No evidence of metabolic costs following adaptive immune activation or reactivation in house sparrows.

The energy cost of adaptive immune activation in endotherms is typically quantified from changes in resting metabolic rate following exposure to a novel antigen. An implicit assumption of this technique is that all variation in energy costs following antigenic challenge is due solely to adaptive immunity, while ignoring potential changes in the energy demands of ongoing bodily functions. We critically assess this assumption by measuring both basal metabolic rate (BMR) and exercise-induced maximal metabolic rate (MMR) in house sparrows before and after the primary and two subsequent vaccinations with either saline (sham) or two novel antigens (keyhole limpet haemocyanin and sheep red blood cells; KLH and SRBC, respectively). We also examined the effect of inducing male breeding levels of testosterone (T) on immune responses and their metabolic costs in both males and females. Although there was a moderate decrease in KLH antibody formation in T-treated birds, there was no effect of T on BMR, MMR or immunity to SRBC. There was no effect of vaccination on BMR but, surprisingly, all vaccinated birds maintained MMR better than sham-treated birds as the experiment progressed. Our findings caution against emphasizing energy costs or nutrient diversion as being responsible for reported fitness reductions following activation of adaptive immunity.

Lack of seasonal and moult-related stress modulation in an opportunistically breeding bird: The white-plumed honeyeater (Lichenostomus penicillatus).

This article is part of a Special Issue "SBN 2014". In most vertebrate species, glucocorticoid levels and stress sensitivity vary in relation to season and life-history stage. In birds, baseline corticosterone (CORT) and stress sensitivity are typically highest while breeding and decrease substantially during moult. Because elevated CORT adversely affects protein synthesis, moult-related CORT suppression is thought to be necessary for forming high-quality feathers. Surprisingly, some passerine species lack moult-related CORT suppression, but these are distinguished by having slow rates of moult and being opportunistic breeders. We examined baseline and stress-induced CORT levels in an opportunistically breeding Australian passerine, the white-plumed honeyeater (Lichenostomus penicillatus). Although this species has a slower moult rate than high-latitude breeders, it differs little from north-temperate passerines. Neither baseline nor stress-induced CORT levels varied with season (winter, spring or summer), sex or moult status in adult birds. While breeding tended to be highest in early spring through late summer, laparotomies revealed only limited reduction in testicular size in males the year round. In all but one sampling period, at least some females displayed follicular hierarchy. Breeding usually coincides with outbreaks of phytophagous insects, which can happen at any time of the year. This results in moult/breeding overlap when infestations occur in late spring or summer. The ability of this species to moult and breed at the same time while having breeding-levels of CORT demonstrates that CORT suppression is not a prerequisite for synthesis of high-quality feathers. An experimental design incorporating moulting and non-moulting phenotypes is suggested to test the functional significance of CORT suppression in other species.

Covariation in life-history traits: differential effects of diet on condition, hormones, behavior, and reproduction in genetic finch morphs.

The relative contribution of genetic and environmental factors in determining variation in life-history traits is of central interest to evolutionary biologists, but the physiological mechanisms underlying these traits are still poorly understood. Here we experimentally demonstrate opposing effects of nutritional stress on immune function, endocrine physiology, parental care, and reproduction between red and black head-color morphs of the Gouldian finch (Erythrura gouldiae). Although the body condition of black morphs was largely unaffected by diet manipulation, red birds were highly sensitive to dietary changes, exhibiting considerable within-individual changes in condition and immune function. Consequently, nutritionally stressed red birds delayed breeding, produced smaller broods, and reared fewer and lower-quality foster offspring than black morphs. Differences in offspring quality were largely due to morph-specific differences in parental effort: red morphs reduced parental provisioning, whereas black morphs adaptively elevated their provisioning effort to meet the increased nutritional demands of their foster brood. Nutritionally stressed genetic morphs also exhibited divergent glucocorticoid responses. Black morphs showed reduced corticosterone-binding globulin (CBG) concentrations and increased levels of free corticosterone, whereas red morphs exhibited reduced free corticosterone levels and elevated CBG concentrations. These opposing glucocorticoid responses highlight intrinsic differences in endocrine sensitivities and plasticity between genetic morphs, which may underlie the morph-specific differences in condition, behavior, and reproduction and thus ultimately contribute to the evolution and maintenance of color polymorphism.

Adverse effects of fipronil on avian reproduction and development: maternal transfer of fipronil to eggs in zebra finch Taeniopygia guttata and in ovo exposure in chickens Gallus domesticus.

Two studies were carried out to examine the impact of maternal fipronil exposure on embryonic and offspring development. In the first study, breeding female zebra finches were orally dosed with single sublethal levels of fipronil (1, 5, and 10 mg/kg body weight) to determine behavioural and developmental consequences on chicks following maternal pesticide exposure. Significant levels of fipronil and fipronil-sulfone residues were detected in eggs laid by females in all dosed groups, however, these were undetectable in eggs laid 13 days after treatment. The level of sulfone detected in eggs was consistently higher than that of the parent fipronil compound. Of the seven eggs laid in the treatment groups, only one (14%) chick hatched and this was from the lowest dose group. This chick was severely underdeveloped at 10 days of age in comparison to control chicks and fiproles were detected in brain, liver, and adipose tissues collected following euthanasia of this individual. In contrast, there was 100% hatchability of control group eggs and all chicks fledged nests on schedule. In the second study, domestic chicken eggs were injected with 5.5, 17.5, and 37.5 mg/kg egg weight of fipronil directly into the yolk sac on day 12 of incubation. Treatment did not affect hatching success, however, behavioural and developmental abnormalities were observed in hatchlings from the highest dose group. These chicks also demonstrated reduced feeding rates, as indicated by reduced body mass at 48 h period post hatch. Both fipronil and fipronil-sulfone residues were detected in brain and liver tissue of hatchlings at all pesticide dose levels tested.

Predicting avian distributions to evaluate spatiotemporal overlap with locust control operations in eastern Australia.

Locusts and grasshoppers cause considerable economic damage to agriculture worldwide. The Australian Plague Locust Commission uses multiple pesticides to control locusts in eastern Australia. Avian exposure to agricultural pesticides is of conservation concern, especially in the case of rare and threatened species. The aim of this study was to evaluate the probability of pesticide exposure of native avian species during operational locust control based on knowledge of species occurrence in areas and times of application. Using presence-absence data provided by the Birds Australia Atlas for 1998 to 2002, we developed a series of generalized linear models to predict avian occurrences on a monthly basis in 0.5 degrees grid cells for 280 species over 2 million km2 in eastern Australia. We constructed species-specific models relating occupancy patterns to survey date and location, rainfall, and derived habitat preference. Model complexity depended on the number of observations available. Model output was the probability of occurrence for each species at times and locations of past locust control operations within the 5-year study period. Given the high spatiotemporal variability of locust control events, the variability in predicted bird species presence was high, with 108 of the total 280 species being included at least once in the top 20 predicted species for individual space-time events. The models were evaluated using field surveys collected between 2000 and 2005, at sites with and without locust outbreaks. Model strength varied among species. Some species were under- or over-predicted as times and locations of interest typically did not correspond to those in the prediction data set and certain species were likely attracted to locusts as a food source. Field surveys demonstrated the utility of the spatially explicit species lists derived from the models but also identified the presence of a number of previously unanticipated species. These results also emphasize the need for special consideration of rare and threatened species that are poorly predicted by presence-absence models. This modeling exercise was a useful a priori approach in species risk assessments to identify species present at times and locations of locust control applications, and to discover gaps in our knowledge and need for further focused data collection.

The effect of acute fenitrothion exposure on a variety of physiological indices, including avian aerobic metabolism during exercise and cold exposure.

The effect of fenitrothion exposure on birds was examined by measuring aerobic metabolism, blood hemoglobin content, plasma cholinesterases, and body weight for up to 21 d postdose. Peak metabolic rate was measured in a flight chamber in three-dose groups of house sparrows (Passer domesticus; 100 mg/kg = high, 60 mg/kg = medium, 30 mg/kg = low) and one-dose groups of zebra finches (Taeniopygia guttata; 3 mg/kg) and king quails (Coturnix chinensis; 26 mg/kg). Aerobic metabolism was measured during 1 h of exposure to subfreezing thermal conditions in low-dose house sparrows and king quails (26 mg/kg). Fenitrothion had no effect on metabolic rate during cold exposure or on blood hemoglobin at any time. By contrast, aerobic performance during exercise in sparrows was reduced by 58% (high), 18% (medium), and 20% (low), respectively, 2 d postdose. House sparrows (high) had the longest recovery period for peak metabolic rate (21 d) and plasma cholinesterase activity (14 d). House sparrows (high) and treated king quails had significantly lower myoglobin at 48 h postdose, whereas myoglobin was invariant in zebra finches and house sparrows (medium and low). Cholinesterase was maximally inhibited at 6 h postdose, and had recovered within 24 h, in house sparrows (low), king quails, and zebra finches. Exercise peak metabolic rate in zebra finches and king quails was reduced by 23% at 2 d and 3 d, respectively, despite these birds being asymptomatic in both behavior and plasma cholinesterase activities.

Fenitrothion, an organophosphate, affects running endurance but not aerobic capacity in fat-tailed dunnarts (Sminthopsis crassicaudata).

We measured aerobic metabolism during cold exposure and exercise performance (run duration and oxygen consumption while running at 1 m s(-1)) in the fat-tailed dunnart Sminthopsis crassicaudata, a dasyurid marsupial, before and after ingestion of 30 mg kg(-1) of fenitrothion, an organophosphate (OP) pesticide. Running endurance of OP-exposed animals was less than half that of control animals over the first 3 days after dosing and 55% of control animal endurance on day 5 post-dose. Despite these declines, peak metabolic rate at this running speed (9.3 times basal metabolic rate; BMR) was unaffected by OP exposure. Peak metabolic rate (PMR) and cumulative oxygen consumption during a 1-h exposure to conditions equivalent to -20 degrees C did not differ between OP-treated and control dunnarts, with PMR averaging 11 times BMR. We conclude that fenitrothion-induced exercise fatigue is not due to limitations in oxygen or substrate delivery to muscle or in their uptake per se, but more likely relates to decreased ability to sustain high-frequency neuromuscular function. The persistence of locomotor impairment following OP exposure in otherwise asymptomatic animals emphasizes the importance of using performance-based measures when characterising sublethal effects of pesticide exposure in an ecological context.

Increased probiotic yogurt or resistant starch intake does not affect isoflavone bioavailability in subjects consuming a high soy diet.

OBJECTIVE: Probiotics and prebiotics that affect gut microflora balance and its associated enzyme activity may contribute to interindividual variation in isoflavone absorption after soy intake, possibly enhancing isoflavone bioavailability. This study examined the effects of the consumption of bioactive yogurt (a probiotic) or resistant starch (a known prebiotic) in combination with high soy intake on soy isoflavone bioavailability. METHODS: Using a crossover design, chronic soy consumption was compared with soy plus probiotic yogurt or resistant starch in older male and postmenopausal females (n = 31). Isoflavone bioavailability was assessed at the beginning and end of each 5-wk dietary period by sampling plasma and urine after a standardized soy meal. RESULTS: Chronic soy intake did not significantly affect plasma or urinary isoflavones after the soy meal and there were no significant effects of probiotic or resistant starch treatment. However, there were trends for increased circulating plasma daidzein and genistein after the probiotic treatment and for increased plasma daidzein and genistein 24 h after soy intake with resistant starch treatment. Neither treatment induced or increased equol production, although there was a trend for increased plasma equol in "equol-positive" subjects (n = 12) after probiotic treatment. CONCLUSION: The weak or absence of effects of probiotic yogurt or resistant starch supplement to a chronic soy diet suggests that gut microflora were not modified in a manner that significantly affected isoflavone bioavailability or metabolism.

Cholinesterase response in native birds exposed to fenitrothion during locust control operations in eastern Australia.

Huge aggregations of flightless locust nymphs pose a serious threat to agriculture when they reach plague proportions but provide a very visible and nutritious resource for native birds. Locust outbreaks occur in spring and summer months in semiarid regions of Australia. Fenitrothion, an organophosphate pesticide, is sprayed aerially to control locust plagues. To evaluate fenitrothion exposure in birds attending locust outbreaks, we measured total plasma cholinesterase (ChE), butrylcholinesterase (BChE), and acetylcholinesterase (AChE) activities in four avian species captured pre- and postfenitrothion application and ChE reactivation in birds caught postspray only. Eleven of 21 plasma samples from four species had ChE activity below the diagnostic threshold (two standard deviations below the mean ChE activity of prespray samples). Granivorous zebra finches (Taeniopygia guttata) and insectivorous white-winged trillers (Lalage sueurii) had significantly lower mean plasma total ChE, BChE, and AChE activity postspray, while other insectivores, white-browed (Artamus superciliosus) and masked woodswallows (Artamus personatus), did not. Cholinesterase was reactivated in 19 of the 73 plasma samples and in one of three brain samples. We conclude that native bird species are exposed to fenitrothion during locust control operations. This exposure could have detrimental impacts, as both locust outbreaks and avian reproductive events are stimulated by heavy summer rainfall, leading to co-occurrence of locust control and avian breeding activities.

Fenitrothion, an organophosphorous insecticide, impairs locomotory function and alters body temperatures in Sminthopsis macroura (Gould 1845) without reducing metabolic rates during running endurance and thermogenic performance tests.

Endemic Australian mammal species are exposed to pesticides used for locust control as they occupy the same habitat as the target insect. The authors examined the impact of an ultra-low volume formulation of the organophosphorous insecticide fenitrothion (O,O-dimethyl-O-[3-methyl-4-nitrophenol]-phosphorothioate) on a suite of physiological measures that affect the ability of animals to survive in free-living conditions: locomotory and thermogenic functions, metabolic performance, body mass, and hematocrit and hemoglobin levels. Plasma and brain cholinesterase activity in relation to time since exposure to pesticide were also determined. An orally applied dose of 90 mg kg(-1) fenitrothion reduced running endurance in the stripe-faced dunnart, Sminthopsis macroura, by 80% the day after exposure concomitantly with a reduction of approximately 50% in plasma and 45% in brain acetylcholinesterase activity. These adverse effects disappeared by 10 d postexposure. Maximal metabolic rates reached during running were unaffected by pesticide, as were body mass and hemoglobin and hematocrit levels. Maximal cold-induced metabolic rate (measured as peak 2 min metabolic rate attained during cold exposure), time taken to reach peak metabolic rate on cold exposure, cumulative total oxygen consumed during shivering thermogenesis, and body temperature before and after cold exposure were unaffected by fenitrothion. Dunnart rectal temperatures showed a reduction of up to 5 °C after exposure to fenitrothion but returned to pre-exposure levels by 10 d postdose. Such physiological compromises in otherwise asymptomatic animals demonstrate the importance of considering performance-based measures in pesticide risk assessments.

A 'slow pace of life' in Australian old-endemic passerine birds is not accompanied by low basal metabolic rates.

Life history theory suggests that species experiencing high extrinsic mortality rates allocate more resources toward reproduction relative to self-maintenance and reach maturity earlier ('fast pace of life') than those having greater life expectancy and reproducing at a lower rate ('slow pace of life'). Among birds, many studies have shown that tropical species have a slower pace of life than temperate-breeding species. The pace of life has been hypothesized to affect metabolism and, as predicted, tropical birds have lower basal metabolic rates (BMR) than temperate-breeding birds. However, many temperate-breeding Australian passerines belong to lineages that evolved in Australia and share 'slow' life-history traits that are typical of tropical birds. We obtained BMR from 30 of these 'old-endemics' and ten sympatric species of more recently arrived passerine lineages (derived from Afro-Asian origins or introduced by Europeans) with 'faster' life histories. The BMR of 'slow' temperate-breeding old-endemics was indistinguishable from that of new-arrivals and was not lower than the BMR of 'fast' temperate-breeding non-Australian passerines. Old-endemics had substantially smaller clutches and longer maximal life spans in the wild than new arrivals, but neither clutch size nor maximum life span was correlated with BMR. Our results suggest that low BMR in tropical birds is not functionally linked to their 'slow pace of life' and instead may be a consequence of differences in annual thermal conditions experienced by tropical versus temperate species.

Acute oral toxicity of the organophosphorus pesticide fenitrothion to fat-tailed and stripe-faced dunnarts and its relevance for pesticide risk assessments in Australia.

The scarcity of information on the effects of pesticides on native Australian vertebrates constrains the development of biologically relevant risk assessments in Australia for the registration of pesticides. The concern that endemically old and unique Australian vertebrate fauna might display high sensitivity to pesticides used for locust control provoked examination of the acute oral toxicity of the organophosphorus pesticide fenitrothion for the fat-tailed dunnart, Sminthopsis crassicaudata (Gould 1844), and the stripe-faced dunnart, S. macroura (Gould 1845). By using the up-and-down method for determining acute oral toxicity, S. crassicaudata and S. macroura were found to have estimated median lethal doses (LD50s) of 129 mg/kg (95% confidence interval [CI] = 74.2-159.0) and 97 mg/kg (95% CI = 88.3-120.0), respectively. These values are 10 to 14 times lower than the reported LD50 values for a similar-sized eutherian mammal, Mus musculus (L. 1758; LD50 = 1,100-1,400 mg/kg) and lower than all other reported mammalian LD50 values. Such wide interspecific variation in sensitivity to fenitrothion may be a consequence of underlying differences in the metabolic pathway for fenitrothion detoxification in mammals and a possible explanation for the increased toxicity of fenitrothion to dunnarts, compared with other mammals, is proposed. The unexpectedly high sensitivity of these Australian marsupials to fenitrothion emphasises the importance of adequately evaluating the risks of pesticides to endemic Australian fauna.

Fipronil toxicity in northern bobwhite quail Colinus virginianus: reduced feeding behaviour and sulfone metabolite formation.

Fipronil is a phenyl pyrazole insecticide registered for agricultural use in many countries. Avian exposure to fipronil occurs mainly by ingesting contaminated insects or seeds. There is little information regarding the toxicological effects of fipronil in avian species and even less research documenting avian behavioural responses to fipronil ingestion. We examined the effects of a single oral dose of fipronil in northern bobwhite quail, the most fipronil-sensitive species tested to date, in respect to signs of intoxication and the metabolic fate of fipronil. Fipronil-treated birds did not eat or drink following pesticide administration, and as a result lost a significant amount of body mass. Treated birds also appeared withdrawn and did not respond to disturbance within the first hour after treatment. Identifiable signs of fipronil toxicity were not observed until at least 2d after treatment. Chemical analyses indicated a difference between fipronil and fipronil-sulfone residue distribution and bioaccumulation, with significantly higher (30- to 1000-fold) tissue concentrations of the sulfone detected at all time points from 8 to 96 h post-dose in brain, liver and adipose tissues. Tissue sulfone concentrations increased significantly in fipronil-treated birds, peaking at 72 h post-dose. Body mass decreased at all time points in dosed birds. The coincidence of the particular intoxication symptoms with the time course of rise in brain sulfone levels after fipronil dosing gives insight into possible mechanisms of toxicity in this highly sensitive species.

Frequency-dependent physiological trade-offs between competing colour morphs.

Evolutionary theory suggests that alternative colour morphs (i.e. genetically controlled phenotypes) may derive similar fitness under frequency-dependent selection. Here we experimentally demonstrate opposing effects of frequency-dependent social environments on plasma hormone levels (testosterone and corticosterone) and immune function between red- and black-headed male morphs of the Gouldian finch (Erythrura gouldiae). Red-headed males are highly sensitive to changes in the social environment, especially towards the relative density of their own aggressive morph, exhibiting high stress responses and immunosuppression in socially competitive environments. In contrast, the non-aggressive black-headed males follow a more passive strategy that appears to buffer them against social stresses. The differential effect of hormones on aggressive behaviour and immune performance reinforces the contrasting behavioural strategies employed by these colour morphs, and highlights the importance of the social environment in determining the individual basis of behavioural and physiological responses.

Steroid hormone interrelationships with territorial aggression in an Arctic-breeding songbird, Gambel's white-crowned sparrow, Zonotrichia leucophrys gambelii.

The breeding season is very brief for arctic-breeding passerines, and any interruptions of parental care by aggressive interactions over territory may reduce reproductive success. We tested both the "testosterone insensitivity" and "corticosterone insensitivity" hypotheses in the arctic-breeding Gambel's white-crowned sparrow, Zonotrichia leucophrys gambelii. Additionally, we tested whether simulated territorial intrusions (STIs), known to stimulate increases in luteinizing hormone (LH) and testosterone (T) in mid-latitude breeding Z. l. pugetensis, would also be effective in either the early or late phases of the brief breeding season of Z. l. gambelii. Plasma levels of T and LH were high early in the breeding season and declined as egg laying began. Exposure of free-living males to 10 min of STI significantly increased LH but not T secretion. Nonetheless, the pituitary-gonadal axis is sensitive as jugular injection of gonadotrophin-releasing hormone increased plasma T at 10 min relative to saline-challenged controls. T implants failed to increase territorial aggression following STI during incubation. These data are consistent with the T insensitivity hypothesis and contrast sharply with the response of the southerly breeding subspecies, Z. l. pugetensis, in which the territorial response to T administration is retained throughout its relatively long breeding season. However, corticosterone implants during the incubation period decreased territorial aggression during STI. This responsiveness to corticosterone is not consistent with the corticosterone insensitivity hypothesis of stress modulation. Z. l. gambelii retain sensitivity to corticosterone levels that may occur naturally in response to environmental perturbations resulting in suppression of territorial behavior.

Limited lipid-lowering effects of regular consumption of whole soybean foods.

AIM: To examine cardiovascular health benefits of foods containing a whole soybean extract. METHODS: The study design was a randomized, placebo-controlled crossover trial of consuming soy-based milk and yoghurt (treatment) or equivalent dairy products (control) for 5 weeks each. Twenty-six mildly hypercholesterolaemic and/or hypertensive volunteers were recruited from the community as study volunteers, of which 23 completed. Main outcome measures included clinic and ambulatory blood pressure, arterial compliance, lipids, fatty acids and isoflavones in fasted blood and 24-hour urinary isoflavone excretion. Nutrient intakes were assessed initially and after each 5-week period. Multiple regression analyses were used to determine predictor variables in statistical models; order effects were tested by repeated measures ANOVA. Changes in Lp(a) were determined by Wilcoxon signed ranks tests; other differences between treatment and control were assessed by t tests. RESULTS: Plasma and urinary isoflavones were markedly increased by whole soy supplementation but there were no overall differences in plasma lipids, blood pressure or arterial compliance between the soy and dairy diets. However, in 8 equol-positive subjects (equol detected in either plasma or urine), retrospective analysis revealed significant reductions in total cholesterol (8.5%), LDL cholesterol (10%), LDL:HDL ratio (13.5%), plasma triglycerides (21%) and lipoprotein(a) (11%) with the soy diet. These reductions were independent of changes in polyunsaturated fat and other macronutrient intakes. CONCLUSIONS: Regular consumption of whole soybean milk and yogurt products had no effect on plasma lipids, blood pressure or arterial compliance in at-risk subjects, despite substantially increasing isoflavone levels in blood and urine. Retrospective analysis suggests that improvement of plasma lipids may have been limited to equol-positive subjects.