Thermal biology, activity, and population parameters of Cnemidophorus vacariensis (Squamata, Teiidae), a lizard endemic to southern Brazil

We investigated the following aspects of the biology of a population of Cnemidophorus vacariensis Feltrim & Lema, 2000 during the four seasons: thermal biology, relationship with the thermal environment, daily and seasonal activity, population structure and growth rate. Cnemidophorus vacariensis is restricted to rocky outcrops of the "campos de cima da serra" grasslands on the Araucaria Plateau, southern Brazil, and is currently listed as regionally and nationally threatened with extinction. Data were collected from October 2004 through September 2007 in the state of Rio Grande do Sul. Sampling was conducted randomly from 08:00 a.m. to 6:00 p.m. The capture-mark-recapture method was employed. The lizards were captured by hand, and their cloacal temperature, sex, snout-ventral length (SVL), mass, and the temperature of their microhabitat (substrate temperature and air temperature) were recorded. Individuals were then marked by toe-clipping and released at the site of capture. Body temperatures were obtained for 175 individuals, activity data for 96 individuals, and data on population structure and growth for 59 individuals. All data were obtained monthly, at different times of the day. Cnemidophorus vacariensis average body temperature was 23.84ºC, ranging between 9.6 and 38.2ºC. Temperatures ranged between 21 and 29ºC. The correlation between external heat sources, substrate and air were positive and significant and there was a greater correlation between lizard's temperature and the temperature of the substrate (tigmothermic species). The relatively low body temperatures of individuals are associated with the climate of their environment (altitude up to 1,400 m), with large variations in temperature throughout the day and the year, and low temperatures in winter. The average body temperature observed for C. vacariensis was low when compared with that of phylogenetically related species, suggesting that the thermal biology of this species reflects adaptations to the temperate region where it lives. The monthly rates of activity of lizards were related to monthly variations in the ambient temperatures. Our data suggest that the daily and seasonal activity of C. vacariensis result from the interaction between two factors: changes in the environment temperature and the relationship between individuals and their thermal environment. The population structure of C. vacariensis varied throughout the study period, with maximum biomass in January and maximum density in February (recruitment period). The sex ratio diverged from the expected 1:1. The growth analysis showed a negative relationship between the growth rate of individuals and the SVL, revealing that young individuals grow faster than adults, a typical pattern for short-lived species. The population studied showed a seasonal and cyclical variation associated with the reproductive cycle. The life strategy of C. vacariensis seems to include adaptations to the seasonal variations in temperature, typical of its environment.

Synchaeta jollyae (Shiel & Koste) (Rotifera, Synchaetidae): an overlooked species or a newcomer in Neotropical freshwaters?

Populations of Synchaeta jollyae (Shiel & Koste, 1993) (Rotifera), a species recently recorded for the first time in Brazil and South America, were analyzed in reservoirs in Southeast Brazil. Sampling was carried out monthly from August 2006 to July 2007 at Furnas Reservoir in the Rio Grande basin, state of Minas Gerais, and in four cascade reservoirs in the Tietê River basin (Barra Bonita, Bariri, Ibitinga and Nova Avanhandava) state of São Paulo, in June and September 2008 and in January and May 2009. Synchaeta jollyae occurred in most samples and periods. From the results obtained it is evident that S. jollyae occurs in water bodies of varied trophic status but reaches larger populations in eutrophic water bodies and during lower temperature periods. The greatest densities of S. jollyae were found in the eutrophic Bariri Reservoir, on the Tietê River, during the winter. Mann-Whitney test confirmed the significant difference between the population densities in periods of high and low temperatures, with populations reaching higher densities at lower temperatures. It is not yet possible to tell whether S. jollyae is a widely distributed species that has been overlooked in previous plankton studies in South America. Wherever these populations of S. jollyae might have originated, it appears to be a species well established and adapted to a wide range of conditions in the Neotropics.

Properties of ß-lactamase from Neisseria gonorrhoeae

ß-lactamase activity was studied in Neisseria gonorrhoeae strains. Optimum temperature was found to be 37°C. The enzyme was inactivated at temperatures higher than 60°C, but remained active during storage at low temperatures (4°C, -30°C and -70°C) for two months. Enzyme activity was observed within a pH range of 5.8-8.0, while the optimum pH was 7.0-7.2. Addition of Ni2+, Fe2+, Fe3+, Mn2+ and p-chloromercurybenzoate to the reaction buffer exerted a negative effect upon the activity, whereas Hg2+ and ethylene diamine tetra-acetic acid produced complete inhibition. These results would indicate the presence of -SH groups at the catalytic site of the enzyme.

Effect of Sequential Cold Shocks on Survival and Molting Rate in Triatoma infestans Klug

The survival and molting incidence in Triatoma infestans, a vector of Chagas disease, were investigated following sequential shocks at 0ºC in fifth instar nymphs under moderate fasting and full nutritional conditions. The shocks were separated by intervals of 8 h and 24 h at 30ºC. The results indicated that in terms of insect survival, T. infestans is tolerant to a single cold shock at 0ºC even for 12 h, or to sequential cold shocks, regardless of the nutritional state of the specimens. In terms of molting rate, fasting enhanced the tolerance to sequential cold shocks, but did not exceed the tolerance acquired by fully-nourished specimens, except when cold shocks were separated by an 8 h interval at 30ºC. The protective action elicited by fasting was assumed to be additive to that induced by a single mild cold shock or sequential cold shocks. The cold-tolerance response of T. infestans may have favoured its survival in areas of South America with low temperatures, even considering that this species is predominantly associated with human habitats.

Exogenous selection shapes germination behaviour and seedling traits of populations at different altitudes in a Senecio hybrid zone.

BACKGROUND AND AIMS: The Senecio hybrid zone on Mt Etna, Sicily, is characterized by steep altitudinal clines in quantitative traits and genetic variation. Such clines are thought to be maintained by a combination of 'endogenous' selection arising from genetic incompatibilities and environment-dependent 'exogenous' selection leading to local adaptation. Here, the hypothesis was tested that local adaptation to the altitudinal temperature gradient contributes to maintaining divergence between the parental species, S. chrysanthemifolius and S. aethnensis. METHODS: Intra- and inter-population crosses were performed between five populations from across the hybrid zone and the germination and early seedling growth of the progeny were assessed. KEY RESULTS: Seedlings from higher-altitude populations germinated better under low temperatures (9-13 °C) than those from lower altitude populations. Seedlings from higher-altitude populations had lower survival rates under warm conditions (25/15 °C) than those from lower altitude populations, but also attained greater biomass. There was no altitudinal variation in growth or survival under cold conditions (15/5 °C). Population-level plasticity increased with altitude. Germination, growth and survival of natural hybrids and experimentally generated F(1)s generally exceeded the worse-performing parent. CONCLUSIONS: Limited evidence was found for endogenous selection against hybrids but relatively clear evidence was found for divergence in seed and seedling traits, which is probably adaptive. The combination of low-temperature germination and faster growth in warm conditions might enable high-altitude S. aethnensis to maximize its growth during a shorter growing season, while the slower growth of S. chrysanthemifolius may be an adaptation to drought stress at low altitudes. This study indicates that temperature gradients are likely to be an important environmental factor generating and maintaining adaptive divergence across the Senecio hybrid zone on Mt Etna.

Sensitivity and specificity of polymerase chain reaction in Giemsa-stained slides for diagnosis of visceral leishmaniasis in children

The aim of this study was to evaluate the sensitivity and specificity of polymerase chain reaction (PCR) in the detection of Leishmania DNA in archived Giemsa-stained bone marrow slides for diagnosis of visceral leishmaniasis (VL), and to compare PCR with conventional diagnostic techniques, like direct microscopy and parasite culture. Specimens of archived Giemsa-stained bone marrow slides from 91 patients with VL and from 79 controls with other diseases or conditions were studied. PCR showed the highest sensitivity (92.3%) and had good specificity (97.5%). Direct examination detected 79.1% and culture 59% of positive samples. In addition, PCR was able to detect VL in 16 of 19 patients (84.2%) with negative microscopy. PCR in Giemsa-stained bone marrow slides is a suitable tool for confirming diagnosis in patients with VL and may be useful in the diagnosis of difficult cases. Slide smears are easily stored, do not require special storage conditions such as low temperatures, and can be easily mailed to centers where PCR is available, making it an excellent option for diagnosis in the field.

Long-term storage and viability of Ochlerotatus albifasciatus (Diptera: Culicidae)

The viability of Ochlerotatus albifasciatus (Macquart) eggs stored at room temperature and at 5ºC was studied over 31 months. After 12, 18 and 31 months of storage, eggs were acclimatized at 22ºC for ten days, and then inundated twice every seven days. The effect of the storage period on the percentage of hatching was analyzed by one way ANOVA. Differences on the hatching response between the first and second flooding were analyzed by paired t-test. Differences on the hatching response between the two storage conditions were analyzed by Mann-Whitney rank test. Results showed that (1) Oc. albifasciatus eggs were able to survive and hatch over 31 months; (2) the percent hatching of eggs stored at 5ºC was higher than that of eggs stored at room temperature; and (3) low temperatures and long periods without water favor installment hatching.

Diversity of Chromobacterium violaceum isolates from aquatic environments of state of Pará, Brazilian Amazon

The present study intended to characterize the phenotypic and genetic diversity of Brazilian isolates of Chromobacterium violaceum from aquatic environments within the Amazon region. Nineteen isolates showed morphological properties of C. violaceum and the majority grew at 44°C. Low temperatures, in contrast, showed to be inhibitory to their growth, as eleven isolates did not grow at 10ºC and nine did not produce pigmentation, clearly indicating an inhibition of their metabolism. The largest variation among isolates was observed in the citrate test (Simmons), in which 12 isolates were positive, and in the oxidation/fermentation of sucrose, with six positives isolates. Chloramphenicol, gentamicin and sulfonamides efficiently inhibited bacterial growth. Amplified products of the recA gene were digested with HindII or PstI, which produced three or four restriction fragments patterns, respectively. The combined analysis arranged the isolates into six genospecies. The higher diversity observed in Belém (genotypes C, D, E and F) may be a consequence of intense human occupation, pollution of the aquatic environment or due to the higher diversity of the environments sampled in that region. In conclusion, a high level of genetic and phenotypic diversity was observed, and four new genospecies were described.

Effect of Temperature on the Development and Survival of the Argentine Ant, Linepithema humile

The influence of temperature on the developmental times and survival of insects can largely determine their distribution. For invasive species, like the Argentine ant, Linepithema humile Mayr (Hymenoptera: Formicidae), these data are essential for predicting their potential range based on mechanistic models. In the case of this species, such data are too scarce and incomplete to make accurate predictions based on its physiological needs. This research provides comprehensive new data about brood survival and developmental times at a wide range of temperatures under laboratory conditions. Temperature affected both the complete brood development from egg to adult worker and each of the immature stages separately. The higher the temperature, the shorter the development times. Brood survival from egg to adult was low, with the maximum survival rate being only 16% at 26º C. Temperature also affected survival of each of the immature stages differently: eggs were negatively affected by high temperatures, while larvae were negatively affected by low temperatures, and the survival of pupae was apparentlyindependent of environmental temperature. At 32º C no eggs survived, while at 18º C less than 2% of the eggs hatched into larva. The data from the present study are essential for developing prediction models about the distribution range of this tramp species based on its physiological needs in relation to temperature

Tuberculosis in migrant populations. A systematic review of the qualitative literature.

BACKGROUND The re-emergence of tuberculosis (TB) in low-incidence countries and its disproportionate burden on immigrants is a public health concern posing specific social and ethical challenges. This review explores perceptions, knowledge, attitudes and treatment adherence behaviour relating to TB and their social implications as reported in the qualitative literature. METHODS Systematic review in four electronic databases. Findings from thirty selected studies extracted, tabulated, compared and synthesized. FINDINGS TB was attributed to many non-exclusive causes including air-born transmission of bacteria, genetics, malnutrition, excessive work, irresponsible lifestyles, casual contact with infected persons or objects; and exposure to low temperatures, dirt, stress and witchcraft. Perceived as curable but potentially lethal and highly contagious, there was confusion around a condition surrounded by fears. A range of economic, legislative, cultural, social and health system barriers could delay treatment seeking. Fears of deportation and having contacts traced could prevent individuals from seeking medical assistance. Once on treatment, family support and "the personal touch" of health providers emerged as key factors facilitating adherence. The concept of latent infection was difficult to comprehend and while TB screening was often seen as a socially responsible act, it could be perceived as discriminatory. Immigration and the infectiousness of TB mutually reinforced each another exacerbating stigma. This was further aggravated by indirect costs such as losing a job, being evicted by a landlord or not being able to attend school. CONCLUSIONS Understanding immigrants' views of TB and the obstacles that they face when accessing the health system and adhering to a treatment programme-taking into consideration their previous experiences at countries of origin as well as the social, economic and legislative context in which they live at host countries- has an important role and should be considered in the design, evaluation and adaptation of programmes.

Effect of Temperature on the Development and Survival of the Argentine Ant, Linepithema humile

The influence of temperature on the developmental times and survival of insects can largely determine their distribution. For invasive species, like the Argentine ant, Linepithema humile Mayr (Hymenoptera: Formicidae), these data are essential for predicting their potential range based on mechanistic models. In the case of this species, such data are too scarce and incomplete to make accurate predictions based on its physiological needs. This research provides comprehensive new data about brood survival and developmental times at a wide range of temperatures under laboratory conditions. Temperature affected both the complete brood development from egg to adult worker and each of the immature stages separately. The higher the temperature, the shorter the development times. Brood survival from egg to adult was low, with the maximum survival rate being only 16% at 26° C. Temperature also affected survival of each of the immature stages differently: eggs were negatively affected by high temperatures, while larvae were negatively affected by low temperatures, and the survival of pupae was apparently independent of environmental temperature. At 32° C no eggs survived, while at 18° C less than 2% of the eggs hatched into larva. The data from the present study are essential for developing prediction models about the distribution range of this tramp species based on its physiological needs in relation to temperature

New Radicals and new applications of Dynamic Nuclear Polarization of biological systems

Dynamic Nuclear Polarization (DNP) is an emerging technique that could revolutionize the NMR study of small molecules at very low concentrations by the increase in sensitivity that results from transfer of polarization between electronic and nuclear spins. Although the underlying physics has been known for a long time, in the last few years there has been a lot of excitement on the chemistry and biology NMR community caused by the demonstration that the highly polarized nuclei that are prepared in solid state at very low temperatures (1-2 K) could be rapidly transferred to liquid samples at room temperature and studied in solution by conventional NMR techniques. In favorable cases several order of magnitude increases in sensitivity have been achieved. The technique is now mature enough that a commercial instrument is available. The efficiency of DNP depends on two crucial aspects: i) the efficiency of the nuclear polarization process and ii) the efficiency of the transfer from the initial solid state to the fluid state in which NMR is measured. The preferred areas of application (iii) will be dictated by situations in which the low concentration of the sample or its intrinsic low receptivity are the limiting factors .

Phosphatidylethanolamine critically supports internalization of cell-penetrating protein C inhibitor.

Although their contribution remains unclear, lipids may facilitate noncanonical routes of protein internalization into cells such as those used by cell-penetrating proteins. We show that protein C inhibitor (PCI), a serine protease inhibitor (serpin), rapidly transverses the plasma membrane, which persists at low temperatures and enables its nuclear targeting in vitro and in vivo. Cell membrane translocation of PCI necessarily requires phosphatidylethanolamine (PE). In parallel, PCI acts as a lipid transferase for PE. The internalized serpin promotes phagocytosis of bacteria, thus suggesting a function in host defense. Membrane insertion of PCI depends on the conical shape of PE and is associated with the formation of restricted aqueous compartments within the membrane. Gain- and loss-of-function mutations indicate that the transmembrane passage of PCI requires a branched cavity between its helices H and D, which, according to docking studies, precisely accommodates PE. Our findings show that its specific shape enables cell surface PE to drive plasma membrane translocation of cell-penetrating PCI.

Proteomic analysis of peach fruit mesocarp softening and chilling injury using difference gel electrophoresis (DIGE)

Background: Peach fruit undergoes a rapid softening process that involves a number of metabolic changes. Storing fruit at low temperatures has been widely used to extend its postharvest life. However, this leads to undesired changes, such as mealiness and browning, which affect the quality of the fruit. In this study, a 2-D DIGE approach was designed to screen for differentially accumulated proteins in peach fruit during normal softening as well as under conditions that led to fruit chilling injury. Results:The analysis allowed us to identify 43 spots -representing about 18% of the total number analyzed- that show statistically significant changes. Thirty-nine of the proteins could be identified by mass spectrometry. Some of the proteins that changed during postharvest had been related to peach fruit ripening and cold stress in the past. However, we identified other proteins that had not been linked to these processes. A graphical display of the relationship between the differentially accumulated proteins was obtained using pairwise average-linkage cluster analysis and principal component analysis. Proteins such as endopolygalacturonase, catalase, NADP-dependent isocitrate dehydrogenase, pectin methylesterase and dehydrins were found to be very important for distinguishing between healthy and chill injured fruit. A categorization of the differentially accumulated proteins was performed using Gene Ontology annotation. The results showed that the 'response to stress', 'cellular homeostasis', 'metabolism of carbohydrates' and 'amino acid metabolism' biological processes were affected the most during the postharvest. Conclusions: Using a comparative proteomic approach with 2-D DIGE allowed us to identify proteins that showed stage-specific changes in their accumulation pattern. Several proteins that are related to response to stress, cellular homeostasis, cellular component organization and carbohydrate metabolism were detected as being differentially accumulated. Finally, a significant proportion of the proteins identified had not been associated with softening, cold storage or chilling injury-altered fruit before; thus, comparative proteomics has proven to be a valuable tool for understanding fruit softening and postharvest.

Loss of interactions with ants under cold climate in a regional myrmecophilous butterfly fauna

Aim Specialized mutualistic clades may revert and thus increase their autonomy and generalist characteristics. However, our understanding of the drivers that trigger reductions in mutualistic traits and of the consequences for the tolerance of these species to various environmental conditions remains limited. This study investigates the relationship between the environmental niche and the degree of myrmecophily (i.e. the ability to interact with ants) among members of the Lycaenidae. Location The western Swiss Alps. Methods We measured the tolerance of Lycaenidae species to low temperatures by comparing observations from a random stratified field sampling with climatic maps. We then compared the species-specific degree of myrmecophily with the species range limits at colder temperatures while controlling for phylogenetic dependence. We further evaluated whether the community-averaged degree of myrmecophily increases with temperature, as would be expected in the case of environmental filters acting on myrmecophilous species. Results Twenty-nine Lycaenidae species were found during sampling. Ancestral state reconstruction indicated that the 24 species of Polyommatinae displayed both strong myrmecophily and secondary loss of mutualism; these species were used in the subsequent statistical analyses. Species with a higher degree of ant interaction were, on average, more likely to inhabit warmer sites. Species inhabiting the coldest environments displayed little or no interaction with ants. Main conclusions Colder climates at high elevations filter out species with a high degree of myrmecophily and may have been the direct evolutionary force that promoted the loss of mutualism. A larger taxon sampling across the Holarctic may help to distinguish between the ecological and evolutionary effects of climate.