Epidemiological investigation of fowl adenovirus (FAdV) infections in ducks and geese in Shandong Province, China.

RESEARCH HIGHLIGHTS: Samples of suspected FAdV-infected waterfowl from farms in Shandong Province were collected from 2019 to 2022.Single infections with FAdV were less frequent than mixed infections.477 out of 792 samples (60.23%) tested positive for FAdV nucleic acids.Detection rate of FAdV was 65.47% in fattening duck farms, 55.73% in breeder duck farms and 54.55% in fattening geese farms.

The habitat preferences of invasive raccoon dog imply elevated risks for wetland-associated prey species.

Habitat preferences of invasive predators determine where and for which prey species they pose a threat upon. This is crucial information for the conservation of endangered prey species because invasive predators pose additional predation on top of that caused by natural predators. In large parts of Europe, the most common invasive mesopredator is the raccoon dog (Nyctereutes procyonoides). To understand the risk that the raccoon dog poses for wetland-associated species, we collected information about its habitat preferences near these habitats. We used data on 24 GPS-collared raccoon dogs from three landscape types in Finland, to study their spatial and temporal habitat preferences. We first determined their home ranges, within which we then examined habitat use and preferences. Raccoon dogs showed generalist habitat use, which was evident in their wide range of used habitats. However, in spring and summer, during the breeding seasons of waterfowl and amphibians, they preferred wetlands and peatlands. They also preferred shorelines and the edges of forests and agricultural fields. During autumn and winter, raccoon dogs did not prefer wetlands. These findings support the conclusion that the raccoon dog's habitat preferences pose a particular threat to wetland-associated species, such as nesting waterfowl and amphibians. The species' habitat preferences coupled with high numbers of this invasive mesopredator pose additional predation for endangered wetland-associated species on top of that of native predators.

Systematics and life cycles of four avian schistosomatids from Southern Cone of South America.

Relative to the numerous studies focused on mammalian schistosomes, fewer include avian schistosomatids particularly in the southern hemisphere. This is changing and current research emerging from the Neotropics shows a remarkable diversity of endemic taxa. To contribute to this effort, nine ducks (Spatula cyanoptera, S.versicolor, Netta peposaca), 12 swans (Cygnus melancoryphus) and 1,400 Physa spp. snails from Chile and Argentina were collected for adults and larval schistosomatids, respectively. Isolated schistosomatids were preserved for morphological and molecular analyses (28S and COI genes). Four different schistosomatid taxa were retrieved from birds: Trichobilharzia sp. in N. peposaca and S. cyanoptera that formed a clade; S.cyanoptera and S. versicolor hosted Trichobilharzia querquedulae; Cygnus melancoryphus hosted the nasal schistosomatid, Nasusbilharzia melancorhypha; and one visceral, Schistosomatidae gen. sp., which formed a clade with furcocercariae from Argentina and Chile from previous work. Of the physid snails, only one from Argentina had schistosomatid furcocercariae that based on molecular analyses grouped with T. querquedulae. This study represents the first description of adult schistosomatids from Chile as well as the elucidation of the life cycles of N.melancorhypha and T. querquedulae in Chile and Neotropics, respectively. Without well-preserved adults, the putative new genus Schistosomatidae gen. sp. could not be described, but its life cycle involves Chilina spp. and C. melancoryphus. Scanning electron microscopy of T. querquedulae revealed additional, undescribed morphological traits, highlighting its diagnostic importance. Authors stress the need for additional surveys of avian schistosomatids from the Neotropics to better understand their evolutionary history.

Never be mute about bird welfare: Swanning around with environmental enrichment.

Environmental enrichment (EE) is commonly provided to animals managed under human care, being beneficial to behavioral diversity and improving animal welfare. Use of EE appears to be particularly beneficial to individual wild animals spending a short period of time in captivity, for example, as part of conservation or rehabilitation programs. This paper documents a case study on the application and relevance of EE for a group of captive mute swans housed in a rescue center. Observational data were analyzed for two groups of juvenile swans that were provided with a physical EE device to increase time spent foraging. Periods of no EE were observed and compared to data from when birds were provided with EE. Results show that EE promoted foraging time and helped to reduce long periods of inactivity in captive birds. EE helped to reduce occurrence of captive-focused (i.e., abnormal behaviors) although these was already seen at very low rates. Inactivity as a measure of welfare in captive swans specifically (and waterbirds generally) should be further investigated to understand potential impacts on bird health. Our research shows the benefits of simple and easy-to-use EE devices on captive animal behavior and how use of EE for individuals spending a short amount of time in captivity (e.g., within a rescue center) could ensure diversity of behavior patterns and promote the performance of adaptive behaviors upon release to the wild.

A gradual migratory divide determines not only the direction of migration but also migration strategy of a social migrant bird.

Migratory divides separate populations of migratory animals, facilitating the evolution of intraspecific differences in migration strategies. Migration strategies are expected to be different for birds using different flyways and environments, but the knowledge regarding the impact of the flyway on individual migration strategies is scarce. By using satellite tracking and neckband resightings, we reveal the existence and structure of a gradual migratory divide between two European flyway populations of greylag geese Anser anser. Birds breeding at the far end of the Gulf of Bothnia in the Baltic Sea coast use the Western Flyway, those breeding in the Gulf of Finland the Central Flyway and those breeding between these extremes scatter to the two flyways. By using Gaussian process modelling, we show that migration strategies differed between the flyways. The birds using the Western Flyway migrated earlier in autumn, performed longer annual migration and made a clear stopover during migration, whereas the birds using the Central Flyway flew directly to their wintering sites. The gradual migratory divide that also divides migration strategies provides insights into migratory divides on birds with learned migration. Distinct migration strategies in different flyways provide exciting possibilities to further study the factors driving migration strategies.

Population genomics indicate three different modes of divergence and speciation with gene flow in the green-winged teal duck complex.

The processes leading to divergence and speciation can differ broadly among taxa with different life histories. We examine these processes in a small clade of ducks with historically uncertain relationships and species limits. The green-winged teal (Anas crecca) complex is a Holarctic species of dabbling duck currently categorized as three subspecies (Anas crecca crecca, A. c. nimia, and A. c. carolinensis) with a close relative, the yellow-billed teal (Anas flavirostris) from South America. A. c. crecca and A. c. carolinensis are seasonal migrants, while the other taxa are sedentary. We examined divergence and speciation patterns in this group, determining their phylogenetic relationships and the presence and levels of gene flow among lineages using both mitochondrial and genome-wide nuclear DNA obtained from 1,393 ultraconserved element (UCE) loci. Phylogenetic relationships using nuclear DNA among these taxa showed A. c. crecca, A. c. nimia, and A. c. carolinensis clustering together to form one polytomous clade, with A. flavirostris sister to this clade. This relationship can be summarized as (crecca, nimia, carolinensis)(flavirostris). However, whole mitogenomes revealed a different phylogeny: (crecca, nimia)(carolinensis, flavirostris). The best demographic model for key pairwise comparisons supported divergence with gene flow as the probable speciation mechanism in all three contrasts (crecca-nimia, crecca-carolinensis, and carolinensis-flavirostris). Given prior work, gene flow was expected among the Holarctic taxa, but gene flow between North American carolinensis and South American flavirostris (M âˆ¼0.1-0.4 individuals/generation), albeit low, was not expected. Three geographically oriented modes of divergence are likely involved in the diversification of this complex: heteropatric (crecca-nimia), parapatric (crecca-carolinensis), and (mostly) allopatric (carolinensis-flavirostris). Our study shows that ultraconserved elements are a powerful tool for simultaneously studying systematics and population genomics in systems with historically uncertain relationships and species limits.

Waterfowl show spatiotemporal trends in influenza A H5 and H7 infections but limited taxonomic variation.

Influenza A viruses in wild birds pose threats to the poultry industry, wild birds, and human health under certain conditions. Of particular importance are wild waterfowl, which are the primary reservoir of low-pathogenicity influenza viruses that ultimately cause high-pathogenicity outbreaks in poultry farms. Despite much work on the drivers of influenza A virus prevalence, the underlying viral subtype dynamics are still mostly unexplored. Nevertheless, understanding these dynamics, particularly for the agriculturally significant H5 and H7 subtypes, is important for mitigating the risk of outbreaks in domestic poultry farms. Here, using an expansive surveillance database, we take a large-scale look at the spatial, temporal, and taxonomic drivers in the prevalence of these two subtypes among influenza A-positive wild waterfowl. We document spatiotemporal trends that are consistent with past work, particularly an uptick in H5 viruses in late autumn and H7 viruses in spring. Interestingly, despite large species differences in temporal trends in overall influenza A virus prevalence, we document only modest differences in the relative abundance of these two subtypes and little, if any, temporal differences among species. As such, it appears that differences in species' phenology, physiology, and behaviors that influence overall susceptibility to influenza A viruses play a much lesser role in relative susceptibility to different subtypes. Instead, species are likely to freely pass viruses among each other regardless of subtype. Importantly, despite the similarities among species documented here, individual species still may play important roles in moving viruses across large geographic areas or sustaining local outbreaks through their different migratory behaviors.

Migratory connectivity of North American waterfowl across administrative flyways.

Management of waterfowl that migrate seasonally across North America occurs within four flyways that were delineated in the early 1900s to include the annual movements of populations. Movements may have changed over the past century since the administrative flyways were established, and may do so while management plans are in use, so information about transitions among flyways through time can illustrate how management assumptions may change. Today there are more than 12 million records from 60 years of migratory waterfowl band recoveries to assess adaptive management approaches that will be most effective when they account for movements within and between flyways. We examined how much the movement of North American waterfowl occurs between flyways, whether those movements have changed through time, and whether movements of mallards are representative of multiple species, as suggested by current harvest management strategies. We estimated the probability a duck would transition from one flyway to another and the strength of migratory connectivity (MC) for each species within and among flyways. We used capture-mark-recovery models to estimate population-specific movement within and among flyways (transition probabilities) for 15 migratory waterfowl species that were banded during breeding and recovered during winter. We developed new functionality in the R package MigConnectivity to estimate the species-specific strength of MC using transition probability samples from the capture-mark-recovery models. We found the regular movement of duck populations among flyways, overall weak MC, and no consistent change in migratory movements through time. Mallard movements were median among all duck species, but significantly different from many species, particularly diving ducks. Despite the significant movement between flyways, our work suggests flyway management of waterfowl matches many of the seasonal movements of these species when considering mid-continent flyway management. We recommend models accounting for all transition probabilities between populations and regularly estimating harvest derivations, transition probabilities, and MC metrics to verify that the current movements match model assumptions.

A life-history spectrum of population responses to simultaneous change in climate and land use.

Climate and land use change are two of the primary threats to global biodiversity; however, each species within a community may respond differently to these facets of global change. Although it is typically assumed that species use the habitat that is advantageous for survival and reproduction, anthropogenic changes to the environment can create ecological traps, making it critical to assess both habitat selection (e.g. where species congregate on the landscape) and the influence of selected habitats on the demographic processes that govern population dynamics. We used a long-term (1958-2011), large-scale, multi-species dataset for waterfowl that spans the United States and Canada to estimate species-specific responses to climate and land use variables in a landscape that has undergone significant environmental change across space and time. We first estimated the effects of change in climate and land use variables on habitat selection and population dynamics for nine species. We then hypothesized that species-specific responses to environmental change would scale with life-history traits, specifically: longevity, nesting phenology and female breeding site fidelity. We observed species-level heterogeneity in the demographic and habitat selection responses to climate and land use change, which would complicate community-level habitat management. Our work highlights the importance of multi-species monitoring and community-level analysis, even among closely related species. We detected several relationships between life-history traits, particularly nesting phenology, and species' responses to environmental change. One species, the early-nesting northern pintail (Anas acuta), was consistently at the extreme end of responses to land use and climate predictors and has been a species of conservation concern since their population began to decline in the 1980s. They, and the blue-winged teal, also demonstrated a positive habitat selection response to the proportion of cropland on the landscape that simultaneously reduced abundance the following year, indicative of susceptibility to ecological traps. By distilling the diversity of species' responses to environmental change within a community, our methodological approach and findings will help improve predictions of community responses to global change and can inform multi-species management and conservation plans in dynamic landscapes that are based on simple tenets of life-history theory.

Quantifying population-level conservation impacts for a perpetual conservation program on private land.

Area-based targets, such as percentages of regions protected, are popular metrics of success in the protection of nature. While easily quantified, these targets can be uninformative about the effectiveness of conservation interventions and should be complemented by program impact evaluations. However, most impact evaluations have examined the effect of protected areas on deforestation. Studies that have extended these evaluations to more dynamic systems or different outcomes are less common, largely due to data availability. In these cases, simulations might prove to be a valuable tool for gaining an understanding of the potential range of program effect sizes. Here, we employ simulations of wetland drainage to estimate the impact of the United States Fish and Wildlife Service Small Wetlands Acquisition Program (SWAP) across a ten-year period in terms of wetland area, and breeding waterfowl and brood abundance in the Prairie Pothole Region of North Dakota, South Dakota, and Montana. Using our simulation results, we estimate a plausible range of program impact for the SWAP as an avoided loss of between 0.00% and 0.02% of the carrying capacity for broods and breeding waterfowl from 2008-2017. Despite the low programmatic impact that these results suggest, the perpetual nature of SWAP governance provides promising potential for a higher cumulative conservation impact in the long term if future wetland drainage occurs.

Effects of particle size of corn and stocking densities on the performance, carcass traits and gastrointestinal tract development of Muscovy ducks in housing.

The current investigation was conducted to investigate the effects of different particle sizes of corn and stocking densities on performance, carcass traits and gastrointestinal tract (GIT) development of Muscovy ducks in housing. 200 Muscovy ducks were distributed in a factorial scheme (2 × 2) where the treatments were constituted by two stocking densities (2 or 3 birds/m2) and two particle sizes of corn (6 or 8 mm), with five replicates of 10 birds each. At 1, 35, 70, and 90 days old, Muscovy ducks were slaughtered to evaluate the development of heart and GIT. Muscovy ducks managed in the density of 3 birds/m2 presented higher (P < 0.05) GIT development, performance and carcass traits regardless of age evaluated. In the same way, Muscovy ducks fed diets using a particle size of corn of 6 mm (Mean Geometric Diameter (MGD) = 781 µm) presented higher (P < 0.05) GIT development, performance and carcass traits regardless of age evaluated. Conclusively, the stocking density of 3 birds/m2 and the particle size of corn of 6 mm (MGD = 781 µm) were considered ideal recommendations for Muscovy ducks, providing higher GIT development, which resulted in better performance and carcass traits.

Successful Surgical Removal of the Crystalline Lenses in a Black-Necked Swan (Cygnus melancoryphus).

A captive-bred, adult, male, black-necked swan (Cygnus melancoryphus) was presented for evaluation of apparent vision loss due to cataract formation of an unknown duration. The animal was having difficulty navigating its enclosure, and lenticular opacities had been previously noted in both eyes. On examination, bilateral hypermature cataracts were diagnosed. Following preoperative diagnostic testing, surgical removal of the crystalline lenses in both eyes was performed using minor modifications of standard techniques. Follow-up examination and behavioral observation at 60 days postsurgery indicated that vision had been successfully restored without complications. We conclude that successful surgical removal of cataracts is possible in this species using modifications of standard techniques.

Waterfowl Wintering in Cities of Temperate Climatic Zone and Its Shaping Factors.

The dynamics of the population size and species diversity of wintering waterfowl was for the first time studied in 30 cities of Russia over eight years. The total population size increased by 35%. The species diversity was found to reach 32 species in total. The mallard Anas platyrhynchos dominated in all 30 cities. The cities grouped to form seven clusters significantly differing from each other by the total parameter set. The geographical location and climatic characteristics of a city are the main environmental conditions for both people and birds. The variance of the wintering waterfowl population size was determined to the extent of 93% by the area and human population size of the city; and 62% of the variance in species number was determined by the longitude, the average January temperature, and the human population size of the city (p < 0.001). The population size and species number of birds correlated with the "age" of the wintering place (p < 0.05). If the urbanization rate continues the same, the population size and species diversity are expected to grow further.

Citizen science reveals waterfowl responses to extreme winter weather.

Global climate change is increasing the frequency and severity of extreme climatic events (ECEs) which may be especially detrimental during late-winter when many species are surviving on scarce resources. However, monitoring animal populations relative to ECEs is logistically challenging. Crowd-sourced datasets may provide opportunity to monitor species' responses to short-term chance phenomena such as ECEs. We used 14 years of eBird-a global citizen science initiative-to examine distribution changes for seven wintering waterfowl species across North America in response to recent extreme winter polar vortex disruptions. To validate inferences from eBird, we compared eBird distribution changes against locational data from 362 GPS-tagged Mallards (Anas platyrhynchos) in the Mississippi Flyway. Distributional shifts between eBird and GPS-tagged Mallards were similar following an ECE in February 2021. In general, the ECE affected continental waterfowl population distributions; however, responses were variable across species and flyways. Waterfowl distributions tended to stay near wintering latitudes or moved north at lesser distances compared with non-ECE years, suggesting preparedness for spring migration was a stronger "pull" than extreme weather was a "push" pressure. Surprisingly, larger-bodied waterfowl with grubbing foraging strategies (i.e., geese) delayed their northward range shift during ECE years, whereas smaller-bodied ducks were less affected. Lastly, wetland obligate species shifted southward during ECE years. Collectively, these results suggest specialized foraging strategies likely related to resource limitations, but not body size, necessitate movement from extreme late-winter weather in waterfowl. Our results demonstrate eBird's potential to monitor population-level effects of weather events, especially severe ECEs. eBird and other crowd-sourced datasets can be valuable to identify species which are adaptable or vulnerable to ECEs and thus, begin to inform conservation policy and management to combat negative effects of global climate change.

First detection of avian metapneumovirus subtype C Eurasian lineage in a Eurasian wigeon (Mareca penelope) wintering in Northeastern Italy: an additional hint on the role of migrating birds in the viral epidemiology.

Avian metapneumovirus (aMPV) economically affects the global poultry industry causing respiratory and reproductive disorders. Considering the paucity of data on aMPV occurrence in European free-ranging avifauna, a molecular survey was conducted on wild birds of 23 species belonging to the orders Anseriformes, Charadriiformes or Passeriformes, captured alive and sampled in Northeast Italy as part of the national avian influenza virus (AIV) surveillance activities. A total of 492 oropharyngeal swabs, collected from 2007-2010, all AIV-negative, were screened from aMPV by subtype-specific qRT-PCR. An aMPV-C strain, named aMPV/C/IT/Wigeon/758/07, was found in a wintering young Eurasian wigeon (Mareca penelope) sampled in November 2007. The matrix, fusion, and attachment glycoprotein genes of the detected strain were subsequently amplified by specific independent RT-PCRs, then sequenced, and compared in a phylogenetic framework with known aMPV homologous sequences retrieved from GenBank. Close genetic relationships were found between the aMPV/C/IT/Wigeon/758/07 strain and subtype C Eurasian lineage strains isolated in the late 1990s in French domestic ducks, suggesting epidemiological links. Eurasian wigeons are medium/long-range migrant dabbling ducks that move along the Black Sea/Mediterranean flyway; our finding might, therefore, be related to migratory bridges between countries. To our knowledge, this is the first molecular evidence of the occurrence of aMPV subtype C in Italy and backdates the aMPV-C circulation to 2007. Moreover, the results suggest the susceptibility of Eurasian wigeons to aMPV. Broader investigations are needed to assess the role of wild ducks and the significance of the wildfowl/poultry interface in aMPV-C epidemiology.RESEARCH HIGHLIGHTSWild birds live-captured in Italy were tested for aMPV detection and characterization.aMPV-C Eurasian lineage was found for the first time in a wintering Eurasian wigeon.Migratory birds could be involved in the aMPV epidemiology.

Development and evaluation of temperature-sensitive Mycoplasma anserisalpingitidis clones as vaccine candidates.

Mycoplasma anserisalpingitidis is economically the most important pathogenic Mycoplasma species of waterfowl in Europe and Asia. The lack of commercially available vaccines against M. anserisalpingitidis had prompted this study with the aim to produce temperature-sensitive (ts+) clones as candidates for an attenuated live vaccine. The production of ts+ clones was performed by N-methyl-N'-nitro-N-nitrosoguanidine (NTG)-induced mutagenesis of Hungarian M. anserisalpingitidis field isolates. The clones were administered via eye-drop and intracloacally to 33-day-old geese. Colonization ability was examined by PCR and isolation from the trachea and cloaca, while the serological response of the birds was tested by ELISA. Pathological and histopathological examinations were performed in the eighth week after inoculation. Whole-genome sequence (WGS) analysis of the selected clone and its parent strain was also performed. NTG-treatment provided three ts+ mutants (MA177/1/11, MA177/1/12, MA271). MA271 was detected at the highest rate from cloacal (86.25%) and tracheal (30%) samples, while MA177/1/12 and MA271 elicited remarkable serological responses with 90% of the birds showing seroconversion. Re-isolates of MA271 remained ts+ throughout the experiment. Based on these properties, clone MA271 was found to be the most promising vaccine candidate. WGS analysis revealed 59 mutations in the genome of MA271 when compared to its parent strain, affecting both polypeptides involved in different cellular processes and proteins previously linked to bacterial fitness and virulence. Although further studies are needed to prove that MA271 is in all aspects a suitable vaccine strain, it is expected that this ts+ clone will contribute to the control of M. anserisalpingitidis infection.RESEARCH HIGHLIGHTS Three M. anserisalpingitidis ts+ vaccine candidates were produced by NTG-mutagenesis.Clone MA271 was able to colonize geese and induce a serological response.MA271 re-isolates remained ts+ during the 8-week-long experiment.WGS analysis revealed 59 mutations in the genome of MA271.

Pharmacokinetics of ivermectin after oral and intravenous administration in Bilgorajska geese (Anser anser domesticus).

AIMS: To assess the pharmacokinetic profile of ivermectin in Bilgorajska geese (Anser anser domesticus) after single I/V or oral administration, in order to compare these routes of administration and assess oral bioavailability. METHODS: Ten healthy male geese were used in a single-dose, two-phase study with a 3-month washout period between phases. In the first phase, all geese were given 0.2 mg/kg I/V ivermectin, while in the second phase they were treated orally with the same dosage. Blood samples were collected at selected time points up to 480 hours after each administration. Samples were purified using protein precipitation and drug concentration was quantified using HPLC. The analytical method was validated on blank goose plasma and was characterised by an optimal linearity and a limit of quantification of 0.025 µg/mL. The pharmacokinetic analysis was carried out using a non-compartmental approach. RESULTS: The drug was quantifiable up to 240 hours after I/V administration, while after oral treatment it was quantifiable up to 144 hours in most of the geese. The elimination half-life of ivermectin was approximately 3.8 (95% CI = 1.98-7.92; p = 0.027) times higher after I/V administration compared to oral administration. Moreover, the area under the curve from zero to the last detectable timepoint was 6.4 (95% CI = 4.65-8.74; p < 0.001) hours greater after I/V than oral administration. This difference led to a bioavailability of 20.38 (SD 5.92) %. CONCLUSIONS: Following oral administration in geese, ivermectin has a bioavailability of approximately 20%. Further research on the action of ivermectin in the gastrointestinal tract is required along with assessment of tissue residues to allow calculation of withdrawal time to ensure consumer safety. ABBREVIATIONS: AUC: Area under the concentration-time curve; AUClast: Area under the curve from zero to the last detectable timepoint; AUMC: Area under the first moment curve; Cmax: Maximum concentration; Tmax: Time at maximum plasma concentration.

Pseudocorynosoma enrietti (Molfi & Freitas Fernandes, 1953) (Acanthocephala: Polymorphidae) from Patagonia (Argentina): life cycle, localities, and new host records.

All Pseudocorynosoma species inhabit freshwater environments of the American continent, but little is known about their life cycles. We report Pseudocorynosoma enrietti (Molfi & Freitas Fernandes, 1953) from natural and experimental specimens in Patagonia and identify the intermediate and definitive hosts of its life cycle for the first time in South America. Adult worms were recovered from Anas platyrhynchos (Linnaeus) and from a new definitive host, Coscoroba coscoroba Molina. Naturally infected amphipods, Hyalella patagonica Ortmann, were collected to obtain cystacanths that were fed to Gallus gallus domesticus (Linnaeus) and Anas platyrhynchos. Specimens of P. enrietti are described in detail using light and scanning electron microscopy. A key to species of the genus Pseudocorynosoma is included. Worms are characterized in both sexes by fore-trunk spines, and genital spines in an isolated field. The proboscis has 19-20 hook rows; males have 9-11 (10) hooks per row and females 7-9 (8). Males with four cement glands similar in size. Eggs elongated, with filaments. Experimental male and female worms were recovered from A. platyrhynchos at seven and 14 days, post-infection.

Biofilm formation and its impact on environmental survival and antibiotic resistance of Mycoplasma anserisalpingitidis strains.

Several Mycoplasma species can form biofilm, facilitating their survival in the environment, and shielding them from therapeutic agents. The aim of this study was to examine the biofilm-forming ability and its potential effects on environmental survival and antibiotic resistance in Mycoplasma anserisalpingitidis, the clinically and economically most important waterfowl Mycoplasma species. The biofilm-forming ability of 32 M. anserisalpingitidis strains was examined by crystal violet assay. Biofilms and planktonic cultures of the selected strains were exposed to a temperature of 50 °C (20 and 30 min), to desiccation at room temperature (16 and 24 h), or to various concentrations of eight different antibiotics. Crystal violet staining revealed great diversity in the biofilm-forming ability of the 32 tested M. anserisalpingitidis strains, with positive staining in more than half of them. Biofilms were found to be more resistant to heat and desiccation than planktonic cultures, while no correlation was shown between biofilm formation and antibiotic susceptibility. Our results indicate that M. anserisalpingitidis biofilms may contribute to the persistence of the organisms in the environment, which should be taken into account for proper management. Antibiotic susceptibility was not affected by biofilm formation; however, it is important to note that correlations were examined only in vitro.

Subtype Diversity of Influenza A Virus in North American Waterfowl: a Multidecade Study.

The discovery in 1976 of waterfowl as the primary reservoir of influenza A viruses (IAVs) has since spurred decades of waterfowl surveillance efforts by researchers dedicated to understanding the ecology of IAV and its subsequent threat to human and animal health. Here, we employed a multidecade, continental-scale approach of surveillance data to understand trends of seasonal IAV subtype diversity. Between 1976 and 2015, IAVs were detected in 8,427 (10.8%) of 77,969 samples from migratory waterfowl throughout the Central and Mississippi Migratory Flyways in the United States and Canada. A total of 96 hemagglutinin (HA)/neuraminidase (NA) subtype combinations were isolated, which included most HA (H1 to H14) and all 9 NA subtypes. We observed an annual trend of high influenza prevalence, involving a few dominant subtypes, on northern breeding grounds during summer with progressively lowered influenza prevalence, comprised of a highly diverse profile of subtypes, as waterfowl migrate toward southern wintering grounds. Isolates recovered during winter had the highest proportion of mixed and rare HA/NA combinations, indicating increased opportunity for reassortment of IAVs. In addition, 70% of H5 and 49% of H7 IAV isolates were recovered from samples collected during fall and spring, respectively; these are subtypes that can have significant implications for public health and agriculture sectors. Annual cyclical dominance of subtypes on northern breeding grounds is revealed through the longitudinal nature of this study. Our novel findings exhibit the unrealized potential for discovery using existing IAV surveillance data.IMPORTANCE Wild aquatic birds are the primary natural reservoir of influenza A viruses (IAVs) and are therefore responsible for the dispersal and maintenance of IAVs representing a broad range of antigenic and genetic diversity. The aims of IAV surveillance in waterfowl not only relate to understanding the risk of spillover risk to humans, but also to improving our understanding of basic questions related to IAV evolution and ecology. By evaluating several decades of surveillance data from wild aquatic birds sampled along North American migratory flyways, we discovered an annual trend of increasing subtype diversity during southbound migration, peaking on southern wintering grounds. Winter sampling revealed the highest proportion of mixed and rare infections that suggest higher opportunity for spillover. These findings allow improvements to surveillance efforts to robustly capture IAV diversity that will be used for vaccine development and cultivate a more thorough understanding of IAV evolution and persistence mechanisms.