Andrew T. D. Bennett

Infectious diseases can drive populations and species to extinction. Beak and feather disease virus (BFDV) is a circovirus of global conservation concern that can infect all Psittaciformes and some other species. Yet some parrot species, such as Crimson rosellas (Platycercus elegans), can live successfully with high BFDV prevalence (>40%) with no clinical signs reported in infected individuals. We assessed BFDV load in 10-12 tissues per bird, from n = 66 P. elegans, to reveal tissue tropism and BFDV persistence in tissues. Here we show that in 94% of individuals, BFDV was detected in one or more tissues. While BFDV replicated to high levels in subadults, in adults (some confirmed seropositive) the virus persisted in various tissues at much lower levels. Our findings reveal that BFDV is much more common in wild P. elegans than previously thought and suggest that current screening practices (mostly on blood) may substantially underestimate BFDV infection estimates, with implications for biosecurity and conservation programs globally.

Nature provides a rich source of information for the design of novel materials; yet there remain significant challenges in the design and manufacture of materials that replicate the form, function, and sustainability of biological solutions. Here, we identify key challenges and promising approaches to the development of materials informed by biology. These challenges fall into two main areas; the first relates to harnessing biological information for materials innovation, including key differences between biological and synthetic materials, and the relationship between structure and function. We propose an approach to materials innovation that capitalizes on biodiversity, together with high-throughput characterization of biological material architectures and properties, linked to environmental and ecological context. The second area relates to the design and manufacture of bioinformed materials, including the physical scale of material architectures and manufacturing scale up. We suggest ways to address these challenges and promising prospects for a bioinformed approach to materials innovation.

Background: Beak and feather disease virus (BFDV) is a circovirus that infects captive and wild psittacine birds, and is of conservation concern. The haemagglutination inhibition (HI) assay is used to determine antibody titres against BFDV, and the use of dried blood spots (DBS) on filter paper stored at room temperature has been suggested to be an equally valid technique to the use of frozen serum. However, research on other pathogens has found variable results when investigating the longevity of antibodies stored on DBS at room temperature. Consequently, we aimed to test the temporal stability of antibodies to BFDV in DBS samples stored long-term at room temperature. A further goal was to add to the current knowledge of antibody response to naturally acquired BFDV infection in crimson rosellas (Platycercus elegans).Methods: Blood was collected from wildP. elegansin Victoria, Australia, that had been live-trapped (n= 9) or necropsied (n= 11). BFDV virus load data were obtained from blood stored in ethanol by real-time quantitative PCR (qPCR); antibody titres were obtained by HI assay from either DBS or serum samples, which had been collected concurrently. All HI assays were performed commercially by the Veterinary Diagnostic Laboratory (VDL) in Charles Sturt University, Australia, who were blind to BFDV blood status.Results: HI titres from DBS stored at room temperature declined significantly over time (~80 weeks). By contrast, frozen serum samples assayed after 80 weeks in storage all had high HI titres, only varying up to one dilution step from the initial HI titres obtained from DBS at 3–6 weeks after sampling. Weak HI titres from DBS samples all came back negative when the test was repeated only nine weeks later. Novel high HI titres were reported inP. elegans, and while most birds with high antibody titres had corresponding negative qPCR results, a single subadult presented with high HI titres and virus load simultaneously.Conclusion: Detection of antibodies on filter paper stored at room temperature decreases over time, increasing the chances of false negatives in these samples, and in repeated testing of samples with weak HI titres. Consequently, serum should be the preferred sample type to use for seroepidemiological studies on BFDV in parrots and other bird species. When not possible, it may help to store DBS on filter paper at −20 °C or lower. However, prompt testing of DBS samples (e.g., P. elegans, especially adults, can produce high antibody titres against BFDV, which may help them resist infection.

Wildlife can be hosts for pathogens of conservation and zoonotic concern. Infection with one pathogen can increase the risk of co-infection with others, with resulting impacts on hosts, for example on body condition or survival. However, studies of co-infection in wild populations are rare. Globally, psittacine birds can be infected with beak and feather disease virus (BFDV), a circovirus of significant conservation concern, and with bacteria of the order Chlamydiales, including the zoonotic Chlamydia psittaci. These pathogens are common in Australian psittacines, and their putatively immunosuppressive effects would predict co-infection to be more common than expected due to chance alone. Here, we test this hypothesis across four abundant and widespread Australian parrot species: Crimson Rosella, Sulphur-crested Cockatoo, Galah and Blue-winged Parrot. Using wild-caught birds, we tested for pathogen DNA and, in the case of Chlamydiales, also antibodies, to investigate both current and previous infection status. We detected co-infection prevalence of up to 16.4% in the combined samples from the four species and up to 28.9% in Crimson Rosellas, the species with the largest sample size. Contrary to expectations, however, we found that the incidence of infections with the two pathogens was statistically independent. There was also no effect of co-infection on body condition or haematocrit. Our findings therefore suggest that while concurrent infection does occur, it is no more common than expected by chance. We discuss limitations of the data and suggest avenues for future research. Overall, our study provides insights that potentially benefit psittacine conservation, welfare, and human health.

The Chlamydia are a globally distributed genus of bacteria that can infect and cause disease in a range of hosts. Birds are the primary host for multiple chlamydial species. The most well-known of these is Chlamydia psittaci, a zoonotic bacterium that has been identified in a range of wild and domesticated birds. Wild birds are often proposed as a reservoir of Chlamydia psittaci and potentially other chlamydial species. The aim of this review is to present the current knowledge of chlamydial infections in wild avian populations. We focus on C. psittaci but also consider other Chlamydiaceae and Chlamydia-related bacteria that have been identified in wild birds. We summarise the diversity, host range, and clinical signs of infection in wild birds and consider the potential implications of these infections for zoonotic transmission and avian conservation. Chlamydial bacteria have been found in more than 70 species of wild birds, with the greatest chlamydial diversity identified in Europe. The Corvidae and Accipitridae families are emerging as significant chlamydial hosts, in addition to established wild hosts such as the Columbidae. Clarifying the effects of these bacteria on avian host fitness and the zoonotic potential of emerging Chlamydiales will help us to understand the implications of these infections for avian and human health.

Chlamydia psittaci (order: Chlamydiales) is a globally distributed zoonotic bacterium that can cause potentially fatal disease in birds and humans. Parrots are a major host, yet prevalence and risk factors for infection in wild parrots are largely unknown. Additionally, recent research suggests there is a diverse range of novel Chlamydiales circulating in wildlife. We therefore sampled seven abundant parrot species in south-eastern Australia, taking cloacal swabs and serum from n = 132 wild adults. We determined C. psittaci and Chlamydiales prevalence and seroprevalence, and tested for host species, sex, geographical and seasonal differences, and temporal changes in individual infection status. Across all species, Chlamydiales prevalence was 39.8% (95% CI 31.6, 48.7), C. psittaci prevalence was 9.8% (95% CI 5.7, 16.3) and C. gallinacea prevalence was 0.8% (95% CI 0.1, 4.5). Other Chlamydiales species were not identified to species level. We identified two C. psittaci strains within the 6BC clade, which is highly virulent in humans. Seroprevalence was 37.0% (95% CI 28.5, 46.4). Host species (including crimson rosellas, galahs, sulphur-crested cockatoos and blue-winged parrots) differed in seroprevalence and Chlamydiales prevalence. Galahs had both highest Chlamydiales prevalence (54.8%) and seroprevalence (74.1%). Seroprevalence differed between sites, with a larger difference in males (range 20–63%) than females (29–44%). We reveal a higher chlamydial prevalence than previously reported in many wild parrots, with implications for potential reservoirs, and transmission risks to humans and other avian hosts.

Pathogens pose a major risk to wild host populations, especially in the face of ongoing biodiversity declines. Beak and feather disease virus (BFDV) can affect most if not all members of one of the largest and most threatened bird orders world-wide, the Psittaciformes. Signs of disease can be severe and mortality rates high. Its broad host range makes it a risk to threatened species in particular, because infection can occur via spill-over from abundant hosts. Despite these risks, surveillance of BFDV in locally abundant wild host species has been lacking. We used qPCR and haemagglutination assays to investigate BFDV prevalence, load and shedding in seven abundant host species in the wild in south-east Australia: Crimson Rosellas (Platycercus elegans), Eastern Rosellas (Platycercus eximius), Galahs (Eolophus roseicapillus), Sulphur-crested Cockatoos (Cacatua galerita), Blue-winged Parrots (Neophema chrysostoma), Rainbow Lorikeets (Trichoglossus moluccanus) and Red-rumped Parrots (Psephotus haematonotus). We found BFDV infection in clinically normal birds in six of the seven species sampled. We focused our analysis on the four most commonly caught species, namely Crimson Rosellas (BFDV prevalence in blood samples: 41.8%), Sulphur-crested Cockatoos (20.0%), Blue-winged Parrots (11.8%) and Galahs (8.8%). Species, but not sex, was a significant predictor for BFDV prevalence and load. 56.1% of BFDV positive individuals were excreting BFDV antigen into their feathers, indicative of active viral replication with shedding. Being BFDV positive in blood samples predicted shedding in Crimson Rosellas. Our study confirms that BFDV is endemic in our study region, and can inform targeted disease management by providing comparative data on interspecies variation in virus prevalence, load and shedding.

Members of the Chlamydia genus are known to cause disease in both humans and animals. A variety of other species in the order Chlamydiales are increasingly being discovered and emerging as potential pathogens, yet there are scarce data on the diversity, prevalence and impacts of these pathogens in wild birds. To address this gap, we investigated which Chlamydiales species are present in a wild population of a common Australian parrot, the Crimson Rosella (Platycercus elegans) . We collected cloacal swabs and serum from 136 individuals in south eastern Australia, over two years, and tested several predictors of prevalence: age, sex, season, and breeding status. We used multiple PCR assays to determine bacterial prevalence in cloacal swabs and a solid‐phase ELISA to determine seroprevalence. We found Chlamydiales PCR prevalence of 27.7% (95% CI 20.2, 36.2), and identified at least two families (Chlamydiaceae and Parachlamydiaceae ). Regarding known chlamydial avian pathogens, we found C. psittaci at 6.2% (95% CI 2.7, 11.8) and C. gallinacea at 4.6% (95% CI 1.7, 9.8) prevalence. We also identified at least two potentially novel Chlamydiales species, of unknown pathogenicity. Sex and breeding status predicted Chlamydiales PCR prevalence, with females more likely to be infected than males, and non‐breeding birds more likely to be infected than breeding birds. Seroprevalence was 16% (95% CI 8.8, 25.9). Season and breeding status were strong predictors of seroprevalence, with highest seroprevalence in autumn and in non‐breeding birds. Our results reveal a diversity of Chlamydiales species in this abundant wild host, and indicate that host‐specific and temporal factors are associated with infection risk. Our findings suggest that wild parrots are a reservoir of both known and novel Chlamydiales lineages, of zoonotic and pathogenic potential.