A blurring of life-history lines: Immune function, molt and reproduction in a highly stable environment.

Rufous-collared sparrows (Zonotrichia capensis peruviensis) from valleys in the Atacama Desert of Chile, live in an extremely stable environment, and exhibit overlap in molt and reproduction, with valley-specific differences in the proportion of birds engaged in both. To better understand the mechanistic pathways underlying the timing of life-history transitions, we examined the relationships among baseline and stress-induced levels of corticosterone (CORT), testosterone, and bacteria-killing ability of the blood plasma (BKA), as well as haemosporidian parasite infections and the genetic structure of two groups of sparrows from separate valleys over the course of a year. Birds neither molting nor breeding had the lowest BKA, but there were no differences among the other three categories of molt-reproductive stage. BKA varied over the year, with birds in May/June exhibiting significantly lower levels of BKA than the rest of the year. We also documented differences in the direction of the relationship between CORT and BKA at different times during the year. The direction of these relationships coincides with some trends in molt and reproductive stage, but differs enough to indicate that these birds exhibit individual-level plasticity, or population-level variability, in coordinating hypothalamo-pituitary-adrenal axis activity with life-history stage. We found weak preliminary evidence for genetic differentiation between the two populations, but not enough to indicate genetic isolation. No birds were infected with haemosporidia, which may be indicative of reduced parasite pressure in deserts. The data suggest that these birds may not trade off among different life-history components, but rather are able to invest in multiple life-history components based on their condition.

Chronically raised glucocorticoids reduce innate immune function in Belding's ground squirrels (Urocitellus beldingi) after an immune challenge.

The hypothalamic-pituitary-adrenal (HPA) axis releases glucocorticoids (GCs), or stress hormones, during the vertebrate stress response. GCs can both enhance and suppress the immune system depending on whether the experienced stressor is acute or chronic and what aspect of immune function is measured. More research is needed to fully understand how the immune system reacts to stressors. In this study, we examined the effects of chronically raised GCs on innate immune function in Belding's ground squirrels (Urocitellus beldingi). We measured immune function with a bacteria killing ability (BKA) assay, an integrative and functional assessment of an animal's ability to clear a bacterial infection. All studies to date have examined how acute stressors or repeated social stressors impact BKA. This study is the first to our knowledge to investigate how chronically raised GCs impact BKA both before and after an immune challenge. We noninvasively raised GCs in treatment squirrels for six days and then gave them, and a group of untreated (control) squirrels, an injection of lipopolysaccharide (LPS) to stimulate their innate immune system. Treatment squirrels exhibited lower BKA after, but not before, being challenged with LPS. These results suggest that experiencing chronic stress may not be detrimental to immune functioning until an individual is challenged with an infection.

Egg Incubation Temperature Affects Development of Innate Immune Function in Nestling American Robins (Turdus migratorius).

The innate immune system provides important first-line defenses against invading pathogens and is considered especially important for developing organisms. However, we know little about how early-life conditions influence these defenses in wild animals. For oviparous species such as birds, embryonic development occurs in the egg, which can be subject to variation in thermal conditions. There is evidence from cavity-nesting species and species with precocial young that reduced incubation temperatures can result in reduced measures of innate immunity. Whether and how this thermal variation impacts innate immunity for open-cup-nesting species with altricial offspring has not been examined. In this study, we experimentally manipulated egg incubation temperature for American robins (Turdus migratorius) and compared the bacteria-killing ability (BKA) of the nestlings' blood plasma. We collected baseline and poststressor samples on day 7 and day 10 after hatch to gain additional insights into the ontogeny of this immune measure, as well as into whether any changes were linked to levels of the glucocorticoid hormone corticosterone (CORT). We found that nestlings incubated at the low treatment (36.1°C) had significantly reduced BKA compared with nestlings incubated at the high treatment (37.8°C) when controlling for the posthatch nest environment. We also documented a significant reduction in poststressor levels of BKA, as well as an increase in BKA from day 7 to day 10. We found a weak inverse association between CORT and BKA but no other indications that BKA was mediated via treatment-induced variation in CORT. Our results suggest that incubation temperature can affect development of innate immunity in open-cup-nesting passerines.

The Role of Properdin in Killing of Non-Pathogenic Leptospira biflexa.

Properdin (P) is a positive regulatory protein that stabilizes the C3 convertase and C5 convertase of the complement alternative pathway (AP). Several studies have suggested that properdin can bind directly to the surface of certain pathogens regardless of the presence of C3bBb. Saprophytic Leptospira are susceptible to complement-mediated killing, but the interaction of properdin with Leptospira spp. has not been evaluated so far. In this work, we demonstrate that properdin present in normal human serum, purified properdin, as well as properdin oligomers P2, P3, and P4, interact with Leptospira. Properdin can bind directly to the bacterial surface even in the absence of C3b. In line with our previous findings, AP activation was shown to be important for killing non-pathogenic L. biflexa, and properdin plays a key role in this process since this microorganism survives in P-depleted human serum and the addition of purified properdin to P-depleted human serum decreases the number of viable leptospires. A panel of pathogenic L.interrogans recombinant proteins was used to identify putative properdin targets. Lsa30, an outer membrane protein from L. interrogans, binds to unfractionated properdin and to a lesser extent to P2-P4 properdin oligomers. In conclusion, properdin plays an important role in limiting bacterial proliferation of non-pathogenic Leptospira species. Once bound to the leptospiral surface, this positive complement regulatory protein of the AP contributes to the formation of the C3 convertase on the leptospire surface even in the absence of prior addition of C3b.

Agricultural Intensification Is Linked to Constitutive Innate Immune Function in a Wild Bird Population.

Immunity represents an important defense mechanism against pathogens and is intimately linked to fitness. Previous studies have found significant interindividual variation of immune responses in wild populations and have emphasized the importance of ecological factors in explaining this variability. A deterioration of environmental conditions, such as agricultural intensification, can modify resource availability and, as a result, alter immune functions. The aim of this study was to investigate the effect of agricultural intensification on innate immune functions in breeding adult tree swallows (Tachycineta bicolor) over 6 yr. We investigated the relationship between agricultural intensification and bacteria-killing ability (BKA) against a strain of E. coli. We also assessed how the presence and the number of bacteria on the surface of eggshells changed with agricultural intensification. Contrary to our expectations, we found a positive relationship between agricultural intensification and BKA, where individuals had higher BKA in intensive compared to nonintensive farmlands. While this trend was observed through the 6-yr study, we also found a tendency for a reduction of BKA over time. We found no relationships between the presence or number of different types of eggshell bacteria and agricultural intensification or an association between BKA of females and eggshell bacteria of their clutch. Our results suggest that differences in immune function observed between intensive and nonintensive farmlands could reflect genetic differences among individuals and/or trade-offs with other traits expressed among habitats within our study system.

Correlates of immune defenses in golden eagle nestlings.

An individual's investment in constitutive immune defenses depends on both intrinsic and extrinsic factors. We examined how Leucocytozoon parasite presence, body condition (scaled mass), heterophil-to-lymphocyte (H:L) ratio, sex, and age affected immune defenses in golden eagle (Aquila chrysaetos) nestlings from three regions: California, Oregon, and Idaho. We quantified hemolytic-complement activity and bacterial killing ability, two measures of constitutive immunity. Body condition and age did not affect immune defenses. However, eagles with lower H:L ratios had lower complement activity, corroborating other findings that animals in better condition sometimes invest less in constitutive immunity. In addition, eagles with Leucocytozoon infections had higher concentrations of circulating complement proteins but not elevated opsonizing proteins for all microbes, and eagles from Oregon had significantly higher constitutive immunity than those from California or Idaho. We posit that Oregon eagles might have elevated immune defenses because they are exposed to more endoparasites than eagles from California or Idaho, and our results confirmed that the OR region has the highest rate of Leucocytozoon infections. Our study examined immune function in a free-living, long-lived raptor species, whereas most avian ecoimmunological research focuses on passerines. Thus, our research informs a broad perspective regarding the evolutionary and environmental pressures on immune function in birds.

The Role of properdin in killing of non-pathogenic Leptospira biflexa

Properdin (P) is a positive regulatory protein that stabilizes the C3 convertase and C5 convertase of the complement alternative pathway (AP). Several studies have suggested that properdin can bind directly to the surface of certain pathogens regardless of the presence of C3bBb. Saprophytic Leptospira are susceptible to complement-mediated killing, but the interaction of properdin with Leptospira spp. has not been evaluated so far. In this work, we demonstrate that properdin present in normal human serum, purified properdin, as well as properdin oligomers P2, P3, and P4, interact with Leptospira. Properdin can bind directly to the bacterial surface even in the absence of C3b. In line with our previous findings, AP activation was shown to be important for killing non-pathogenic L. biflexa, and properdin plays a key role in this process since this microorganism survives in P-depleted human serum and the addition of purified properdin to P-depleted human serum decreases the number of viable leptospires. A panel of pathogenic L. interrogans recombinant proteins was used to identify putative properdin targets. Lsa30, an outer membrane protein from L. interrogans, binds to unfractionated properdin and to a lesser extent to P2-P4 properdin oligomers. In conclusion, properdin plays an important role in limiting bacterial proliferation of non-pathogenic Leptospira species. Once bound to the leptospiral surface, this positive complement regulatory protein of the AP contributes to the formation of the C3 convertase on the leptospire surface even in the absence of prior addition of C3b.