The Cloacal Microbiome Changes with the Breeding Season in a Wild Bird.

The symbiotic microbial communities, or "microbiomes," that reside on animals are dynamic, and can be affected by the behavior and physiology of the host. These communities provide many critical beneficial functions for their hosts, but they can also include potential pathogens. In birds, bacteria residing in the cloaca form a complex community, including both gut and sexually-transmitted bacteria. Transmission of cloacal bacteria among individuals is likely during the breeding season, when there is direct cloacal contact between individuals. In addition, the major energetic investment in reproduction can draw resources away from immune responses that might otherwise prevent the successful establishment of microbes. We assessed dynamic variation in the cloacal microbiome of free-living rufous-collared sparrows (Zonotrichia capensis) through sequential breeding and non-breeding seasons. We found that the cloacal bacterial communities differed between the sexes when they were in breeding condition. Further, in males, but not in females, the bacterial community became more diverse with the onset of reproduction, and then decreased in diversity as males transitioned to non-breeding condition. Individuals sampled across sequential breeding seasons did not accumulate more bacterial taxa over seasons, but bacterial community composition did change. Our results suggest that the cloacal microbiome in birds is dynamic and, especially in males, responsive to breeding condition.

Larval wood frog (Rana [=Lithobates] sylvatica) development and physiology following infection with the trematode parasite, Echinostoma trivolvis.

Parasites can potentially affect host energetics through a variety of mechanisms including diverting energy from host functions or eliciting energetically costly responses. In many systems energetic costs of parasite infection remain poorly defined. The widespread trematode Echinostoma trivolvis can cause mortality of and pathology in larval amphibians. However, physiological impacts of E. trivolvis infection have received limited attention. To evaluate the effects of E. trivolvis on larval amphibian survival, growth and development, we studied a wide range of infection intensity in wood frog, Rana (=Lithobates) sylvatica, tadpoles in laboratory experiments and outdoor mesocosms. To assess potential underlying physiological costs of infection, we measured tadpole energetics and phenotypic plasticity of the intestines as a compensatory mechanism to offset increased energy costs. Survival was high in all tadpoles, but the highest infections decreased the growth and slowed the development of tadpoles raised in mesocosms and the laboratory. However, infections failed to elicit detectable energetic costs or phenotypic changes in intestinal size. The lack of energetic costs observed in our study emphasizes the complex and often context-dependent nature of energetic costs of parasitism and suggests that other mechanisms, such as changes in host behavior, may contribute to sub-lethal effects on growth and development.

Glucocorticosteroid hormone treatment of larval treefrogs increases infection by Alaria sp. trematode cercariae.

In many amphibian species, an apparent increase has occurred in the prevalence of limb deformities caused by parasitic trematodes. We are interested in the role of environmental stressors in increasing these infections in amphibians. One mechanism by which environmental stressors could act to increase disease prevalence is to increase circulating levels of glucocorticosteroid hormones, which are released in response to stressors and can be immunosuppressive. In the present study, we treated gray treefroZg tadpoles (Hyla versicolor) with exogenous corticosterone, which is the main glucocorticosteroid "stress" hormone in amphibians. We then exposed treated tadpoles to Alaria sp. cercariae and scored the number of mesocercariae that successfully infected the tadpoles. In addition, we assayed one function of the immune response by counting the number of circulating eosinophilic granulocytes, which are thought to be important in immune responses to macroparasites. Tadpoles treated with exogenous corticosterone developed higher parasite loads than control tadpoles did, and they had lower numbers of circulating eosinophilic granulocytes. These results provide evidence of glucocorticosteroid-mediated immunosuppression in tadpoles that may help to explain apparent increases in the numbers of trematode-induced deformities in amphibian populations during recent decades.

Sodium reduction for hypertension prevention in overweight adults: further results from the Trials of Hypertension Prevention Phase II.

Sodium reduction is efficacious for primary prevention of hypertension, but the feasibility of achieving this effect is unclear. The objective of the paper is detailed analyses of adherence to and effects of the sodium reduction intervention among overweight adults in the Trials of Hypertension Prevention, Phase II. Sodium reduction (comprehensive education and counselling about how to reduce sodium intake) was tested vs no dietary intervention (usual care) for 36-48 months. A total of 956 white and 203 black adults, ages 30-54 years, with diastolic blood pressure 83-89 mmHg, systolic blood pressure (SBP) <140 mmHg, and body weight 110-165% of gender-specific standard weight were included in the study. At 36 months, urinary sodium excretion was 40.4 mmol/24 h (24.4%) lower in sodium reduction compared to usual care participants (P<0.0001), but only 21% of sodium reduction participants achieved the targeted level of sodium excretion below 80 mmol/24 h. Adherence was positively related to attendance at face-to-face contacts. Net decreases in SBP at 6, 18, and 36 months of 2.9 (P<0.001), 2.0 (P<0.001), and 1.3 (P=0.02) mmHg in sodium reduction vs usual care were associated with an overall 18% lower incidence of hypertension (P=0.048); were relatively unchanged by adjustment for ethnicity, gender, age, and baseline blood pressure, BMI, and sodium excretion; and were observed in both black and white men and women. From these beneficial but modest results with highly motivated and extensively counselled individuals, sodium reduction sufficient to favourably influence the population blood pressure distribution will be difficult to achieve without food supply changes.

Juvenile amphibians do not avoid potentially lethal levels of urea on soil substrate.

We examined the effects of a forest fertilizer (urea) on newly metamorphosed terrestrial amphibians (Western toads, Bufo boreas; Cascades frogs, Rana cascadae; long-toed salamanders, Ambystoma macrodactylum; and roughskin newts, Taricha granulosa). We examined avoidance behavior of Western toads and Cascades frogs on both paper towel and soil substrates dosed with urea (control and 100 kg N/ha and an additional treatment of 50 kg N/ha for Western toads on soil substrate) and avoidance behavior of long-toed salamanders on soil substrate dosed with urea. We further examined the survival and feeding behavior of all four species exposed to urea on soil substrate (100 kg N/ha) for 5 d. Juvenile Western toads and Cascades frogs avoided paper towels dosed with urea but did not avoid urea-dosed soil substrate. However, Western toads and Cascades frogs both suffered significant mortality when exposed to urea on a soil substrate for 5 d. Furthermore, after adjusting for weight, we found that urea-exposed juvenile Western toads and Cascades frogs consumed significantly fewer prey items (crickets) compared with nonexposed control animals. Long-toed salamanders did not discriminate against soil substrate dosed with urea, and neither long-toed salamanders nor roughskin newts died or reduced prey consumption as a result of urea exposure. Juvenile amphibians may not be able to detect and avoid harmful levels of urea fertilizer on a natural substrate. Furthermore, anthropogenic stressors such as urea fertilizer can significantly reduce the survival and prey consumption of juvenile amphibians. These effects are important to consider in light of possible threats to the conservation status of many amphibian species.

Sensitivity to urea fertilization in three amphibian species.

Forest fertilization with granular urea is a well-established management practice in many forested regions of the world. We hypothesize that chemical forest fertilizers may be affecting forest-dwelling wildlife. In the laboratory, we studied the effects of fertilization doses of granular urea on three species of forest-dwelling amphibians (Plethodon vehiculum, Rhyacotriton variegatus, and Taricha granulosa). In avoidance experiments, the three species avoided a substrate treated with a dose of 225 kg N/ha urea. In toxicity experiments, we exposed amphibians to urea at doses of 225 kg N/ha and 450 kg N/ha for 4 days. The observed effects increased with time and dose, and there were significant differences in sensitivity among the species. Both treatment levels had an acute effect on survival of P. vehiculum and R. variegatus. At 24 h, mortality at the highest dose was 67% for P. vehiculum, and 47% for R. variegatus. In contrast, there was no mortality for T. granulosa at these concentrations. We suggest that environmental levels of urea could be affecting behavior and survival of some amphibians species in fertilized forests.

Complex causes of amphibian population declines.

Amphibian populations have suffered widespread declines and extinctions in recent decades. Although climatic changes, increased exposure to ultraviolet-B (UV-B) radiation and increased prevalence of disease have all been implicated at particular localities, the importance of global environmental change remains unclear. Here we report that pathogen outbreaks in amphibian populations in the western USA are linked to climate-induced changes in UV-B exposure. Using long-term observational data and a field experiment, we examine patterns among interannual variability in precipitation, UV-B exposure and infection by a pathogenic oomycete, Saprolegnia ferax. Our findings indicate that climate-induced reductions in water depth at oviposition sites have caused high mortality of embryos by increasing their exposure to UV-B radiation and, consequently, their vulnerability to infection. Precipitation, and thus water depth/UV-B exposure, is strongly linked to El Niño/Southern Oscillation cycles, underscoring the role of large-scale climatic patterns involving the tropical Pacific. Elevated sea-surface temperatures in this region since the mid-1970s, which have affected the climate over much of the world, could be the precursor for pathogen-mediated amphibian declines in many regions.

Long-term weight loss and changes in blood pressure: results of the Trials of Hypertension Prevention, phase II.

BACKGROUND: Weight loss appears to be an effective method for primary prevention of hypertension. However, the long-term effects of weight loss on blood pressure have not been extensively studied. OBJECTIVE: To present detailed results from the weight loss arm of Trials of Hypertension Prevention (TOHP) II. DESIGN: Multicenter, randomized dinical trial testing the efficacy of lifestyle interventions for reducing blood pressure over 3 to 4 years. Participants in TOHP II were randomly assigned to one of four groups. This report focuses only on participants assigned to the weight loss (n = 595) and usual care control (n = 596) groups. PATIENTS: Men and women 30 to 54 years of age who had nonmedicated diastolic blood pressure of 83 to 89 mm Hg and systolic blood pressure less than 140 mm Hg and were 110% to 165% of their ideal body weight at baseline. INTERVENTION: The weight loss intervention included a 3-year program of group meetings and individual counseling focused on dietary change, physical activity, and social support MEASUREMENTS: Weight and blood pressure data were collected every 6 months by staff who were blinded to treatment assignment RESULTS: Mean weight change from baseline in the intervention group was -4.4 kg at 6 months, -2.0 kg at 18 months, and -0.2 kg at 36 months. Mean weight change in the control group at the same time points was 0.1, 0.7, and 1.8 kg. Blood pressure was significantly lower in the intervention group than in the control group at 6, 18, and 36 months. The risk ratio for hypertension in the intervention group was 0.58 (95% CI, 0.36 to 0.94) at 6 months, 0.78 (CI, 0.62 to 1.00) at 18 months, and 0.81 (CI, 0.70 to 0.95) at 36 months. In subgroup analyses, intervention participants who lost at least 4.5 kg at 6 months and maintained this weight reduction for the next 30 months had the greatest reduction in blood pressure and a relative risk for hypertension of 0.35 (CI, 0.20 to 0.59). CONCLUSIONS: Clinically significant long-term reductions in blood pressure and reduced risk for hypertension can be achieved with even modest weight loss.

Trials of Hypertension Prevention, phase II. Structure and content of the weight loss and dietary sodium reduction interventions. Trials of Hypertension Prevention (TOHP) Collaborative Research Group.

Identifying effective, nonpharmacologic means of preventing or significantly delaying the onset of hypertension would be a major advance in the primary prevention of cardiovascular disease. In the first phase of the Trials of Hypertension Prevention (TOHP I), adults with high-normal diastolic blood pressure were randomly assigned to one of seven nonpharmacologic interventions. Only weight loss and reduction of dietary sodium proved to be effective strategies for reducing blood pressure. The second phase of TOHP (TOHP II) will test the effectiveness of weight loss, reduction of dietary sodium, and their combination of lowering blood pressure and preventing the onset of hypertension over a 3- to 4-year follow-up period. This article describes the three interventions used in TOHP II, methods used to maintain continued participation in this long-term trial, and protocol enhancements designed to maximize intervention effectiveness.