Molecular and immunological analysis of hen's egg yolk allergens with a focus on YGP42 (Gal d 6)

Allergy to hen's (Gallus domesticus) egg white is one of the most common forms of food allergy. Allergy to hen's yolk also exists however, to a lesser extent when compared to egg white allergy. Two minor allergens from the hen's egg yolk known as α-livetin (Gal d 5) and YGP42 (Gal d 6) were discovered recently. In this study, we investigated whether sensitization to egg white is associated with reactivity to egg yolk as well. Sera obtained from 25 patients with allergy to egg white were tested for specific IgE binding for egg yolk proteins through western immunoblotting. 36% of patients were found with true IgE-sensitization against egg yolk proteins. It was found that most of the IgE reactive yolk proteins were fragments of major precursor proteins of hen; vitellogenin-1 (VTG-1), vitellogenin-2 (VTG-2) and apolipoprotein B (Apo B). The egg yolk allergen Gal d 6 is the C-terminal part of VTG-1 and was found to be allergenic in significant percentage of egg white allergy patients. These results highlight the significance of Gal d 6 as an important allergen of egg yolk. Therefore, the secondary aim of this study involved developing a recombinant version of YGP42 in an Escherichia coli expression system. Recombinant Gal d 6 (rGal d6) was expressed as a fusion peptide with a 6 × His tag and purified using metal chelating resin. The inhibition ELISA results showed that rYGP42 was IgE reactive and was able to inhibit IgE binding to crude egg yolk (CEY) by up to 30%. Traditionally, it was thought that allergy to egg yolk occurred independently from sensitization to egg white. This study underlies the importance of concomitant sensitization to egg yolk proteins in patients allergic to egg white. Evidence reported in this study strongly suggests that egg yolk has potentially undiscovered allergens and therefore warrants further investigation. Furthermore, IgE reactive Gal d 6 presented in this study has the potential to be used in diagnosis and immunotherapy to treat egg allergy.

Testosterone, dominance signalling and immunosuppression in the house sparrow, Passer domesticus

The immunocompetence handicap hypothesis (ICHH) suggests that dominance signals are costly because their development is controlled by testosterone, which is immunosuppressive. Signal control therefore links an increased disease risk with a high quality signal. The chest bib of the house sparrow, Passer domesticus, is a signal known to be related to dominance and under control of testosterone levels. We experimentally manipulated testosterone in male sparrows during the breeding season and again independently during the post-breeding period to test whether variation in levels of testosterone could cause variation in levels of immunocompetence. There was no effect of testosterone manipulation on the cell-mediated response of birds to phytohaemagglutinin injection, nor did testosterone levels appear to affect either white blood cell ratios or red blood cell counts. In contrast, both breeding season and post-breeding season testosterone levels had significant effects upon the humoral response of the birds to sheep red blood cell injections. However, whilst testosterone during the breeding season appeared to act immunosuppressively, the role of post-breeding levels is less clear. In concordance with a previous study, there was an indication that corticosterone is involved in mediating the immunosuppressive effects of testosterone. The strength of the secondary humoral response and the cell-mediated response were negatively related suggesting the possibility of a trade-off between the different arms of the immune system. These results provide some support for the ICHH as a mechanism promoting the evolution of costly badges of status, although the results question whether the immunosuppressive cost can be mediated by testosterone at the time of badge development.

Does testosterone determine dominance in the house sparrow Passer domesticus? An experimental test

Badges of status function in many birds within a social context to establish dominance hierarchies and reduce antagonistic encounters. In order to maintain the honesty of the signalling system, such badges must be costly to produce or to maintain. The chest bib of the house sparrow functions as a badge of status and changes in size are known to be controlled by testosterone levels. We sought to test the relative importance of testosterone as opposed to bib size in determining dominance within a group of male house sparrows. We did this by manipulating testosterone levels independently during both breeding and post-breeding seasons in experimental birds and examining the effect of testosterone titre, as well as corticosterone titre relative to bib size on dominance levels. Dominance hierarchies within the groups were tested during both the breeding and post-breeding phases. We compared the results of these tests with dominance among intact (unmanipulated) birds. Results suggested that the breeding season dominance levels were largely determined by testosterone levels as well as bib size, whereas the post-breeding dominance levels were determined by postbreeding testosterone titre and previous breeding season dominance level. Within unmanipulated birds, basal corticosterone levels were significantly, negatively correlated with dominance level, but only during the breeding season. The influence of breeding season dominance on post-breeding dominance suggests social history is important in determining dominance interactions as well as current testosterone levels and bib size.

The role of testosterone in bib size determination in the male house sparrow Passer domesticus, is age dependent

Secondary sexual signals are thought to indicate individual quality. In order to understand the evolutionary pressures that give rise to such traits it is important to understand the physiological mechanisms underlying their production. The black bib of the house sparrow Passer domesticus is known to function as a badge of social status in males. Past studies have found that the size of the bib in older males is determined, at least partly, by the androgen testosterone. The immunocompetence handicap hypothesis suggests that testosterone has a key role in maintaining honest signalling – it is both involved in the development or expression of sexual signals and is immunosuppressive. In this paper we test experimentally two hypo theses relating to bib size development, whether 1) testosterone is only immunosuppressive in conditions where the natural feedback loop from the testes has been removed, and 2) testosterone is, in addition to influencing the bib size of older males, responsible for the size of the bib in juvenile sparrows. In the first experiment we found that exogenous testosterone administered to intact males during the winter (when LH and FSH levels are very low and were not artificially increased by castration) caused significant immunosuppression, albeit in interaction with the stress hormone corticosterone. Second, we found that exogenous testosterone administration in castrated fledgling male house sparrows had no effect on subsequent post-juvenile moult bib size relative to controls. Our results suggest that in some circumstances testosterone can be immunosuppressive, but that its role in bib size determination is age-dependent.

The power of myth : the (non) impact of invasive cane toads (Bufo marinus) on domestic chickens (Gallus gallus)

Myths about invasive species are widespread in the general community, even when contrary to scientific evidence. Public revulsion against invasive cane toads (Bufo marinus) in Australia has encouraged the belief that toads pose a significant risk to domestic poultry, by poisoning fowls that eat toads or that drink water contaminated by toads. Although discredited by scientists in 1938, within 2 years of the toads’ introduction to Australia, the myth continues to flourish. We conducted experimental trials to evaluate the vulnerability of chickens to toad-contaminated water, and to toad ingestion. No ill effects were seen, with one chicken consuming 45 small toads without falling ill. Thus, available evidence suggests that cane toads do not imperil domestic poultry.

An experimental examination of interindividual variation in feather corticosterone content in the House Sparrow, Passer domesticus in southeast Australia

Non-invasive techniques for measuring glucocorticoids (GCs) have become more prevalent, due to the advantage of eliminating the effects of animal disturbance on GC levels and their potential to provide an integrated, historic estimate of circulating GC levels. In the case of birds, corticosterone (CORT) is deposited in feathers, and may reflect a bird's GC status over the period of feather synthesis. This technique thus permits a retrospective view of the average circulating GC levels during the moult period. While it is generally assumed that differences in feather CORT content (CORTf) between individuals reflects their different stress histories during either natural or induced moult, it is not clear how much of this variation is due to extrinsic versus intrinsic factors. We examined this question by determining CORTf in free-living house sparrows (Passer domesticus) from two populations, one urban and the other rural, that were plucked before and after exposure to different plasma CORT levels while held captive. We experimentally manipulated plasma CORT by implanting birds with either a corticosterone-filled, metyrapone-filled, or empty ('sham') silastic capsule as replacement feathers first emerged. The pattern of post-treatment CORTf was consistent with our expectations, based on plasma CORT levels of an experimentally implanted reference group. However, there was no statistically significant difference in CORTf between these treatment groups unless sex, population origin, and CORTf of original feathers for each individual were included in a model. Thus, birds with higher CORTf in feathers removed for this experiment tended to have higher CORTf in post-treatment replacement feathers, irrespective of treatment. In addition, we found that feather fault bar scores were significantly higher in CORT-treated birds than in the other two treatment groups, but did not vary directly with CORTf level. Our study therefore broadly confirms the use of feathers as a non-invasive tool to estimate plasma CORT during moult in birds, but importantly demonstrates the potential for intrinsic differences in stress characteristics between populations and individuals to obscure the effects extrinsic stressors might have on CORTf .

Effects of isolation on stress responses to novel stimuli in Subadult chickens (Gallus gallus)

Extensive research has examined the effects of social isolation in neonatal and adult animal populations, but few studies have examined the effect of social isolation in early adulthood. Animals reaching reproductive age often experience extensive social changes as they leave their natal site, and a social stressor like isolation may uniquely affect this age group. Furthermore, adolescence is a time when sex differences in behavior become more pronounced. As such, the effects of social stressors are likely to vary by sex. In this study, we used noninvasive methods to evaluate stress responses to social change in male and female subadult chickens (Gallus gallus). Half of the birds experienced regular sessions of social isolation over the course of 2 wk, while the other half were never isolated. Subsequently, all of the animals were exposed to a suite of three novel probes, including an open-field test. We monitored the birds’ behavioral (head movements) and physiological (fecal glucocorticoid metabolites, FGM) response to the tests. Our results indicate that, for subadult chickens, the effect of social isolation is sex dependent: Male FGM and behavioral responses did not change with subsequent experiences, in contrast to females. Females also exhibited more social reinstatement behavior compared to males. Our results are consistent with the expectations of differences between the sexes based on changes in the social environment due to sex-biased dispersal patterns. For both sexes, the FGM and behavioral responses varied independently, which highlights the necessity for multiple measures of stress in animal populations.

Powerline corridors: degraded ecosystems or wildlife havens?

Management of powerline corridors in Australia has traditionally focused on the complete removal of vegetation using short rotation times owing to the perceived hazard of fire associated with corridor vegetation. Because of the intense management associated with fire hazards, little thought has been given to use of powerline corridors by wildlife. This has resulted in corridors traditionally being viewed as a source of fragmentation and habitat loss within forested ecosystems. We investigated the responses of small mammal communities living in a powerline corridor to management-induced vegetation changes at different successional stages, to determine whether a compromise could be reached between managing corridors for fire and biodiversity. Habitat modelling in the corridor and adjacent forest for three native and one introduced small mammal species demonstrated that species responded to changes in vegetation structural complexity, rather than time-since-management per se. Early seral stages of vegetation recovery after corridor management encouraged the introduced house mouse (Mus domesticus) into corridors and contributed little to biodiversity. Mid-seral-stage vegetation, however, provided habitat for native species that were rare in adjacent forest habitats. As the structural complexity of the vegetation increased, the small mammal community became similar to that of the forest so that corridor vegetation contributed fewer biodiversity benefits while posing an unacceptable fire risk. If ecologically sensitive management regimes are implemented to encourage mid-seral vegetation and avoid complete vegetation removal, powerline corridors have the potential to improve biodiversity. This would maintain landscape connectivity and provide habitat for native species uncommon in the forest while still limiting fuel loads in the corridor.

The diet of a Southern Boobook Ninox novaeseelandiae in Box-ironbark country, central Victoria

The diet of a Southern Boobook Ninox novaeseelandiae in a Box-Ironbark woodland remnant in central Victoria was studied. An analysis of 42.5 pellets found invertebrates to represent 82% of the total prey items, but vertebrates, in the form of the House Mouse Mus domesticus, to represent 88% of the biomass. The proportion of spiders as prey items in the diet in this instance (43%) was significantly higher than in other studies on mainland Australia.

Occurrence of small mammals in a fragmented landscape: the role of vegetation heterogeneity

Most studies of habitat use by small mammals rely on data from trapping grids. Such studies pertain to habitat use by individuals, which may not reflect population-level requirements. To meet the challenge of landscape change, it is important to understand habitat use by populations across large geographic areas. We surveyed small mammals in 48 forest remnants across a 300 km2 study area, to investigate the influence of vegetation heterogeneity on regional distributions. Information-theoretic techniques were used to evaluate models of vegetation associations. Richness of native mammals was influenced by vegetation condition: disturbed sites supported fewer species. Models for individual species showed the agile antechinus, Antechinus agilis, to prefer structurally diverse forest vegetation, the long-nosed potoroo, Potorous tridactylus, to favour mesic shrub communities, the bush rat, Rattus fuscipes, to prefer complex low cover regardless of composition, the swamp rat, Rattus lutreolus, to favour reduced canopy cover, and the house mouse, Mus domesticus, to prefer disturbed vegetation. To satisfy the needs of all native species, a mosaic of natural vegetation is required. Degradation and simplification of forest vegetation have detrimental consequences. These results highlight the need to consider habitat quality, together with more traditional biogeographic variables, when investigating factors influencing patch occupancy by native fauna in modified landscapes.

Diet of an eastern barn owl Tyto javanica on the Patho Plains, northern Victoria

A small sample of eastern Barn Owl Tyto javanica pellets, from native grasslands on the Patho Plains in northern Victoria in February 2007, contained the remains of 48 prey individuals: 38 Australian Plague Locusts Chortoicetes terminifera, nine house Mice Mus domesticus and one Fat-tailed dunnart Sminthopsis crassicaudata. Such a high proportion of locusts in the eastern Barn Owl's diet is noteworthy, and is discussed in the context of recent locust-spraying operations in the region.