata, H. complexus, H. longiplexus, M. temperatus, and R. ranae.

Animals↗

Covariation in leaf and root traits for native and non-native grasses along an altitudinal gradient in New Zealand.

Across 30 grassland sites in New Zealand that ranged from native alpine grasslands to low elevation improved pastures, there were consistent patterns of leaf and root traits and significant differences between native and non-native grasses. Plants of high altitude sites have low N concentrations in both their leaves and roots, have thick leaves and roots, yet no differences in tissue density or photosynthetic water use efficiency when compared to plants of low altitude sites. Both the leaves and roots of the low altitude plants were enriched in (15)N relative to the plants of higher altitude, indicating that the low-N set of traits is associated with a more closed N cycle at high altitude. A second independent set of correlations shows that plants of wetter habitats have lower photosynthetic water use efficiency (more negative partial differential (13)C) and lower leaf and root tissue density than the plants of drier sites. For both leaves and roots, plants of native species consistently had traits associated with lower resource availability: lower N concentrations, denser tissues, more negative partial differential (15)N, and more positive partial differential (13)C than non-native species. If root %N is correlated with root longevity as has been shown in other systems, root longevity may be able to be predicted from simple measurements of leaf %N, though a hysteresis in the relationship between leaf and root N concentrations may make prediction of high longevity roots difficult.

Adaptation, Physiological↗

Invasion, competitive dominance, and resource use by exotic and native California grassland species.

The dynamics of invasive species may depend on their abilities to compete for resources and exploit disturbances relative to the abilities of native species. We test this hypothesis and explore its implications for the restoration of native ecosystems in one of the most dramatic ecological invasions worldwide, the replacement of native perennial grasses by exotic annual grasses and forbs in 9.2 million hectares of California grasslands. The long-term persistence of these exotic annuals has been thought to imply that the exotics are superior competitors. However, seed-addition experiments in a southern California grassland revealed that native perennial species, which had lower requirements for deep soil water, soil nitrate, and light, were strong competitors, and they markedly depressed the abundance and fecundity of exotic annuals after overcoming recruitment limitations. Native species reinvaded exotic grasslands across experimentally imposed nitrogen, water, and disturbance gradients. Thus, exotic annuals are not superior competitors but rather may dominate because of prior disturbance and the low dispersal abilities and extreme current rarity of native perennials. If our results prove to be general, it may be feasible to restore native California grassland flora to at least parts of its former range.

California↗

Ion trap collisional activation of disulfide linkage intact and reduced multiply protonated polypeptides.

The presence of disulfide linkages in multiply charged polypeptide ions tends to inhibit the formation of structurally informative product ions under conventional quadrupole ion trap collisional activation conditions. In particular, fragmentation that requires two cleavages (i.e., cleavage of a disulfide linkage and a peptide linkage) is strongly suppressed. Reduction of the disulfide linkage(s) by use of dithiothreitol yields parent ions upon electrospray without this complication. Far richer structural information is revealed by ion trap collisional activation of the disulfide-reduced species than from the native species. These observations are illustrated with doubly protonated native and reduced somatosin, the [M + 5H](5+) ion of native bovine insulin and the [M + 4H](4+) and [M + 3H](3+) ions of the B-chain of bovine insulin produced by reduction of the disulfide linkages in insulin, and the [M + 11H](11+) ion of native chicken lysozyme and the [M + 11H](11+) and [M + 14H](14+) ions of reduced lysozyme. In each case, the product ions produced by ion trap collisional activation were subjected to ion/ion proton transfer reactions to facilitate interpretation of the product ion spectra. These studies clearly suggest that the identification of polypeptides with one or more disulfide linkages via application of ion trap collisional activation to the multiply charged parent ions formed directly by electrospray could be problematic. Means for cleaving the disulfide linkage, such as reduction by dithiothreitol prior to electrospray, are therefore desirable in these cases.

Amino Acid Sequence↗

Venomous adversaries: a reference to snake identification, field safety, and bite-victim first aid for disaster-response personnel deploying into the hurricane-prone regions of North America.

Each hurricane season, emergency-preparedness deployment teams including but not limited to the Office of Force Readiness and Deployment of the US Public Health Service, Federal Emergency Management Agency, Deployment Medical Assistance Teams, Veterinary Medical Assistance Teams, and the US Army and Air Force National Guard are at risk for deploying into hurricane-stricken areas that harbor indigenous hazards, including those posed by venomous snakes. North America is home to 2 distinct families of venomous snakes: 1) Viperidae, which includes the rattlesnakes, copperheads, and cottonmouths; and 2) Elapidae, in which the only native species are the coral snakes. Although some of these snakes are easily identified, some are not, and many rank among the most feared and misunderstood animals. This article specifically addresses all the native species of venomous snakes that inhabit the hurricane-prone regions of North America and is intended to serve as a reference to snake identification, basic field safety procedures, and the currently recommended first-aid measures for snakebite casualties.

Animals↗

Initial effects of light armored vehicle use on grassland vegetation at Fort Lewis, Washington.

Sustainable use of military training lands requires understanding and prediction of the effects of military vehicles on vegetation. We examined the initial impacts of an 8-wheeled, light armored vehicle (LAV) on grassland vegetation at Fort Lewis, Washington. The LAV drove replicate spiral paths at two starting velocities, 10.3 and 5.1 m s(-1). The disturbed width (width of ground impacted by the tires) increased as turning radius decreased, but was unaffected by vehicle velocity. An inverse-exponential model predicted disturbed width (r(2)=0.68) at all turning radii for both velocities combined. In low-velocity spirals, and for straight tracking (turning radius>40 m) and moderate turns (radius 20-40 m) in high-velocity spirals, all vegetation damage was imprint (plants flattened by wheels). During sharp (radius <20 m), high-velocity turns, most or all of the disturbed width was scraped free of surface vegetation and soil, which was piled to the outside of each tire track. Total plant cover (all species) was not affected by track curvature in low-velocity spirals, but decreased in the order straight tracking>moderate turns>sharp turns in high-velocity spirals. In low-velocity spirals, post-tracking cover of several plant growth forms (non-native species, perennial species, sod-forming grasses) was similar to pre-tracking cover, but in high-velocity spirals, post-tracking cover of these growth forms decreased in the order straight > or =moderate=sharp. Cover of native species and forbs decreased more in high- than in low-velocity spirals, but was unaffected by curvature. Pre- and post-tracking cover of annual species, bunchgrasses, and shrubs was < or =3%. The most severe vegetation damage caused by operation of wheeled LAVs on grasslands is associated with sharp, high-velocity turns.

Analysis of Variance↗

Biomarkers of a native fish species (Cnesterodon decemmaculatus) application to the water toxicity assessment of a peri-urban polluted river of Argentina.

Environmental monitoring by means of biomarker parameters assessed in different species is a useful tool. It has the advantage of providing a quantitative response as well as valuable information of ecological relevance on the chronic adverse effects caused by water pollution. The aim of this study was to assess the response of biochemical and physiological parameters of Cnesterodon decemmaculatus, a native teleost, simultaneously caught in two sites of Reconquista river, a highly polluted peri-urban river. This study compared the measured parameters with that of specimens of the same species captured in an unpolluted body water, and correlated the detected changes with the physicochemical profile of the water at each site. A comparison was made of selected parameters of gill, brain and liver and of somatic indices of fish collected from polluted and reference sites. The main parameters whose changes allowed to discriminate between sampling sites were gill (Na(+),K(+))-ATPase, brain acetylcholinesterase (AChE) and liver aminotransaminases activities; tissues' protein content and liver somatic index (LSI) were also sensitive biomarkers in brain and liver, respectively. The results showed that the response of the measured biomarkers allowed for the differentiation of sampling sites according to their water quality and confirmed that Cnesterodon decemmaculatus may be a useful test organism for the biomonitoring of freshwater environments. In addition, the simultaneous measurement of the physicochemical parameters of the water samples showed a good correspondence between the biomarkers responses and the environmental chemical stress conditions.

Acetylcholinesterase↗

Reactions to less common species of fire ants.

BACKGROUND: There are four species of fire ants found in the United States in addition to the most common, Solenopsis invicta. Reactions have been reported from stings of each of these species, but large numbers of insects and adequate amounts of venom for study are very difficult to obtain. METHODS: Venom was obtained, the purified allergens were isolated, and the complete amino acid sequences were determined for two of the three allergens from S. richteri. Skin testing and RAST studies were performed on patients with reactions to native fire ant stings and analyzed in comparison with clinical history. RESULTS: The structures of S. richteri allergens have a high degree of similarity to S. invicta allergens. The Sol 2 allergens are less related to each other than either the Sol 1 (phospholipase AB) or Sol 3 (antigen 5) allergens. Patients sensitized to native species of fire ants react primarily to the Sol 1 and Sol 3 allergens, whereas those originally sensitized to S. invicta also react significantly to the Sol 2 and Sol 4 allergens. Some patients are initially sensitized to S. invicta and have life-threatening reactions to stings of native species. The tropical fire ant, S. geminata, has become a serious problem in some areas of the Pacific and South Asia, especially Okinawa and Guam. CONCLUSIONS: The venoms from all of the species of fire ants examined appear to be highly cross-reactive. S. invicta imported fire ant venom extracts are probably sufficient for diagnosis and may warrant a clinical trial for immunotherapy of allergic reactions to venoms of any of the other four species.

Adult↗

Gelatin-sealed polyester resists Staphylococcus epidermidis biofilm infection.

BACKGROUND: The purpose of this study was to show that gelatin-impregnated polyester grafts inhibit Staphylococcus epidermidis biofilm infection in a canine model of aortic graft interposition. A clinically native species and two engineered strains, which differed in slime and adhesin antigen components, were compared to determine differential gelatin and slime interactions. METHODS: In vitro bacterial graft colonization was validated by immersion of graft segments in inoculating solutions (10(6) colony forming units/ml) of a clinically native species RP62A and two genetically engineered S. epidermidis species, M187sn3 (SN3: slime and adhesin negative) or M187sp11 (SP11: slime and adhesin positive), for 18 h at 23 degrees C. The grafts were washed, sonicated, and cultured to assess in vitro bacterial graft adherence. Grafts similarly inoculated were placed as aortic interposition grafts in dogs. Three sterile grafts were implanted as controls. Grafts were excised after 6 weeks and cultured for bacterial growth as in the in vitro study. Infection was defined by a positive culture in the excised grafts. Data were analyzed with nonparametric statistical methods. RESULTS: In vitro bacterial graft adherence in colony forming units per milliliter was similar at 18 h postsonication for RP62A (8 x 10(4) +/- 1 x 10(4)), SN3 (7 x 10(4) +/- 2 x 10(4)), and SP11 (6 x 10(4) +/- 2 x 10(4)) (P = NS). Only one of five grafts inoculated with RP62A was culture positive after 6 weeks. No grafts inoculated with the engineered strains SN3 or SP11 were culture positive after explanation. CONCLUSION: In vitro bacterial inoculation of gelatin-impregnated polyester was similar among the species and not dependent upon the presence of slime and adhesin components. Gelatin-impregnated polyester grafts demonstrated in vivo resistance to coagulase-negative staphylococcal biofilm infection.

Animals↗

Structure of the zona pellucida and cumulus oophorus in three species of native Australian rodents.

The sperm head of many Australian hydromyine rodents has three curved hooks projecting from its anterior margin; the structure of the hooks has been characterized, but their function is unknown. In this study, we have investigated whether the hooks might have evolved to assist sperm penetration through more formidable egg vestments, particularly the zona pellucida. Cumulus-oocyte complexes were obtained from two species that possess a three-hooked sperm head (Pseudomys australis and P. nanus) and one species that does not (Notomys alexis) and examined by light and electron microscopy. After fixation in the presence of ruthenium red, the zona pellucida was found to consist of a fibrillar meshwork, but there were no interspecific structural differences. A corona radiata was absent, and the cumulus extracellular matrix was composed of filaments and electron-dense granules in each species. Measurements of the zona thickness in freshly ovulated, unfixed oocytes revealed that it was thinnest (7.8 microns) in P. australis, which has a three-hooked sperm head, and thickest (11.4 microns) in N. alexis, the species in which the ventral hooks are absent. Hence, no correlation was found between the thickness of the zona pellucida or the structure of the cumulus-oocyte complex, and the presence of three hooks on the sperm head. We conclude, therefore, that it is unlikely that the evolution of the three-hooked sperm head is an adaptation for penetration of increased barriers around the oocyte.

Animals↗

Toxoplasmosis in three species of native and introduced Hawaiian birds.

Toxoplasma gondii was found in endemic Hawaiian birds, including 2 nene geese (Nesochen sandvicensis), 1 red-footed booby (Sula sula), and an introduced bird, the Erckels francolin (Francolinus erckelii). All 4 birds died of disseminated toxoplasmosis; the parasite was found in sections of many organs, and the diagnosis was confirmed by immunohistochemical staining with anti-T. gondii-specific polyclonal antibodies. This is the first report of toxoplasmosis in these species of birds.

Animals↗

Invasion of an asexual American water flea clone throughout Africa and rapid displacement of a native sibling species.

The huge ecological and economic impact of biological invasions creates an urgent need for knowledge of traits that make invading species successful and factors helping indigenous populations to resist displacement by invading species or genotypes. High genetic diversity is generally considered to be advantageous in both processes. Combined with sex, it allows rapid evolution and adaptation to changing environments. We combined paleogenetic analysis with continent-wide survey of genetic diversity at nuclear and mitochondrial loci to reconstruct the invasion history of a single asexual American water flea clone (hybrid Daphnia pulexxDaphnia pulicaria) in Africa. Within 60 years of the original introduction of this invader, it displaced the genetically diverse, sexual population of native D. pulex in Lake Naivasha (Kenya), despite a formidable numerical advantage of the local population and continuous replenishment from a large dormant egg bank. Currently, the invading clone has spread throughout the range of native African D. pulex, where it appears to be the only occurring genotype. The absence of genetic variation did not hamper either the continent-wide establishment of this exotic lineage or the effective displacement of an indigenous and genetically diverse sibling species.

Africa↗

[Chemical characteristics of the pulp and oil of the annona tree (Annona coriaceae)].

Belonging to the Annonaceae family, marolo (Annona coriaceae) is a native species of the Brazilian "cerrado" región (Minas Gerais, Goiás and Distrito Federal) and can be found in South American tropical zones. Its fruits are highly consumed by local people and commercialized in markets or street stalls. There is, however, a tendency for the extinction of marolo due to deforestation and the large scale plantation of monocultures instead of native plants. The literature still offers no data on the chemical composition of the proximate composition and vitamin C, A and tannin contents were carried out on the yellow marolo pulp as well as the determination of the physico-chemical characteristics of the seed oil. Five batches of fruit from the Alfenas region--south of Minas Gerais State--were analysed in this work and their average composition were: humidity 77%, total sugar 15%, reducing sugar 11%, crude protein 1%, lipids 3%, fiber 5% and fixed mineral residue 1%. The contents of vitamin C and A were 8.2 mg/100g and 117.5 RE/100g, respectively, and the tannin content was 245 mg/100g. The results showed high fiber and lipid contents of marolo pulp in comparison with many other tropical fruit pulps. The vitamin C contents were equivalent to those found in avocado, pineapple and watermelon, while the vitamin A contents were equivalent to papaya, peach, guava and several other tropical fruits. Marolo seed contains 45% of oil on a dry basis. Its composition and physico-chemical characteristics showed the possibility of producing a good quality oil, with great potential for the fine oil market. However the presence of alkaloids in the oil needs to be further studied. Their elimination could be done by refining or extraction in a continuous press. The results exalt the high quality of marolo pulp, showing that the preservation of native species should be stimulated.

Alkaloids↗

Modelling the impacts of two exotic invasive species on a native butterfly: top-down vs. bottom-up effects.

1. Exotic invasive species can influence population dynamics of native species through top-down or bottom-up forces. The present study examined separate and interactive effects of multiple exotic species invasions on the native mustard white butterfly, Pieris napi oleracea Harris (Lepidoptera: Pieridae), using a stochastic simulation model. 2. P. n. oleracea populations in North America have decreased regionally since the 1860s. Competition with an exotic congener (P. rapae L.), loss of native host plants and parasitism by the introduced broconid wasp (Cotesia glomerata L.), have been suggested to be independently responsible for its decline. The present study examined these hypotheses, as well as an alternative, invasion by an exotic crucifer, garlic mustard (Alliaria petiolata[Bieb.] Cavara & Grande). 3. A stochastic simulation model of P. n. oleracea population dynamics revealed that decreasing the number of host plants available for oviposition and larval development (i.e. habitat loss), sharply reduced the probability of populations persistence and decreased population size for those that persisted. 4. Simulated invasion by garlic mustard also substantially decreased both probability of persistence (= 0 at approximately 50% cover) and mean population size. Persistence probability never reached zero under any C. glomerata scenarios, even when larval mortality in the second generation due to parasitism was 100%. The impact of garlic mustard was intensified by the addition of C. glomerata parasitism. 5. Results suggest that bottom-up forces, loss of host plants through forest understorey loss and/or garlic mustard invasion are the most important forces driving P. n. oleracea population decline. Parasitism by C. glomerata may interact to reduce P. n. oleracea populations more rapidly, but appears insufficient alone to cause local extinction.

Animals↗