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Understanding the formation of multiple populations in globular clusters (GCs) represents a challenge for stellar population studies. Nevertheless, the outermost cluster regions, likely to hold clues about the initial configuration of GC stars, remain underexplored. We use synthetic spectra reflecting the chemical compositions of first- and second-population (1P, 2P) stars in 47 Tucanae to identify spectral regions sensitive to these populations. This led us to define new photometric bands that effectively distinguish 1P and 2P giant stars using Gaia XP spectra. Testing these filters, we constructed the pseudo two-colour diagrams dubbed chromosome maps (ChMs) and, for the first time, identified 1P and 2P stars in the cluster's outermost regions and beyond its tidal radius. We constructed similar diagrams for NGC 3201, NGC 6121, NGC 6752, and NGC 6397, thus exploring GCs with different metallicities. The ChMs effectively distinguished multiple populations in the outer regions of all clusters, except for the metal-poor NGC 6397. Our findings, together with literature results from more-internal regions, show that the 2P stars of 47 Tucanae are more-centrally concentrated than the 1P. A similar pattern is seen for 2P stars with extreme chemical composition of NGC 3201. The multiple populations of NGC 6121, and NGC 6752 share the same radial distributions. These radial behaviours are consistent with the GC formation scenarios where 2P stars originate in the central regions. Noticeably, results on NGC 3201 are in tension with the conclusion from recent work that its 1P is more centrally concentrated than the 2P and might form with more central concentration. [ABSTRACT FROM AUTHOR]

Globular cluster (GC) stars composed of pristine material, also known as first-generation (1G) stars, are not chemically homogeneous as they exhibit extended sequences in the chromosome map (ChM). Recent studies characterized 1G stars within the center of 55 Galactic GCs, revealing metallicity variations. Despite this progress, several unanswered questions persist, particularly concerning the link between the 1G metallicity spread and factors such as the radial distance from the cluster center or the host GC parameters. Additionally, it remains unclear whether the extended 1G sequence phenomenon is exclusive to old Galactic GCs with multiple populations. This work addresses these open issues, examining 1G stars in different environments. First, we combine Hubble Space Telescope (HST) and James Webb Space Telescope photometry of the GC 47 Tucanae to study 1G stars at increasing distances from the cluster center. We find that metal-rich 1G stars are more centrally concentrated than metal-poor ones, suggesting a metallicity radial gradient. Additionally, the two groups of 1G stars share similar kinematics. Since our analysis focuses on giant stars in the cluster center and M dwarfs in external fields, we discuss the possibility that the metallicity distribution depends on stellar mass. Subsequently, we analyze HST multi-band photometry of two simple-population clusters, NGC 6791 and NGC 1783, revealing elongated sequences in the ChM associated with metallicity variations. Finally, we investigate the 1G color distribution in 51 GCs, finding no connections with the host cluster parameters. These results shed light on the complex nature of 1G stars, providing insights into the GC formation environment. [ABSTRACT FROM AUTHOR]

About one-fifth of the Galactic globular clusters (GCs), dubbed Type II GCs, host distinct stellar populations with different heavy elements abundances. NGC 1851 is one of the most studied Type II GCs, surrounded by several controversies regarding the spatial distribution of its populations and the presence of star-to-star [Fe/H], C+N+O, and age differences. This paper provides a detailed characterization of its stellar populations through Hubble Space Telescope (HST), ground-based, and Gaia photometry. We identified two distinct populations with different abundances of s -process elements along the red-giant branch (RGB) and the subgiant branch (SGB) and detected two subpopulations among both s -poor (canonical) and s -rich (anomalous) stars. To constrain the chemical composition of these stellar populations, we compared observed and simulated colours of stars with different abundances of He, C, N, and O. It results that the anomalous population has a higher CNO overall abundance compared to the canonical population and that both host stars with different light-element abundances. No significant differences in radial segregation between canonical and anomalous stars are detected, while we find that among their subpopulations, the two most chemical extremes are more centrally concentrated. Anomalous and canonical stars show different 2D spatial distributions outside ∼3 arcmin, with the latter developing an elliptical shape and a stellar overdensity in the north-east direction. We confirm the presence of a stellar halo up to ∼80 arcmin with Gaia photometry, tagging 14 and 5 of its stars as canonical and anomalous, respectively, finding a lack of the latter in the south/south-east field. [ABSTRACT FROM AUTHOR]

Young and intermediate-age star clusters of both Magellanic Clouds exhibit complex colour–magnitude diagrams. In addition to the extended main-sequence turn-offs (eMSTOs), commonly observed in star clusters younger than ∼2 Gyr, the clusters younger than ∼800 Myr exhibit split main sequences (MSs). These comprise a blue MS, composed of stars with low rotation rates, and a red MS, which hosts fast-rotating stars. While it is widely accepted that stellar populations with different rotation rates are responsible for the eMSTOs and split MSs, their formation and evolution are still debated. A recent investigation of the ∼1.7-Gyr-old cluster NGC 1783 detected a group of eMSTO stars extremely dim in ultraviolet (UV) bands. Here, we use multiband Hubble Space Telescope photometry to investigate five star clusters younger than ∼200 Myr, including NGC 1805, NGC 1818, NGC 1850, and NGC 2164 in the Large Magellanic Cloud, and the Small Magellanic Cloud cluster NGC 330. We discover a group of bright MS stars in each cluster that are significantly dim in the F 225 W and F 275 W bands, similar to what is observed in NGC 1783. Our result suggests that UV-dim stars are common in young clusters. The evidence that most of them populate the blue MS indicates that they are slow rotators. As a by-product, we show that the star clusters NGC 1850 and BHRT 5b exhibit different proper motions, thus corroborating the evidence that they are not gravitationally bound. [ABSTRACT FROM AUTHOR]

We use images collected with the near-infrared camera (NIRCam) onboard the JWST and with the Hubble Space Telescope (HST) to investigate multiple populations at the bottom of the main sequence (MS) of 47 Tucanae. The m F 115 W versus  m F 115 W − m F 322 W 2 colour–magnitude diagram (CMD) from NIRCam shows that, below the knee, the MS stars span a wide colour range, where the majority of M-dwarfs exhibit blue colours, and a tail of stars are distributed towards the red. A similar pattern is observed from the m F 160 W versus  m F 110 W − m F 160 W CMD from HST , and multiple populations of M-dwarfs are also visible in the optical m F 606 W versus  m F 606 W − m F 814 W CMD. The NIRCam CMD shows a narrow sequence of faint MS stars with masses smaller than |$0.1\, \mathcal {M}_{\odot }$|⁠. We introduce a chromosome map of M-dwarfs that reveals an extended first population and three main groups of second-population stars. By combining isochrones and synthetic spectra with appropriate chemical composition, we simulate colours and magnitudes of different stellar populations in the NIRCam filters (at metallicities [Fe/H] = −1.5 and [Fe/H] = −0.75) and identify the photometric bands that provide the most efficient diagrams to investigate the multiple populations in globular clusters. Models are compared with the observed CMDs of 47 Tucanae to constrain M-dwarfs' chemical composition. Our analysis suggests that the oxygen range needed to reproduce the colours of first- and second-population M-dwarfs is similar to that inferred from spectroscopy of red giants, constraining the proposal that the chemical variations are due to mass transfer phenomena in proto-clusters. [ABSTRACT FROM AUTHOR]

The presence of differential reddening in the direction of Galactic globular clusters (GCs) has proven to be a serious limitation in the traditional colour–magnitude diagram (CMD) analysis. Here, we estimate local reddening variations in the direction of 56 Galactic GCs. To do that, we use the public catalogues derived as part of the Hubble Space Telescope UV Legacy Survey of Galactic GCs, which include photometry in the F 275 W ,  F 336 W ,  F 438 W ,  F 606 W , and F 814 W filters. We corrected photometry for differential reddening and found that for 21 out of 56 GCs the adopted correction procedure significantly improved the CMDs. Moreover, we measure the reddening law in the direction of these clusters, finding that R V exhibits a high level of variability within the Galaxy, ranging from ∼2.0 to ∼4.0. The updated values of R V have been used to improve the determination of local reddening variations and derive high-resolution reddening maps in the direction of the 21 highly-reddened targets within our sample. To compare the results of the different clusters, we compute the 68th percentile of the differential-reddening distribution, |$\sigma _{\Delta A_{\rm F814W}}$|⁠. This quantity ranges from 0.003 to 0.030 mag and exhibits a significant anticorrelation with the absolute module of the Galactic latitude and a strong correlation with the average reddening in the direction of each cluster. Therefore, highly-reddened GCs located in the proximity of the Galactic plane typically show higher differential-reddening variations across their field of view. [ABSTRACT FROM AUTHOR]

In the past few years, we have undertaken an extensive investigation of star clusters and their stellar populations in the Large and Small Magellanic Clouds (LMC and SMC) based on archival images collected with the Hubble Space Telescope. We present photometry and astrometry of stars in 101 fields observed with the Wide Field Channel of the Advanced Camera for Surveys and the Ultraviolet and Visual Channel and the Near-Infrared Channel of Wide Field Camera 3. These fields comprise 113 star clusters. We provide differential-reddening maps for those clusters with significant reddening variations across the field of view. We illustrate various scientific outcomes that arise from the early inspection of the photometric catalogs. In particular, we provide new insights into the extended main-sequence turnoff (eMSTO) phenomenon: (i) We detected eMSTOs in two clusters, KMHK361 and NGC265, which had no previous evidence of multiple populations. This finding corroborates the conclusion that the eMSTO is a widespread phenomenon among clusters younger than ~2 Gyr. (ii) The homogeneous color-magnitude diagrams (CMDs) of 19 LMC clusters reveal that the distribution of stars along the eMSTO depends on cluster age. (iii) We discovered a new feature along the eMSTO of NGC1783, which consists of a distinct group of stars on the red side of the eMSTO in CMDs composed of UV filters. Furthermore, we derived the proper motions of stars in the fields of view of clusters with multi-epoch images. Proper motions allowed us to separate the bulk of bright field stars from cluster members and investigate the internal kinematics of stellar populations in various LMC and SMC fields. As an example, we analyze the field around NGC346 to disentangle the motions of its stellar populations, including NGC364 and BS 90, young and pre-main-sequence stars in the star-forming region associated with NGC346, and young and old field stellar populations of the SMC. Based on these results and the fields around five additional clusters, we find that young SMC stars exhibit elongated proper-motion distributions that point toward the LMC, thus providing new evidence for a kinematic connection between the LMC and SMC. [ABSTRACT FROM AUTHOR]

Pseudo two-colour diagrams or Chromosome maps (ChM) indicate that NGC 2808 host five different stellar populations. The existing ChMs have been derived by the Hubble Space Telescope photometry, and comprise of stars in a small field of view around the cluster centre. To overcome these limitations, we built a ChM with U ,  B ,  I photometry from ground-based facilities that disentangle the multiple stellar populations of NGC 2808 over a wider field of view. We used spectra collected by GIRAFFE@VLT in a sample of 70 red giant branch and seven asymptotic giant branch (AGB) stars to infer the abundances of C, N, O, Al, Fe, and Ni, which combined with literature data for other elements (Li, Na, Mg, Si, Ca, Sc, Ti, Cr, and Mn), and together with both the classical and the new ground-based ChMs, provide the most complete chemical characterization of the stellar populations in NGC 2808 available to date. As typical of the multiple population phenomenon in globular clusters, the light elements vary from one stellar population to another; whereas the iron peak elements show negligible variation between the different populations (at a level of ≲0.10 dex). Our AGB stars are also characterized by the chemical variations associated with the presence of multiple populations, confirming that this phase of stellar evolution is affected by the phenomenon as well. Intriguingly, we detected one extreme O-poor AGB star (consistent with a high He abundance), challenging stellar evolution models that suggest that highly He-enriched stars should avoid the AGB phase and evolve as AGB-manqué star. [ABSTRACT FROM AUTHOR]

Hubble Space Telescope (HST) photometry is providing an extensive analysis of globular clusters (GCs). In particular, the pseudo-two-colour diagram dubbed 'chromosome map (ChM)' allowed to detect and characterize their multiple populations with unprecedented detail. The main limitation of these studies is the small field of view of HST , which makes it challenging to investigate some important aspects of the multiple populations, such as their spatial distributions and the internal kinematics in the outermost cluster regions. To overcome this limitation, we analyse state-of-art wide-field photometry of 43 GCs obtained from ground-based facilities. We derived high-resolution reddening maps and corrected the photometry for differential reddening when needed. We use photometry in the U, B , and I bands to introduce the Δ c U, B, I versus ΔB, I ChM of red-giant branch (RGB) and asymptotic-giant branch stars. We demonstrate that this ChM, which is built with wide-band ground-based photometry, is an efficient tool to identify first- and second-generation stars (1G and 2G) over a wide field of view. To illustrate its potential, we derive the radial distribution of multiple populations in NGC 288 and infer their chemical composition. We present the ChMs of RGB stars in 29 GCs and detect a significant degree of variety. The fraction of 1G and 2G stars, the number of subpopulations, and the extension of the ChMs significantly change from one cluster to another. Moreover, the metal-poor and metal-rich stars of Type II GCs define distinct sequences in the ChM. We confirm the presence of extended 1G sequences. [ABSTRACT FROM AUTHOR]

M 22 (NGC 6656) is a chemically complex globular cluster-like system reported to harbour heavy element abundance variations. However, the extent of these variations and the origin of this cluster is still debated. In this work, we investigate the chemical in-homogeneity of M 22 using differential line-by-line analysis of high-quality (R = 110 000, S/N  = 300 per pixel at 514 nm) VLT/UVES spectra of six carefully chosen red giant branch stars. By achieving abundance uncertainties as low as ∼0.01 dex (∼2 per cent), this high precision data validates the results of previous studies and reveals variations in Fe, Na, Si, Ca, Sc, Ti, Cr, Mn, Co, Ni, Zn, Y, Zr, La, Ce, Nd, Sm, and Eu. Additionally, we can confirm that the cluster hosts two stellar populations with a spread of at least 0.24 dex in [Fe/H] and an average s-process abundance spread of 0.65 dex. In addition to global variations across the cluster, we also find non-negligible variations within each of the two populations, with the more metal-poor population hosting larger spreads in elements heavier than Fe than the metal-rich. We address previous works that do not identify anomalous abundances and relate our findings to our current dynamical understanding of the cluster. Given our results, we suggest that M 22 is either a nuclear star cluster, the product of two merged clusters, or an original building block of the Milky Way. [ABSTRACT FROM AUTHOR]

Disentangling distinct stellar populations along the red-giant branches (RGBs) of globular clusters (GCs) is possible by using the pseudo-two-colour diagram dubbed chromosome map (ChM). One of the most intriguing findings is that the so-called first-generation (1G) stars, characterized by the same chemical composition of their natal cloud, exhibit extended sequences in the ChM. Unresolved binaries and internal variations in helium or metallicity have been suggested to explain this phenomenon. Here, we derive high-precision Hubble Space Telescope photometry of the GCs NGC 6362 and NGC 6838 and build their ChMs. We find that both 1G RGB and main-sequence (MS) stars exhibit wider ChM sequences than those of second-generation (2G). The evidence of this feature even among unevolved 1G MS stars indicates that chemical inhomogeneities are imprinted in the original gas. We introduce a pseudo-two-magnitude diagram to distinguish between helium and metallicity, and demonstrate that star-to-star metallicity variations are responsible for the extended 1G sequence. Conversely, binaries provide a minor contribution to the phenomenon. We estimate that the metallicity variations within 1G stars of 55 GCs range from less than [Fe/H]∼0.05 to ∼0.30 and mildly correlate with cluster mass. We exploit these findings to constrain the formation scenarios of multiple populations showing that they are qualitatively consistent with the occurrence of multiple generations. In contrast, the fact that 2G stars have more homogeneous iron content than the 1G challenges the scenarios based on accretion of material processed in massive 1G stars on to existing protostars. [ABSTRACT FROM AUTHOR]

The amount of mass lost by stars during the red-giant branch (RGB) phase is one of the main parameters to understand and correctly model the late stages of stellar evolution. Nevertheless, a fully comprehensive knowledge of the RGB mass-loss is still missing. Galactic Globular Clusters (GCs) are ideal targets to derive empirical formulations of mass-loss, but the presence of multiple populations with different chemical compositions has been a major challenge to constrain stellar masses and RGB mass-losses. Recent work has disentangled the distinct stellar populations along the RGB and the horizontal branch (HB) of 46 GCs, thus providing the possibility to estimate the RGB mass-loss of each stellar population. The mass-losses inferred for the stellar populations with pristine chemical composition (called first-generation or 1G stars) tightly correlate with cluster metallicity. This finding allows us to derive an empirical RGB mass-loss law for 1G stars. In this paper, we investigate seven GCs with no evidence of multiple populations and derive the RGB mass-loss by means of high-precision Hubble-Space Telescope photometry and accurate synthetic photometry. We find a cluster-to-cluster variation in the mass-loss ranging from ∼0.1 to ∼0.3 M⊙. The RGB mass-loss of simple-population GCs correlates with the metallicity of the host cluster. The discovery that simple-population GCs and 1G stars of multiple population GCs follow similar mass-loss versus metallicity relations suggests that the resulting mass-loss law is a standard outcome of stellar evolution. [ABSTRACT FROM AUTHOR]

Molecular‐based tools sometimes are the only laboratory techniques available to detect a recently discovered agent, and their validation without the existence of previously described 'gold standard' methods poses a challenge for the diagnosticians. A good example within this scenario is the recently described porcine circovirus 3 (PCV‐3) in the swine population worldwide, from which only few PCR methods have been described. Therefore, the primary objective of this study was to estimate the diagnostic accuracy of a direct PCR (dPCR) and a real‐time qPCR (qPCR) for detection of PCV‐3 in Italian swine population. Bayesian latent class analysis approach was used to rigorously assess their features and applicability in routine diagnostic activity. Data on dPCR and qPCR were available from 116 domestic pigs, which were randomly selected from 55 farms located at different regions in Northern Italy. The sensitivity (Se) estimates of dPCR (94%; posterior credible interval (PCI%) 84–100) and qPCR (96%; PCI% 90–100) were high and similar. The estimated specificity (Sp) of both dPCR and qPCR assays was around 97%. dPCR and qPCR assays showed a high and comparable Se and Sp estimates for the detection of PCV‐3 in Italian swine population. Significance and Impact of the Study: The continuous discovery of new pathogens poses a challenge in the development and evaluation of adequate diagnostic tools. In fact, since molecular‐based tools sometimes are the only available laboratory techniques, it is typically difficult to evaluate their diagnostic performances in the absence of a gold standard. The present study assesses this issue, demonstrating the excellent performances of two PCR‐based assays for porcine circovirus 3 (PCV‐3) detection using a Bayesian latent class analysis approach. Therefore, the molecular tests evaluated under this study constitute reliable tools for the routine diagnosis and surveillance programs of PCV‐3 circulating in swine populations. [ABSTRACT FROM AUTHOR]

Summary: Porcine circovirus 3 (PCV3) is a new species of the Circovirus genus, which has recently been associated with different clinical syndromes. Its presence has been reported in different countries of North and South America, Asia and recently also Europe (Poland). However, different from the other continents, no European PCV3 sequence is currently available in public databases. There is a strong need of epidemiological data and full‐genome sequences from Europe because of its relevance in the understanding of PCV3 molecular epidemiology and control. To fill this lack of information, samples collected in Denmark, Italy and Spain in 2016 and 2017 were screened for PCV3. Of the Danish samples, 36 of 38 the lymph nodes, six of 20 serum samples and two of 20 lung samples tested positive. Similarly, 10 of 29 lungs, 20 of 29 organ pools, six of 33 sera and one of eight nasal swabs tested PCV3 positive in Italy. Fourteen of 94 serum pools from seven of 14 Spanish farms were also positive. Despite the convenience nature of the sampling prevents any precise prevalence estimation, the preliminary screening of the data from three European countries confirmed a rather wide PCV3 distribution in Europe. Furthermore, the analysis of the six obtained complete European PCV3 genomes and their comparison with the public available sequences seems to support a remarkable worldwide PCV3 circulation. These results underline once more the urgency of more extensive epidemiological studies to refine the current knowledge on PCV3 evolution, transmission, spreading patterns and impact on pig health. [ABSTRACT FROM AUTHOR]

Research Highlights: Wild birds sampled in Italy tested for aMPV detection and characterization.aMPV-B found for the first time in a wintering Northern shoveler.Close phylogenetic relationship with aMPV-B strains circulating in Italian poultry.

Infectious bronchitis virus (IBV) is a pathogen causing respiratory, renal and reproductive clinical forms in chickens of all ages and productive categories. Its proneness to mutation and recombination gave rise to a plethora of variants differing in terms of pathogenicity, antigenicity, and distribution, with relevant implications for disease control, mainly pursued by routine vaccination, and diagnosis, requiring a steady update of molecular and serological methods. Among the most recent additions to the current phylogenetic classification, based on S1 gene sequencing, is the discovery of an eighth genotype (GVIII), further divided into lineages GVIII-1 and GVIII-2. GVIII-2, whose best-known representative is IB80, has been reported since 2015 in Europe, the Middle East and Asia. Most detections occurred in layers and breeders and were possibly associated to egg production drops. However, experimental reproduction of its pathogenicity could not be achieved. The significant genetic divergence of GVIII from other genogroups prevents its detection by many of the commonly applied biomolecular tests, hampering the understanding of its spread and impact. This study describes the validation of a GVIII-specific RT-PCR assay and its application to analyze samples collected from layer farms in Italy, where the presence of this genotype has never been investigated. The in-house assay proved highly reliable and allowed to establish the circulation of GVIII-2 in the country: between April and June 2024, 11 out of 84 flocks (13.1%) in 5 out of 24 farms (20.8%) tested positive. Phylogenetically, all Italian strains clustered together, whereas their identity with IB80 was 96.8-97.7%. Reproductive signs were reported in one farm and respiratory signs in another, whereas no clinical findings were recorded in the remaining positive cases. Although no definitive conclusions are possible on their prevalence and clinical relevance, the obtained results suggest that the presence of GVIII-2 strains in Italy is neither novel nor sporadic, highlighting the need to revise diagnostic approaches and shed light on the epidemiology of this novel lineage., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Legnardi, Poletto, Franzo, Harper, Bianco, Andolfatto, Blanco, Biarnés, Ramon, Cecchinato and Tucciarone.)

Infectious bursal disease (IBD) is among the most impactful immunosuppressive diseases of poultry. Its agent, infectious bursal disease virus (IBDV), is prone to both mutation and reassortment, resulting in a remarkable variability. Traditionally, IBDV characterization relies on antigenicity and pathogenicity assessment, but multiple phylogenetic classifications have been recently proposed, whose implementation in molecular surveys helps generating informative and standardized epidemiological data. In the present study, the Algerian IBDV scenario was assessed based on the novel classification guidelines by sequencing portions of both genome segments. Seventy pools of bursal samples were collected in 2022-2023 in 11 districts of Northern Algeria, mostly from broiler flocks. Out of 55 (78.6%) positive flocks, 40 (57.1%) were infected by field strains, which were characterized as very virulent strains (genotype A3B2) and phylogenetically related to previously reported Algerian strains. Significant differences in the percentage of field infections were observed between vaccinated (25/52, 46.2%) and unvaccinated (14/17, 82.3%) groups, and also between birds immunized with live intermediate (13/20, 65.0%) and intermediate plus (10/28, 35.7%) vaccines. Nonetheless, the number of field strain detections suggests a high infectious pressure and the inadequacy of current vaccination efforts, demanding a reevaluation of control measures coupled with attentive monitoring activities.

The clinical relevance of avian metapneumovirus (aMPV) is growing in the poultry sector, especially in broiler farming, where no vaccination is administered in Italy. Given the naïve status of the birds, a serological survey was conducted in a densely populated area of Northern Italy, to evaluate aMPV circulation. Seven farms were selected and sampled in summer/fall, then sampling was repeated in the following season (winter/spring) to assess a possible seasonal effect. In each farm, fifteen birds were blood sampled towards the end of the cycle and sera were analyzed with an ELISA test. Clinical signs were reported in 5 out of 7 farms, although all farms were positive at both sampling points, except for one, which was negative at the first sampling. The seroprevalence within farm ranged from 26.6% to 100%, and antibody titres appear to increase with age. No seasonality effect was evidenced, whereas a farm effect was more distinct. aMPV circulation appears wide in Northern Italian farms, with different clinical outcomes that could be modulated by intrinsic characteristics of the farms. In absence of vaccination, serological monitoring can be a useful tool for viral entrance monitoring, although sampling timing should be evaluated in order to spot seroconversion after late infections., Competing Interests: DISCLOSURES The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Introduction: Porcine circovirus 2 (PCV2) emerged more than three decades ago as one of the most impactful pathogens in the swine industry. Despite being a DNA virus, one of the hallmarks of PCV2 is its high evolutionary rate, which has led to the emergence of different genotypes, each exhibiting varying degrees of evolutionary success. Current knowledge suggests the occurrence of three main waves of genotype dominance, alternating over time (i.e., PCV2a, PCV2b, and PCV2d), alongside less prevalent genotypes. However, although PCV2d is currently the most common genotype nowadays, the others continue being circulating in the pig population., Methods: The present study reconsidered the epidemiological and evolutionary patterns of PCV2 genotypes using phylodynamic analyses, benefiting from an almost 10-fold increase in ORF2 sequence availability compared to previous studies. Additionally, a phylogeographic analysis was performed to investigate viral dispersal patterns and frequency, and the selective pressures acting on the capsid protein were estimated and compared among genotypes., Results: While successive emergence of major genotypes was confirmed, this study extends previous findings by revealing subsequent prevalence fluctuations of PCV2a and PCV2b after their initial decline. This evolutionary process may represent an example of balancing selection, specifically negative frequency-dependent selection, where a genotype fitness diminishes as it becomes more common, and vice versa. Variations in genotype- or clade-specific immunity-affected by the local prevalence of viral groups-combined with the periodic introduction of strains that have independently evolved in different regions, may have led to fluctuations in the population dynamics of major genotypes over time. These fluctuations were associated with ongoing evolution and variations in the capsid amino acid profile., Discussion: These findings have profound implications for future control strategies. Although PCV2d remains the most prevalent and widespread genotype, other genotypes should not be neglected. Control strategies should thus target the entire PCV2 population, with a focus on fostering broader and more cross-protective immunity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Franzo, Tucciarone, Legnardi, Drigo and Segalés.)

This case report presents the first molecular identification of a gammacoronavirus in a free-ranging striped dolphin ( Stenella coeruleoalba ) that was found stranded along the Croatian coastline in 2022. The dolphin exhibited a concurrent infection with cetacean morbillivirus. The gammacoronavirus strain was amplified and sequenced from heart tissue imprinted on an FTA ® card, revealing a notable genetic distance (approximately 8%) from previously characterized cetacean gammacoronaviruses. This finding highlights the importance of including gammacoronaviruses in routine diagnostics for stranded dolphins to gather epidemiological data on their prevalence and potential role in causing disease in cetaceans. This study sets the premises for a further understanding of the diversity and distribution of gammacoronaviruses in marine mammals and highlights the necessity for ongoing surveillance of emerging infectious diseases in wild populations.

Introduction: Porcine circovirus 2 (PCV-2) is a key pathogen for the swine industry at a global level. Nine genotypes, differing in epidemiology and potentially virulence, emerged over time, with PCV-2a, -2b, and -2d being the most widespread and clinically relevant. Conversely, the distribution of minor genotypes appears geographically and temporally restricted, suggesting lower virulence and different epidemiological drivers. In 2022, PCV-2e, the most genetically and phenotypically divergent genotype, was identified in multiple rural farms in North-eastern Italy. Since rural pigs often have access to outdoor environment, the introduction from wild boars was investigated., Methods: Through a molecular and spatial approach, this study investigated the epidemiology and genetic diversity of PCV-2 in 122 wild boars across different provinces of North-eastern Italy., Results: Molecular analysis revealed a high PCV-2 frequency (81.1%, 99/122), and classified the majority of strains as PCV-2d (96.3%, 78/81), with sporadic occurrences of PCV-2a (1.2%, 1/81) and PCV-2b (2.5%, 2/81) genotypes. A viral flow directed primarily from domestic pigs to wild boars was estimated by phylogenetic and phylodynamic analyses., Discussion: These findings attested that the genotype replacement so far described only in the Italian domestic swine sector occurred also in wild boars. and suggested that the current heterogeneity of PCV-2d strains in Italian wild boars likely depends more on different introduction events from the domestic population rather than the presence of independent evolutionary pressures. While this might suggest PCV-2 circulation in wild boars having a marginal impact in the industrial sector, the sharing of PCV-2d strains across distinct wild populations, in absence of a consistent geographical pattern, suggests a complex interplay between domestic and wild pig populations, emphasizing the importance of improved biosecurity measures to mitigate the risk of pathogen transmission., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Faustini, Poletto, Baston, Tucciarone, Legnardi, Dal Maso, Genna, Fiorentini, Di Donato, Perulli, Cecchinato, Drigo and Franzo.)

Avian metapneumovirus (aMPV) has been identified as an important cause of respiratory and reproductive disease, leading to significant productive losses worldwide. Different subtypes have been found to circulate in different regions, with aMPV-A and B posing a significant burden especially in the Old World, and aMPV-C in North America, albeit with limited exceptions of marginal economic relevance. Recently, both aMPV-A and aMPV-B have been reported in the U.S.; however, the route of introduction has not been investigated. In the present study, the potential importation pathways have been studied through phylogenetic and phylodynamic analyses based on a broad collection of partial attachment (G) protein sequences collected worldwide. aMPV-B circulating in the U.S. seems the descendant of Eastern Asian strains, which, in turn, are related to European ones. A likely introduction pathway mediated by wild bird migration through the Beringian crucible, where the East Asian and Pacific American flight paths intersect, appears likely and was previously reported for avian influenza. aMPV-A, on the other hand, showed a Mexican origin, involving strains related to Asian ones. Given the low likelihood of trade or illegal importation, the role of wild birds appears probable also in this case, since the region is covered by different flight paths directed in a North-South direction through America. Since the information on the role of wild birds in aMPV epidemiology is still scarce and scattered, considering the significant practical implications for the poultry industry demonstrated by recent U.S. outbreaks, further surveys on wild birds are encouraged.

Infectious bursal disease virus (IBDV) is a significant burden for poultry production and market due to both direct disease and induced immunosuppression. In the present study, the expression of different cytokines in the bursa of Fabricius and thymus was evaluated during a 28-day-long experimental infection with two strains classified in the G1a (Classical) and G6 (ITA) genogroups. Although both strains significantly affected and modulated the expression of different molecules, the G6 strain seemed to induce a delayed immune response or suppress it more promptly. A recovery in the expression of several mediators was observed in the G1a-infected group at the end of the study, but not in the G6 one, further supporting a more persistent immunosuppression. This evidence fits with the higher replication level previously reported for the G6 and with the clinical outcome, as this genotype, although subclinical, has often been considered more immunosuppressive. However, unlike other studies focused on shorter time periods after infection, the patterns observed in this paper were highly variable and complex, depending on the strain, tissue, and time point, and characterized by a non-negligible within-group variability. Besides confirming the strain/genogroup effect on immune system modulation, the present study suggests the usefulness of longer monitoring activities after experimental infection to better understand the complex patterns and interactions with the host response.

Avian reovirus (ARV) infection can cause significant losses to the poultry industry. Disease control has traditionally been attempted mainly through vaccination. However, the increase in clinical outbreaks in the last decades demonstrated the poor effectiveness of current vaccination approaches. The present study reconstructs the evolution and molecular epidemiology of different ARV genotypes using a phylodynamic approach, benefiting from a collection of more than one thousand sigma C (σC) sequences sampled over time at a worldwide level. ARVs' origin was estimated to occur several centuries ago, largely predating the first clinical reports. The origins of all genotypes were inferred at least one century ago, and their emergence and rise reflect the intensification of the poultry industry. The introduction of vaccinations had only limited and transitory effects on viral circulation and further expansion was observed, particularly after the 1990s, likely because of the limited immunity and the suboptimal and patchy vaccination application. In parallel, strong selective pressures acted with different strengths and directionalities among genotypes, leading to the emergence of new variants. While preventing the spread of new variants with different phenotypic features would be pivotal, a phylogeographic analysis revealed an intricate network of viral migrations occurring even over long distances and reflecting well-established socio-economic relationships.

The GI-19 lineage of infectious bronchitis virus (IBV) has emerged as one of the most impactful, particularly in the "Old World". Originating in China several decades ago, it has consistently spread and evolved, often forming independent clades in various areas and countries, each with distinct production systems and control strategies. This study leverages this scenario to explore how different environments may influence virus evolution. Through the analysis of the complete S1 sequence, four datasets were identified, comprising strains of monophyletic clades circulating in different continents or countries (e.g., Asia vs. Europe and China vs. Thailand), indicative of single introduction events and independent evolution. The population dynamics and evolutionary rate variation over time, as well as the presence and intensity of selective pressures, were estimated and compared across these datasets. Since the lineage origin (approximately in the mid-20th century), a more persistent and stable viral population was estimated in Asia and China, while in Europe and Thailand, a sharp increase following the introduction (i.e., 2005 and 2007, respectively) of GI-19 was observed, succeeded by a rapid decline. Although a greater number of sites on the S1 subunit were under diversifying selection in the Asian and Chinese datasets, more focused and stronger pressures were evident in both the European (positions 2, 52, 54, 222, and 379 and Thai (i.e., positions 10, 12, 32, 56, 62, 64, 65, 78, 95, 96, 119, 128, 140, 182, 292, 304, 320, and 323) strains, likely reflecting a more intense and uniform application of vaccines in these regions. This evidence, along with the analysis of control strategies implemented in different areas, suggests a strong link between effective, systematic vaccine implementation and infection control. However, while the overall evolutionary rate was estimated at approximately 10 -3 to 10 -4 , a significant inverse correlation was found between viral population size and the rate of viral evolution over time. Therefore, despite the stronger selective pressure imposed by vaccination, effectively constraining the former through adequate control strategies can efficiently prevent viral evolution and the emergence of vaccine-escaping variants.

Research Highlights: Different field IBDVs were found to circulate in the Near and Middle East.Multiple atypical genotypes (A3B1, A4B1, A6B1) were found to circulate extensively.Traditional very virulent IBDVs (A3B2) were a minority of the detected strains.Viral exchanges can be hypothesized between the region and different continents.

Fowl adenoviruses (FAdVs, species FAdV-A/-E) are responsible for several clinical syndromes reported with increasing frequency in poultry farms in the last decades. In the present study, a phylodynamic analysis was performed on a group of FAdV-D Hexon sequences with adequate available metadata. The obtained results demonstrated the long-term circulation of this species, at least several decades before the first identification of the disease. After a period of progressive increase, the viral population showed a high-level circulation from approximately the 1960s to the beginning of the new millennium, mirroring the expansion of intensive poultry production and animal trade. At the same time, strain migration occurred mainly from Europe to other continents, although other among-continent connections were estimated. Thereafter, the viral population declined progressively, likely due to the improved control measures, potentially including the development and application of FAdV vaccines. An increase in the viral evolutionary rate featured this phase. A role of vaccine-induced immunity in shaping viral evolution could thus be hypothesized. Accordingly, several sites of the Hexon, especially those targeted by the host response were proven under a significant pervasive or episodic diversifying selection. The present study results demonstrate the role of intensive poultry production and market globalization in the rise of FAdV. The applied control strategies, on the other hand, were effective in limiting viral circulation and shaping its evolution.

Stranded animals offer valuable information on marine mammal physiology and pathology; however, the decomposition state of the carcasses and lack of a rigorous cold chain for sample preservation can sometimes discourage diagnostic analyses based on nucleic acid detection. The present paper aims at evaluating the reliability of FTA ® card tissue imprints as an alternative matrix to frozen tissues for virological analyses based on biomolecular methods. Given the contribution of Cetacean morbillivirus (CeMV) to strandings and the increase of herpesvirus detection in cetaceans, these two pathogens were selected as representative of RNA and DNA viruses. Dolphin morbillivirus (DMV) and herpesvirus presence was investigated in parallel on tissue imprints on FTA ® cards and frozen tissues collected during necropsy of dolphins stranded in Italy. Samples were analysed by nested RT-PCR for DMV and nested-PCR for herpesvirus. Only one animal was positive for herpesvirus, hampering further considerations on this virus. DMV was detected in all animals, both in FTA ® card imprints and tissue samples, with differences possibly related to the decomposition condition category of the carcasses. Tissue sampling on FTA ® cards seems a promising alternative to frozen tissues for biomolecular analyses, especially when ensuring adequate storage and shipment conditions for frozen tissues is difficult.

Infectious bursal disease (IBD) is an immunosuppressive disease causing significant damage to the poultry industry worldwide. Its etiological agent is infectious bursal disease virus (IBDV), a highly resistant RNA virus whose genetic variability considerably affects disease manifestation, diagnosis and control, primarily pursued by vaccination. In Egypt, very virulent strains (genotype A3B2), responsible for typical IBD signs and lesions and high mortality, have historically prevailed. The present molecular survey, however, suggests that a major epidemiological shift might be occurring in the country. Out of twenty-four samples collected in twelve governorates in 2022-2023, seven tested positive for IBDV. Two of them were A3B2 strains related to other very virulent Egyptian isolates, whereas the remaining five were novel variant IBDVs (A2dB1b), reported for the first time outside of Eastern and Southern Asia. This emerging genotype spawned a large-scale epidemic in China during the 2010s, characterized by subclinical IBD with severe bursal atrophy and immunosuppression. Its spread to Egypt is even more alarming considering that, contrary to circulating IBDVs, the protection conferred by available commercial vaccines appears suboptimal. These findings are therefore crucial for guiding monitoring and control efforts and helping to track the spread of novel variant IBDVs, possibly limiting their impact.

Introduction: Porcine circovirus 3 (PCV-3) was firstly reported in 2017. Although evidence of its pathogenic role has been provided, its clinical relevance seems lower than Porcine circovirus 2 (PCV-2), as well as its evolutionary rate. Different studies have reported a high PCV-3 prevalence in wild boars, sometimes higher than the one observed in commercial pigs. Nevertheless, to date, few studies have objectively investigated the relationships between these populations when inhabiting the same area. Moreover, the role of small-scale, backyard pig production in PCV-3 epidemiology is still obscure., Methods: The present study investigated PCV-3 occurrence in 216 samples collected from the same area of Northern Italy from commercial and rural pigs, and wild boars. PCV-3 presence was tested by qPCR and complete genome or ORF2 sequences were obtained when possible and analysed using a combination of statistical, phylogenetic and phylodynamic approaches., Results: A higher infection risk in wild boars and rural pigs compared to the commercial ones was demonstrated. The phylodynamic analysis confirmed a larger viral population size in wild and rural populations and estimated a preferential viral flow from these populations to commercial pigs. A significant flow from wild to rural animals was also proven. The analysis of the Italian sequences and the comparison with a broader international reference dataset highlighted the circulation of a highly divergent clade in Italian rural pigs and wild boars only., Discussion: Overall, the present study results demonstrate the role of non-commercial pig populations in PCV-3 maintenance, epidemiology and evolution, which could represent a threat to intensive farming., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Franzo, Faustini, Legnardi, Berto, Dal Maso, Genna, Menandro, Poletto, Cecchinato, Drigo and Tucciarone.)

Porcine circovirus type 2 (PCV-2) is among the most burdensome viruses of the swine industry globally. Several genotypes have been periodically emerging, but just three of them (PCV-2a, PCV-2b, and PCV-2d) seem to circulate worldwide and be associated with the disease. Conversely, the spatial-temporal distribution of minor genotypes appears limited and their clinical relevance is still unclear. Recently PCV-2e was incidentally detected for the first time in Europe in a breeding farm in Northeastern Italy, while no connection could be established with countries where this genotype had been previously detected. To investigate circulating genotypes in the neglected rural context and provide a comparison with the most explored industrial context, a molecular survey was performed on samples collected in rural (n = 72) and industrial farms (n = 110) located in the same geographic area. Phylogenetic analysis surprisingly evidenced PCV-2e circulation only in pigs reared in backyard farms (n = 5), while major genotypes (PCV-2a, -2b, -2d) circulate in both rearing contexts. However, the close genetic similarity between the herein detected PCV-2e strains and the previously reported one testify that, although unusual, such rural-to-industrial strains exchange affected also PCV-2e. The greater genetic and phenotypic diversity of PCV-2e genotype compared to other ones might threaten the protection granted by current vaccines. The present study suggests the rural context as an ecological niche for the circulation of PCV-2e, and even of other minor genotypes. PCV-2e detection in pigs with outdoor access further stresses the epidemiological role of backyard farms as interfaces for pathogen introduction, potentially ascribable to the different rearing approaches, lower managerial and biosecurity capabilities, and easier contacts with wildlife., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

In future decades, the demand for poultry meat and eggs is predicted to considerably increase in pace with human population growth. Although this expansion clearly represents a remarkable opportunity for the sector, it conceals a multitude of challenges. Pollution and land erosion, competition for limited resources between animal and human nutrition, animal welfare concerns, limitations on the use of growth promoters and antimicrobial agents, and increasing risks and effects of animal infectious diseases and zoonoses are several topics that have received attention from authorities and the public. The increase in poultry production must be achieved mainly through optimization and increased efficiency. The increasing ability to generate large amounts of data ("big data") is pervasive in both modern society and the farming industry. Information accessibility-coupled with the availability of tools and computational power to store, share, integrate, and analyze data with automatic and flexible algorithms-offers an unprecedented opportunity to develop tools to maximize farm profitability, reduce socio-environmental impacts, and increase animal and human health and welfare. A detailed description of all topics and applications of big data analysis in poultry farming would be infeasible. Therefore, the present work briefly reviews the application of sensor technologies, such as optical, acoustic, and wearable sensors, as well as infrared thermal imaging and optical flow, to poultry farming. The principles and benefits of advanced statistical techniques, such as machine learning and deep learning, and their use in developing effective and reliable classification and prediction models to benefit the farming system, are also discussed. Finally, recent progress in pathogen genome sequencing and analysis is discussed, highlighting practical applications in epidemiological tracking, and reconstruction of microorganisms' population dynamics, evolution, and spread. The benefits of the objective evaluation of the effectiveness of applied control strategies are also considered. Although human-artificial intelligence collaborations in the livestock sector can be frightening because they require farmers and employees in the sector to adapt to new roles, challenges, and competencies-and because several unknowns, limitations, and open-ended questions are inevitable-their overall benefits appear to be far greater than their drawbacks. As more farms and companies connect to technology, artificial intelligence (AI) and sensing technologies will begin to play a greater role in identifying patterns and solutions to pressing problems in modern animal farming, thus providing remarkable production-based and commercial advantages. Moreover, the combination of diverse sources and types of data will also become fundamental for the development of predictive models able to anticipate, rather than merely detect, disease occurrence. The increasing availability of sensors, infrastructures, and tools for big data collection, storage, sharing, and analysis-together with the use of open standards and integration with pathogen molecular epidemiology-have the potential to address the major challenge of producing higher-quality, more healthful food on a larger scale in a more sustainable manner, thereby protecting ecosystems, preserving natural resources, and improving animal and human welfare and health.

Peste des petits ruminants (PPR) is a burdensome viral disease primarily affecting small ruminants, which is currently targeted for eradication by 2030 through the implementation of a Global Control and Eradication Strategy (PPR GCES). The PPR GCES, launched in 2015, has strongly encouraged countries to participate in Regional PPR Roadmaps, designated according to the Food and Agricultural Organization of the United Nations (FAO) and World Organisation for Animal Health (WOAH) regions and epidemiological considerations, with each targeted by dedicated meetings and activities. Following the conclusion of the first phase of the PPR Global Eradication Program (PPR GEP) (2017-2021), the present work focuses on the disease situation and status of the eradication campaign in the fourteen countries of the PPR GCES Middle Eastern Roadmap as well as Egypt. PPR is endemic to or suspected to be present in most of the region, except for Bahrain, which, as of 2021, is preparing to apply for official recognition as being free of PPR. Some substantial shortcomings are observed in surveillance and disease reporting, as well as in the implemented control strategies, most notably vaccination. Since many of these limitations are shared by many of the investigated countries, the international cooperation and harmonization of control efforts appears crucial to making PPR eradication attainable in the Middle East.