A single Haemoproteus plataleae haplotype is widespread in white ibis (Eudocimus albus) from urban and rural sites in southern Florida.

The American white ibis (Eudocimus albus), a common bird species in Florida, has become increasingly urban, with many populations relying heavily on urban and suburban habitats, which may alter parasite transmission. Parasites of ibis, especially haemosporidians, are understudied. Avian haemosporidia can have a wide range of impacts on birds, including decreased reproductive success or increased mortality. Because southern Florida is subtropical and has a high diversity of potential vectors for haemosporidia, we hypothesized that there will be a high prevalence and genetic diversity of haemosporidia in white ibis. A total of 636 ibis from South Florida were sampled from 2010 to 2022, and blood samples were tested for haemosporidia by examination of Giemsa-stained thin blood smears and/or nested PCRs targeting the cytochrome b gene. A total of 400 (62.9%, 95% CI 59-66.7%) ibis were positive for parasites that were morphologically identified as Haemoproteus plataleae. Sequences of 302 positives revealed a single haplotype of Haemoproteus (EUDRUB01), which was previously reported from white ibis in South Florida and captive scarlet ibis (E. ruber) in Brazil. No Plasmodium or Leucocytozoon infections were detected. Parasitemias of the 400 positive birds were very low (average 0.084%, range 0.001%-2.16% [although only 2 birds had parasitemias >1%]). Prevalence and parasitemias were similar for males and females (68% vs. 61.6% and 0.081% vs. 0.071%, respectively). Prevalence in juveniles was lower compared with adults (52% vs. 67.4%) but parasitemias were higher in juveniles (0.117% vs. 0.065%). This data shows that H. plataleae is common in ibis in South Florida. Although parasitemias were generally low, additional research is needed to determine if this parasite has subclinical effects on ibis, if additional haplotypes or parasite species infect ibis in other regions of their range, or if H. plataleae is pathogenic for other sympatric avian species.

Site Fidelity is Associated with Food Provisioning and Salmonella in an Urban Wading Bird.

Food provisioning can change wildlife pathogen dynamics by altering host susceptibility via nutrition and/or through shifts in foraging behavior and space use. We used the American white ibis (Eudocimus albus), a wading bird increasingly observed in urban parks, as a model to study synergistic relationships between food provisioning and infection risk across an urban gradient in South Florida. We tested whether Salmonella prevalence was associated with changes in ibis diet (stable isotope analysis), space use (site fidelity via GPS tracking), and local density (flock size). We compared the relative importance of these mechanisms by ranking candidate models using logistic regression. We detected Salmonella in 27% of white ibises (n = 233) sampled at 15 sites. Ibises with diets higher in anthropogenic food exhibited higher site fidelity. Salmonella prevalence was higher at sites where ibises exhibited greater site fidelity and Salmonella was more prevalent in soil and water. Overlap in Salmonella serotypes between ibises and soil or water also was more likely at sites where ibises exhibited higher site fidelity. Our results suggest that repeated use of foraging areas may increase Salmonella exposure for birds if foraging areas are contaminated from animal feces, human waste, or other bacterial sources. Limiting wildlife feeding in parks-perhaps best achieved through understanding the motivations for feeding, education, and enforcement-may reduce health risks for wildlife and the public.

Surveillance and Genetic Characterization of Virulent Newcastle Disease Virus Subgenotype V.3 in Indigenous Chickens from Backyard Poultry Farms and Live Bird Markets in Kenya.

Kenyan poultry consists of ~80% free-range indigenous chickens kept in small flocks (~30 birds) on backyard poultry farms (BPFs) and they are traded via live bird markets (LBMs). Newcastle disease virus (NDV) was detected in samples collected from chickens, wild farm birds, and other domestic poultry species during a 2017-2018 survey conducted at 66 BPFs and 21 LBMs in nine Kenyan counties. NDV nucleic acids were detected by rRT-PCR L-test in 39.5% (641/1621) of 1621 analyzed samples, of which 9.67% (62/641) were NDV-positive by both the L-test and a fusion-test designed to identify the virulent virus, with a majority being at LBMs (64.5%; 40/62) compared to BPFs (25.5%; 22/62). Virus isolation and next-generation sequencing (NGS) on a subset of samples resulted in 32 complete NDV genome sequences with 95.8-100% nucleotide identities amongst themselves and 95.7-98.2% identity with other east African isolates from 2010-2016. These isolates were classified as a new sub-genotype, V.3, and shared 86.5-88.9% and 88.5-91.8% nucleotide identities with subgenotypes V.1 and V.2 viruses, respectively. The putative fusion protein cleavage site (113R-Q-K-R↓F 117) in all 32 isolates, and a 1.86 ICPI score of an isolate from a BPF chicken that had clinical signs consistent with Newcastle disease, confirmed the high virulence of the NDVs. Compared to genotypes V and VI viruses, the attachment (HN) protein of 18 of the 32 vNDVs had amino acid substitutions in the antigenic sites. A time-scaled phylogeographic analysis suggests a west-to-east dispersal of the NDVs via the live chicken trade, but the virus origins remain unconfirmed due to scarcity of continuous and systematic surveillance data. This study reveals the widespread prevalence of vNDVs in Kenyan backyard poultry, the central role of LBMs in the dispersal and possibly generation of new virus variants, and the need for robust molecular epidemiological surveillance in poultry and non-poultry avian species.

Gut microbiome shifts with urbanization and potentially facilitates a zoonotic pathogen in a wading bird.

Microbial communities in the gastrointestinal tract influence many aspects of host health, including metabolism and susceptibility to pathogen colonization. These relationships and the environmental and individual factors that drive them are relatively unexplored for free-living wildlife. We quantified the relationships between urban habitat use, diet, and age with microbiome composition and diversity for 82 American white ibises (Eudocimus albus) captured along an urban gradient in south Florida and tested whether gut microbial diversity was associated with Salmonella enterica prevalence. Shifts in community composition were significantly associated with urban land cover and, to a lesser extent, diets higher in provisioned food. The diversity of genera was negatively associated with community composition associated with urban land cover, positively associated with age class, and negatively associated with Salmonella shedding. Our results suggest that shifts in both habitat use and diet for urban birds significantly alter gut microbial composition and diversity in ways that may influence health and pathogen susceptibility as species adapt to urban habitats.

The movements of a recently urbanized wading bird reveal changes in season timing and length related to resource use.

The American White Ibis (Eudocimus albus) is a nomadic wading bird that is increasing the amount of time spent foraging in urban areas, relying on artificial wetlands and other anthropogenic resources year-round. In this study, we explore whether and how American White Ibis association with urban environments is predictive of variation in the timing and length of behavioral seasons. Other urbanized species exhibit altered annual cycles such as loss of migratory behavior and year-round breeding related to consistent resource abundance, often related to intentional and unintentional provisioning. To determine if these same patterns of behavior were also present in White Ibis, we used behavioral change point analysis to segment the tracks of 41 ibis equipped with GPS backpacks to identify the initiation and duration of four behavioral seasons (non-breeding, pre-breeding, breeding, post-breeding) the degree of urban association. We found that intraspecific variation in urban habitat use had strong carryover effects on the timing and duration of behavioral seasons. This study revealed ibis with higher use of urban habitats in non-breeding seasons had longer non-breeding seasons and shorter breeding seasons that began earlier in the year compared to ibis that primarily use wetland habitats. The timing and duration of seasons also varied with ibis age, such that ibis spent more time engaged in breeding-related seasons as they aged. Juvenile and subadult ibis, though considered to be reproductively immature, also exhibit behavioral shifts in relation to breeding seasons. The behavioral patterns found in this study provide evidence that ibis are adapting their annual cycles and seasonal behaviors to exploit urban resources. Future research is needed to identify the effect of interactions between ibis urban association and age on behavioral season expression.

EXPERIMENTAL INFECTIONS AND SEROLOGY INDICATE THAT AMERICAN WHITE IBIS (EUDOCIUMUS ALBUS) ARE COMPETENT RESERVOIRS FOR TYPE A INFLUENZA VIRUS.

The American White Ibis (Eudocimus albus) is a nomadic wading bird common to wetland habitats in the southeastern US. In south Florida, US, habitat depletion has driven many ibis to become highly urbanized. Although they forage in neighborhood parks, artificial wetlands, backyards, and golf courses, the majority continue to nest in natural wetlands, often in dense, mixed species colonies. Adults and juveniles commonly disperse thousands of kilometers to other breeding colonies along the Gulf and southeast Atlantic coasts, presenting the potential for close contact with humans, domestic animals, and other wild bird species. Historically, wading birds were not considered to be significant hosts for influenza A virus (IAV), yet as ibis regularly move among various human, domestic animal, and wildlife interfaces, their potential to be exposed to or infected with IAV deserves attention. We experimentally challenged wild-caught, captive-reared White Ibis (n=20) with IAV, tested wild White Ibis for IAV, and serologically tested wild White Ibis for antibodies to IAV. White Ibis were highly susceptible to experimental challenge with H6N1 and H11N9 IAVs, with cloacal shedding lasting an average of 6 d. All 13 infected birds seroconverted by 14 d postinfection as determined by microneutralization. In contrast, no birds challenged with H3N8 were infected. We tested 118 swabs and 578 serum samples from White Ibis captured in southeastern Florida for IAV infection and antibodies to IAV, respectively. Although no IAVs were isolated, 70.4% serum samples were antibody positive by blocking enzyme-linked immunosorbent assay (bELISA). Neutralizing antibodies to H1-H12 were detected in 96.0% of a subset of bELISA positive birds (n=196) and 81.0% tested antibody positive to two or more hemagglutinin subtypes, indicating that exposure to multiple IAVs is common. These results provide evidence that White Ibis are susceptible and naturally infected with IAV and may represent a component of the IAV natural reservoir system.

A retrospective study of Newcastle disease in Kenya.

Newcastle disease (ND) is a major constraint to Kenya's poultry production, which is comprised of approximately 80% indigenous chickens (ICs; caged and free-range system) and 20% exotic chickens (intensive system). This study analyzed cases reported as suspected ND in Kenya between 2005 and 2015. Of the suspected 332 ND reported cases from the three production systems in 27 locations within six Kenyan Agro-Ecological Zones (AEZs), 140 diagnosed as infected with avian orthoavulavirus 1 (AOaV-1; formerly Newcastle disease virus) were present in every year in all AEZs. The numbers of AOaV-1-positive cases differed significantly (p < 0.05) between the production systems across the years depending on the season, climate, and location. In the free-range system, both ambient temperatures and season associated significantly (p = 0.001 and 0.02, respectively) with the number of cases, while in the intensive and caged systems, the positive cases correlated significantly with season and relative humidity, respectively (p = 0.05). Regardless of the production systems, the numbers of clinically sick birds positively correlated with the ambient temperatures (r = 0.6; p < 0.05). Failure to detect AOaV-1 in 58% of the ND cases reported, and mortalities exceeding the observed numbers of clinically sick birds suggest deficiencies in the current ND reporting and diagnostic system. Intensive farmers were the slowest in reporting the cases and diagnostic deficiencies were most evident by failure to test the exposure of ICs to natural infection with AOaV-1 and for the AOaV-1-negative cases lack of testing for other pathogens and/or AOaV-1 variants. This study indicates a need for improved surveillance and diagnostics in Kenyan domestic poultry.

Genetic characterization and pathogenesis of the first H9N2 low pathogenic avian influenza viruses isolated from chickens in Kenyan live bird markets.

Poultry production plays an important role in the economy and livelihoods of rural households in Kenya. As part of a surveillance program, avian influenza virus (AIV)-specific real-time RT-PCR (RRT-PCR) was used to screen 282 oropharyngeal swabs collected from chickens at six live bird markets (LBMs) and 33 backyard poultry farms in Kenya and 8 positive samples were detected. Virus was isolated in eggs from five samples, sequenced, and identified as H9N2 low pathogenic AIV (LPAIV) G1 lineage, with highest nucleotide sequence identity (98.6-99.9%) to a 2017 Ugandan H9N2 isolate. The H9N2 contained molecular markers for mammalian receptor specificity, implying their zoonotic potential. Virus pathogenesis and transmissibility was assessed by inoculating low and medium virus doses of a representative Kenyan H9N2 LPAIV isolate into experimental chickens and exposing them to naïve uninfected chickens at 2 -days post inoculation (dpi). Virus shedding was determined at 2/4/7 dpi and 2/5 days post placement (dpp), and seroconversion determined at 14 dpi/12 dpp. None of the directly-inoculated or contact birds exhibited any mortality or clinical disease signs. All directly-inoculated birds in the low dose group shed virus during the experiment, while only one contact bird shed virus at 2 dpp. Only two directly-inoculated birds that shed high virus titers seroconverted in that group. All birds in the medium dose group shed virus at 4/7 dpi and at 5 dpp, and they all seroconverted at 12/14 dpp. This is the first reported detection of H9N2 LPAIV from Kenya and it was shown to be infectious and transmissible in chickens by direct contact and represents a new disease threat to poultry and potentially to people.

Genomic comparison of Newcastle disease viruses isolated in Nigeria between 2002 and 2015 reveals circulation of highly diverse genotypes and spillover into wild birds.

Newcastle disease virus (NDV) has a wide avian host range and a high degree of genetic variability, and virulent strains cause Newcastle disease (ND), a worldwide concern for poultry health. Although NDV has been studied in Nigeria, genetic information about the viruses involved in the endemicity of the disease and the transmission that likely occurs at the poultry-wildlife interface is still largely incomplete. Next-generation and Sanger sequencing was performed to provide complete (n = 73) and partial genomic sequence data (n = 38) for NDV isolates collected from domestic and wild birds in Nigeria during 2002-2015, including the first complete genome sequences of genotype IV and subgenotype VIh from the African continent. Phylogenetic analysis revealed that viruses of seven different genotypes circulated in that period, demonstrating high genetic diversity of NDV for a single country. In addition, a high degree of similarity between NDV isolates from domestic and wild birds was observed, suggesting that spillovers had occurred, including to three species that had not previously been shown to be susceptible to NDV infection. Furthermore, the first spillover of a mesogenic Komarov vaccine virus is documented, suggesting a previous spillover and evolution of this virus. The similarities between viruses from poultry and multiple bird species and the lack of evidence for host adaptation in codon usage suggest that transmission of NDV between poultry and non-poultry birds occurred recently. This is especially significant when considering that some viruses were isolated from species of conservation concern. The high diversity of NDV observed in both domestic and wild birds in Nigeria emphasizes the need for active surveillance and epidemiology of NDV in all bird species.

Experimental Infection and Transmission of Newcastle Disease Vaccine Virus in Four Wild Passerines.

Our prior work has shown that live poultry vaccines have been intermittently isolated from wild birds sampled during field surveillance studies for Newcastle disease virus (NDV). Thus, we experimentally investigated the susceptibility of four native agriculturally associated wild bird species to the NDV LaSota vaccine and evaluated the shedding dynamics, potential transmission from chickens, and humoral antibody responses. To test susceptibility, we inoculated wild-caught, immunologically NDV-naïve house finches (Haemorhous mexicanus; n = 16), brown-headed cowbirds (Molothrus ater; n = 9), northern cardinals (Cardinalis cardinalis; n = 6), and American goldfinches (Spinus tristis; n = 12) with 0.1 ml (106.7 mean embryo infectious doses [EID50/ml]) of NDV LaSota vaccine via the oculo-nasal route. To test transmission between chickens and wild birds, adult specific-pathogen-free white leghorn chickens were inoculated similarly and cohoused in separate isolators with two to five wild birds of the species listed above. This design resulted in three treatments: wild bird direct inoculation (five groups) and wild bird exposure to one (two groups) or two inoculated chickens (six groups), respectively. Blood and oropharyngeal and cloacal swabs were collected before and after infection with the live vaccine. All wild birds that were directly inoculated with the LaSota vaccine shed virus as demonstrated by virus isolation (VI). Cardinals were the most susceptible species based on shedding viruses from 1 to 11 days postinoculation (dpi) with titers up to 104.9 EID50/ml. Although LaSota viruses were shed by all inoculated chickens and were present in the drinking water, most noninoculated wild birds cohoused with these chickens remained uninfected for 14 days as evidenced by VI. However, one American goldfinch tested positive for vaccine transmission by VI at 7 dpi and one house finch tested positive for vaccine transmission by real-time reverse-transcription PCR at 13 dpi. Only one directly inoculated cowbird (out of three) and two cardinals (out of two) developed NDV-specific hemagglutination inhibition antibody titers of 16, 16, and 128, respectively. No clinical signs were detected in the chickens or the wild birds postinoculation.

Infección experimental y transmisión del virus de la vacuna contra la enfermedad de Newcastle en cuatro paseriformes silvestres. Nuestras investigaciones anteriores han demostrado que las vacunas vivas utilizadas en avicultura se han aislado de forma intermitente de aves silvestres muestreadas durante los estudios de vigilancia en el campo para el virus de la enfermedad de Newcastle (NDV). Por lo tanto, se investigó experimentalmente la susceptibilidad a la vacuna contra la enfermedad de Newcastle cepa LaSota en cuatro especies de aves silvestres y nativas asociadas con se han asociado con la agricultura y se evaluó la dinámica de transmisión, la transmisión potencial desde el pollo y las respuestas de anticuerpos humorales. Para evaluar la susceptibilidad, se inocularon pinzones mexicanos (Haemorhous mexicanus; n = 16), tordos cabecicafés (Molothrus ater; n = 9), cardenales (Cardinalis cardinalis; n = 6) y jilgueros norteamericanos (Spinus tristis; n = 12), todos de origen silvestre y sin exposición previa al virus de Newcastle. Estas aves se inocularon con 0.1 ml (106.7 dosis medias infecciosas para embrión de pollo [EID50]/ml) de la vacuna de Newcastle cepa LaSota a través de la vía oculonasal. Para determinar la transmisión entre pollos y aves silvestres, se inocularon de igual forma aves adulta tipo Leghorn libres de patógenos específicos y se alojaron en unidades de aislamiento en cohabitación con dos a cinco aves silvestres de las especies mencionadas anteriormente. Este diseño dio como resultado tres tratamientos: inoculación directa de aves silvestres (cinco grupos), exposición de aves silvestres a un pollo inoculado (dos grupos), o exposición a dos pollos inoculados (seis grupos), respectivamente. Se recolectaron muestras de sangre e hisopos de la orofaringe y de la cloaca antes y después de la infección con la vacuna viva. Todas las aves silvestres que se inocularon directamente con la vacuna LaSota eliminaron el virus, como se demostró mediante el aislamiento viral (VI). Los cardenales fueron la especie más susceptible con base en el aislamiento viral de uno a 11 días después de la inoculación con títulos de hasta 104.9 EID50/ml. Aunque todos los pollos inoculados eliminaron el virus LaSota y este virus estaba presente en el agua de bebida, la mayoría de las aves silvestres no inoculadas que cohabitaron con estos pollos permanecieron sin infectar durante 14 días, como lo demuestra el aislamiento viral. Sin embargo, un jilguero norteamericano resultó positivo mediante aislamiento viral a la transmisión de la vacuna a los siete días después de la inoculación y un pinzón mexicano resultó positivo para la transmisión de la vacuna mediante transcripción reversa y PCR en tiempo real a los 13 días después de la inoculación. Solo un tordo cabecicafé inoculado directamente (de un total de tres) y dos cardenales (de un total de dos) desarrollaron títulos de anticuerpos de inhibidores de la hemaglutinación específicos contra la enfermedad de Newcastle de 16, 16 y 128, respectivamente. No se detectaron signos clínicos en los pollos ni en las aves silvestres después de la inoculación.

From wetland specialist to hand-fed generalist: shifts in diet and condition with provisioning for a recently urbanized wading bird.

Many wildlife species shift their diets to use novel resources in urban areas. The consequences of these shifts are not well known, and consumption of reliable-but low quality-anthropogenic food may present important trade-offs for wildlife health. This may be especially true for carnivorous species such as the American white ibis (Eudocimus albus), a nomadic wading bird which has been increasingly observed in urban parks in South Florida, USA. We tested the effects of anthropogenic provisioning on consumer nutrition (i.e. dietary protein), body condition and ectoparasite burdens along an urban gradient using stable isotope analysis, scaled mass index values and GPS transmitter data. Ibises that assimilated more provisioned food were captured at more urban sites, used more urban habitat, had lower mass-length residuals, lower ectoparasite scores, assimilated less δ15N and had smaller dietary isotopic ellipses. Our results suggest that ibises in urban areas are heavily provisioned with anthropogenic food, which appears to offer a trade-off by providing low-quality, but easily accessible, calories that may not support high mass but may increase time available for anti-parasite behaviours such as preening. Understanding such trade-offs is important for investigating the effects of provisioning on infection risk and the conservation of wildlife in human-modified habitats.This article is part of the theme issue 'Anthropogenic resource subsidies and host-parasite dynamics in wildlife'.

Assessing the contributions of intraspecific and environmental sources of infection in urban wildlife: Salmonella enterica and white ibis as a case study.

Conversion of natural habitats into urban landscapes can expose wildlife to novel pathogens and alter pathogen transmission pathways. Because transmission is difficult to quantify for many wildlife pathogens, mathematical models paired with field observations can help select among competing transmission pathways that might operate in urban landscapes. Here we develop a mathematical model for the enteric bacteria Salmonella enterica in urban-foraging white ibis ( Eudocimus albus) in south Florida as a case study to determine (i) the relative importance of contact-based versus environmental transmission among ibis and (ii) whether transmission can be supported by ibis alone or requires external sources of infection. We use biannual field prevalence data to restrict model outputs generated from a Latin hypercube sample of parameter space and select among competing transmission scenarios. We find the most support for transmission from environmental uptake rather than between-host contact and that ibis-ibis transmission alone could maintain low infection prevalence. Our analysis provides the first parameter estimates for Salmonella shedding and uptake in a wild bird and provides a key starting point for predicting how ibis response to urbanization alters their exposure to a multi-host zoonotic enteric pathogen. More broadly, our study provides an analytical roadmap to assess transmission pathways of multi-host wildlife pathogens in the face of scarce infection data.

Diversity and prevalence of hemoparasites of wading birds in southern Florida, USA.

Relatively few studies on hemoparasites have been conducted on wading birds in the families Ardeidae and Threskiornithidae (order Pelecaniformes), especially in the United States. In this study, we obtained baseline data on the prevalence and genetic diversity of haemosporidian parasites in wading birds opportunistically sampled from southern Florida, USA. We detected blood parasites in White Ibis (Eudocimus albus), Glossy Ibis (Plegadis falcinellus), Green Heron (Butorides virescens), and Roseate Spoonbill (Platalea ajaja) with several novel host-parasite relationships. Infected birds had low parasitemias (average 0.77%, range 0-4%) suggesting that infections were chronic. Despite the low sample sizes for several of our sampled species, these data highlight the diversity of parasites in this understudied group of birds and suggest that additional studies are needed to investigate the potential impacts of these parasites on their health, especially since southern Florida is becoming increasingly urbanized which can alter parasite transmission or host susceptibility.

Urbanized White Ibises (Eudocimus albus) as Carriers of Salmonella enterica of Significance to Public Health and Wildlife.

Worldwide, Salmonella spp. is a significant cause of disease for both humans and wildlife, with wild birds adapted to urban environments having different opportunities for pathogen exposure, infection, and transmission compared to their natural conspecifics. Food provisioning by people may influence these factors, especially when high-density mixed species flocks aggregate. White Ibises (Eudocimus albus), an iconic Everglades species in decline in Florida, are becoming increasingly common in urbanized areas of south Florida where most are hand-fed. We examined the prevalence of Salmonella shedding by ibises to determine the role of landscape characteristics where ibis forage and their behavior, on shedding rates. We also compared Salmonella isolated from ibises to human isolates to better understand non-foodborne human salmonellosis. From 2010-2013, 13% (n = 261) adult/subadult ibises and 35% (n = 72) nestlings sampled were shedding Salmonella. The prevalence of Salmonella shedding by ibises significantly decreased as the percent of Palustrine emergent wetlands and herbaceous grasslands increased, and increased as the proportion of open-developed land types (e.g. parks, lawns, golf courses) increased, suggesting that natural ecosystem land cover types supported birds with a lower prevalence of infection. A high diversity of Salmonella serotypes (n = 24) and strain types (43 PFGE types) were shed by ibises, of which 33% of the serotypes ranked in the top 20 of high significance for people in the years of the study. Importantly, 44% of the Salmonella Pulsed-Field Gel Electrophoresis patterns for ibis isolates (n = 43) matched profiles in the CDC PulseNet USA database. Of these, 20% came from Florida in the same three years we sampled ibis. Importantly, there was a negative relationship between the amount of Palustrine emergent wetland and the number of Salmonella isolates from ibises that matched human cases in the PulseNet database (p = 0.056). Together, our results indicate that ibises are good indicators of salmonellae strains circulating in their environment and they have both the potential and opportunity to transmit salmonellae to people. Finally, they may act as salmonellae carriers to natural environments where other more highly-susceptible groups (nestlings) may be detrimentally affected.