Avian Poxvirus Research Articles

Trichomonad Disease in Wild Turkeys (Meleagris gallopavo): Pathology and Molecular Characterization of Histomonas, Tetratrichomonas, Tritrichomonas, and Simplicimonas spp.

The Wild Turkey (Meleagris gallopavo) is a geographically widespread, popular game bird and conservation icon in North America. Following successful reestablishment in parts of the US and Canada, regional declines have fueled population health concerns; therefore, understanding mortality causes and spatiotemporal patterns of disease is important to uncover potential ongoing and future health risks. Histomonosis, caused by the trichomonad Histomonas meleagridis, is a well-established and potentially fatal disease in Wild Turkeys; however, its prevalence and potential population health impacts remain poorly understood. Moreover, molecular tools recently have allowed for the detection of additional trichomonads that similarly can cause fatal disease in Wild Turkeys. We describe and compare disease due to H. meleagridis with that of Tetratrichomonas gallinarum, Tritrichomonas sp., and Simplicimonas sp. in Wild Turkeys in the southeastern US. Among 857 Wild Turkeys evaluated postmortem from 2002 to 2023, 34 (4.0%) were diagnosed with trichomonad disease, often assumed to be histomonosis prior to molecular testing. However, among 25 debilitating to fatal trichomonad disease cases for which etiologies were confirmed by PCR from 2015 to 2023, H. meleagridis was detected in 16/25 (64.0%), T. gallinarum in 6/25 (24.0%), Tritrichomonas sp. in 2/25 (8.0%) and Simplicimonas sp. in 1/25 (4.0%). These turkeys had similar clinical manifestations, and although lesion patterns varied to some extent, liver and/or intestinal tract was most commonly affected. Coinfections were common among all turkeys with trichomonad disease from 2015 to 2023 (21/25, 84.0%) and included viruses (lymphoproliferative disease virus, avian poxvirus), bacteria (Streptococcus gallolyticus, Listeria monocytogenes, Escherichia coli), and other protozoa (Sarcocystis sp., Haemoproteus sp.). Our results highlight the importance of molecular diagnostic testing in determining etiologies of trichomonad disease in Wild Turkeys. Further evaluation of the epidemiology and pathogenesis of trichomonad disease and its varied etiologies in Wild Turkeys and other birds is warranted to better understand risk factors and potential health impacts.

Determination of non-cytotoxic antiviral concentrations of purine and indole derivatives in vitro

Mammalian cells from lines derived from bovine embryonic trachea (EBTr) were used in the present study. After the formation of semi-confluent monolayers, one subpopulation of cells was inoculated with the vaccine avian poxvirus strain FK (fowl) and the other with the vaccine avian poxvirus strain Dessau (pigeon) (Poxviridae family). Twenty-four hours after viral inoculation, individual subsets of infected cells were treated with the purine derivative aminophylline and the remaining subsets with the indole derivative ergotamine tartrate, respectively. The effects of the analogues thus administered on the cells were recorded at the 24th hour and 48th hour after treatment with each substance, respectively. In cells inoculated with the FK viral strain and treated with both aminophylline and ergotamine tartrate, decreased cell viability was observed at all dilutions at the 48th hour post-treatment compared to the 24th hour. In aminophylline-treated cells, these differences were not statistically significant, unlike in ergotamine tartrate-treated cells, where they were statistically significant. In the same cells infected with the Dessau strain and treated with aminophylline, although decreased cell viability was found at the 48th hour of treatment compared to the 24th hour, in most cases no statistically significant differences were found. In cells infected with the same strain but treated with ergotamine tartrate, despite the lack of statistically significant differences, increased cell viability was seen at the 48th hour post-treatment compared to the 24th hour, specifically at the higher concentrations of 10-3 and 10-4 M/mL. These results suggest a lower cytotoxicity of aminophylline compared to ergotamine tartrate, but on the other hand, a higher anti-viral activity of ergotamine tartrate against the Dessau virus strain compared to aminophylline in in vitro conditions. Further studies need to be conducted in this regard.

Molecular Detection of Avian Poxvirus in Chickens and Pigeons of Diyala Province in Iraq

Fowlpox is an infectious viral disease affecting domestic and wild birds. The current study was conducted using PCR-based molecular detection to determine the avian pox virus (APV) and its genetic sequence. A total of 40 pigeons and 40 chickens suspected of APV were obtained from different bird markets. Following euthanasia, tissue samples were taken from the vents, eyes, wings, and beaks for molecular analysis. DNA extraction was focused on the core protein gene region (pb4). PCR results confirmed the presence of APV in all samples, generating specific cDNA bands of 329 bp. Four GenBank accession codes (OR670580, OR670581, OR670582, and OR670583) represented pigeon pox samples, whereas two GenBank accession numbers (OR619724 and OR619725) represented samples of fowlpox. A significant similarity of 99% was found by genetic analysis between the reference target sequences and the sequenced samples. In viral samples of fowlpox and avipoxvirus, nucleic acid variants (205 C > G and 204 T > C) were found with missense and silent effects on particular proteins (p.101Leu>Val and p.108Pro). Phylogenetic analysis organized the samples into clades representing fowlpox and pigeon pox viruses, showing close relationships with strains from different geographical regions. This study unequivocally demonstrates the susceptibility of both domestic and wild birds to avian pox, highlighting the pivotal role of phylogenetic analysis and molecular detection in elucidating novel perspectives on the genetic landscape of these viruses. Keywords: Fowl pox, Gene, Pigeon pox, Polymerase Chain Reaction

Systemic avian poxvirus infections associated with the B1 subclade of canarypox virus.

Avian poxvirus infections typically manifest as 2 forms: cutaneous ("dry") pox, characterized by proliferative nodules on the skin, and diphtheritic ("wet") pox, characterized by plaques of caseous exudate in the oropharynx and upper respiratory and gastrointestinal tracts. Systemic spread of virus to visceral organs beyond the skin and mucous membranes is rarely reported. Out of 151 cases diagnosed with avian poxvirus over a 20-year period at a zoological institution, 22 were characterized as having systemic involvement based on histopathology and molecular findings. Gross lesions in systemic cases included soft white nodules scattered throughout the liver, spleen, and kidneys. Two histopathologic patterns emerged: (1) widespread histiocytic inflammation in visceral organs with intrahistiocytic viral inclusions and (2) severe, localized dry or wet pox lesions with poxvirus-like inclusions within dermal and subepithelial histiocytes. In situ hybridization targeting the core P4b protein gene confirmed the presence of poxvirus DNA within histiocytes in both patterns. Polymerase chain reaction was performed targeting the reticuloendothelial virus long terminal repeat (REV LTR) flanking region and the core P4b protein gene. Sequences of the REV LTR flanking region from all systemic pox cases were identical to a previously described condorpox virus isolated from an Andean condor with systemic pox. Sequences of the core P4b protein gene from all systemic pox cases grouped into cluster 2 of the B1 subclade of canarypox viruses. Systemic involvement of avian poxvirus likely occurs as a result of infection with certain strain variations in combination with various possible host and environmental factors.

Genetic characteristics and pathogenicity of avian pox virus for a new host, Cherry Valley breeder ducks in China

ABSTRACT Cherry Valley breeder ducks in Shandong province in northern China have experienced swollen eyes, lachrymation, pox scabs on glabrous or glabrous skin, depression and dysentery since 2021. The spread of this infectious disease has seriously affected the breeder ducks in major Cherry Valley duck farms in China. The virus causing clinical signs in Cherry Valley breeder ducks was isolated by chicken embryo inoculation. We successfully isolated a strain of duck pox virus from diseased breeder ducks by virus replication. We have also successfully conducted experiments for duck pox disease using the isolated strains to infect ducklings. By comparison with 22 pox viruses already published in GenBank, the virus strain obtained in this study was most homologous (about 99.7%) to the strain isolated from infected domestic ducks in Guangxi, China in 2014 (KJ192189), and belonged to the same A5 subtype. Since there were no previous cases of avian pox virus infecting white or Cherry Valley duck breeds, this study identified a new host for avian pox virus infection and provided theoretical support and data for the development of avian pox virus research. RESEARCH HIGHLIGHTS Avian pox virus can infect a new host type – Cherry Valley breeder ducks. The avian pox virus isolated from Cherry Valley breeder ducks showed highest identity with Guangxi hemp duck-derived avian pox virus. Cherry Valley breeder ducks were infected with avian pox virus of subtype A5.

Recombination of Goose Parvovirus VP3 Gene and Goose Interferon Ɣ Gene from Fowlpox Virus Immune Protection and Its Mechanism

Gosling plague (GP) is caused by goose parvovirus (GPV) gosling acute, subacute, and septic infectious diseases, commonly known as gosling plague. At present, the prevention and control of GP at home and abroad mainly adopts the method of immunizing female geese with attenuated vaccine. Goslings can be immunized, but there are still the phenomenon of dispersive virus and virulence reverse. It, therefore, is urgent to develop a new safe and effective vaccine. The recombinant avian pox virus vaccines rFPV-GoIFNγ, rFPV-GPV-VP3 and rFPV-GoIFNγ-VP3 were used to monitor their antibody levels and evaluate the protective mechanism of the vaccine in this study. This study lays a foundation for further in vivo testing of recombinant avian pox virus vaccine for goose parvovirus and provides a reference for the development of recombinant avian pox virus vaccine for other avian infectious diseases.

Transcriptomic responses of Galápagos finches to avian poxvirus infection.

Emerging pathogens can have devastating effects on naïve hosts, but disease outcomes often vary among host species. Comparing the cellular response of different hosts to infection can provide insight into mechanisms of host defence. Here, we used RNA-seq to characterize the transcriptomic response of Darwin's finches to avian poxvirus, a disease of concern in the Galápagos Islands. We tested whether gene expression differs between infected and uninfected birds, and whether transcriptomic differences were related either to known antiviral mechanisms and/or the co-option of the host cellular environment by the virus. We compared two species, the medium ground finch (Geospiza fortis) and the vegetarian finch (Platyspiza crassirostris), to determine whether endemic Galápagos species differ in their response to pox. We found that medium ground finches had a strong transcriptomic response to infection, upregulating genes involved in the innate immune response including interferon production, inflammation, and other immune signalling pathways. In contrast, vegetarian finches had a more limited response, and some changes in this species were consistent with viral manipulation of the host's cellular function and metabolism. Many of the transcriptomic changes mirrored responses documented in model and in vitro studies of poxviruses. Our results thus indicate that many pathways of host defence against poxviruses are conserved among vertebrates and present even in hosts without a long evolutionary history with the virus. At the same time, the differences we observed between closely related species suggests that some endemic species of Galápagos finch could be more susceptible to avian pox than others.

Evaluation of minimally invasive sampling methods for detecting Avipoxvirus: Hummingbirds as a case example.

Avian pox is a common avian virus that in its cutaneous form can cause characteristic lesions on a bird's dermal surfaces. Detection of avian pox in free-ranging birds historically relied on observations of visual lesions and/or histopathology, both which can underestimate avian pox prevalence. We compared traditional visual observation methods for avian pox with molecular methods that utilize minimally invasive samples (blood, toenail clipping, feathers, and dermal swabs) in an ecologically important group of birds, hummingbirds. Specifically, avian pox prevalence in several species of hummingbirds were examined across multiple locations using three different methods: (1) visual inspection of hummingbirds for pox-like lesions from a long-term banding data set, (2) qPCR assay of samples from hummingbird carcasses from wildlife rehabilitation centers, and (3) qPCR assay of samples from live-caught hummingbirds. A stark difference in prevalences among these three methods was identified, with an avian pox prevalence of 1.5% from banding data, 20.4% from hummingbird carcasses, and 32.5% from live-caught hummingbirds in California. This difference in detection rates underlines the necessity of a molecular method to survey for avian pox, and this study establishes one such method that could be applied to other wild bird species. Across all three methods, Anna's hummingbirds harbored significantly higher avian pox prevalence than other species examined, as did males compared with females and birds caught in Southern California compared with Northern California. After hatch-year hummingbirds also harbored higher avian pox prevalences than hatch-year hummingbirds in the California banding data set and the carcass data set. This is the first study to estimate the prevalence of avian pox in hummingbirds and address the ecology of this hummingbird-specific strain of avian pox virus, providing vital information to inform future studies on this charismatic and ecologically important group of birds.

An Investigation of the Cause of Wild Turkey Mortality in Mississippi.

Lymphoproliferative disease virus (LPDV) is an exogenous alpharetrovirus that sporadically causes fatal lymphoid neoplasia in affected turkeys. Previous studies of wild turkeys (Meleagridis gallopavo) in the United States have demonstrated geographically widespread LPDV infection and frequent coinfection with avian poxvirus (APV) and reticuloendotheliosis virus (REV). This study was conducted to better understand health risks to Mississippi wild turkeys, including the relative importance of LPDV, APV, and REV in contributing to mortality. Thirteen wild turkeys, which died naturally or were euthanized due to illness, were submitted to Mississippi State University's Poultry Research and Diagnostic Laboratory for postmortem examinations. Birds originated from nine counties across the state over the past 5 yr. Carcasses were submitted as fresh (nonfrozen) or frozen. At autopsy, 9 of 13 turkeys had severe, proliferative cutaneous lesions on the head and neck, with diphtheritic or proliferative oral and esophageal lesions. Samples were collected for molecular diagnostic testing (LPDV and REV PCR), histopathology, and bacterial culture and isolation. External and internal parasites were preserved in formalin for identification. APV (cutaneous and/or diphtheritic forms) was diagnosed in 9 of 13 birds by identification of pathognomonic histologic lesions (including intracytoplasmic inclusion bodies). Interestingly, all birds with APV were also REV PCR positive. Furthermore, eight turkeys were positive for LPDV, and LPDV was commonly associated with coinfections with APV and REV.

Effects of urban sugar water feeding on bird body condition and avian diseases

Garden bird sugar water feeding is increasingly popular worldwide, but little is known about its effects on bird health and associated diseases. There is a concern that feeding stations can accumulate pathogens and facilitate pathogen transmission between individuals, resulting in adverse effects on body condition of visiting birds. We tested the effects of sugar water feeding in urban New Zealand backyards by sampling target species for multiple infections and comparing bird body condition. For this, we compared backyards with and without sugar water feeders and again compared existing sugar water feeders with various sugar concentrations in two cities and in two seasons. Birds caught in gardens with sugar water feeders had poorer body condition; however, birds had better body condition in the city with the warmer climate (Auckland), during summer, and in gardens with high (≥20%) sugar concentration in sugar water feeders in winter. All screening tests for Chlamydia psittaci and Salmonella spp. returned negative results. Avian poxvirus prevalence in tauhou ( Zosterops lateralis) was four times higher in the city with a warmer climate. The likelihood of lice infection in tauhou was lower in gardens with feeders, in the warmer city, in summer, and at feeders with higher sugar concentrations. In tūī ( Prosthemadera novaeseelandiae), the likelihood of lice infection decreased with an increase in sugar concentration. Coccidia infection was 4.25 times higher in tauhou in gardens with feeders. Despite the identified risks associated with sugar water feeding, there appear to be potential benefits for native nectarivorous birds, specifically in winter.

Microbiome composition of Anna's hummingbirds differs among regions of the gastrointestinal tract

The gastrointestinal (GI) microbiome is an important mediator and indicator of host physiology, health and fitness in many vertebrate systems, but sources of variation in microbiome composition within species are poorly understood, particularly in free‐ranging birds. Hummingbirds have GI anatomy and physiology similar to other avian species, but they have a relatively rapid gastrointestinal transit rate, which could affect microbiome structure. To date, it has not been elucidated if microbiomes isolated from feces and samples from sections of the GI tract differ in an avian organism that has a rapid gastrointestinal time. Here, we test the hypothesis that the microbiome in free‐ranging and wildlife rehabilitation‐sourced Anna's hummingbirds Calypte anna differ between fecal and samples from three sections of the GI tract and between birds that differ in sex, age and poxvirus infection status. We characterized bacterial composition in fecal and GI tract samples from Anna's hummingbirds by amplicon sequencing of the 16S rRNA gene. We found strong evidence of differentiation in bacterial composition among GI tract regions and compared it to fecal samples. Actinobacteria, primarily genus Corynebacterium, Firmicutes and Fusobacteria were abundant in bird samples from regions of the GI tract. In contrast, fecal bacterial communities were more diverse and variable compared to GI tract samples. Bacterial community composition differed between male and female hummingbirds and with bird age (hatch year vs after‐hatch year). Finally, birds with symptoms of avian poxvirus infection had a higher relative abundance of Staphylococcus spp. than birds with no symptoms of pox. Our results suggest that Anna's hummingbirds host differentiated microbiome among GI tract regions that is consistent among individual birds. The GI bacterial community also contained taxa not represented in fecal samples. This provides evidence for the possibility of a residential gut microbiome in Anna's hummingbirds, although functional significance of the bacterial microbiome remains unknown.

In ovo inhibition of avian pox virus replication by mangosteen rind and red ginger ethanolic extracts.

Background and Aim:Avian pox is a contagious disease caused by the avian pox virus (APV). Mangostin and γ-mangostin in mangosteen rind (MR) and gingerol in red ginger (RG) exhibit antiviral activity. In this study, we evaluated the effect of MR and RG ethanolic extracts on APV based on pock lesions on the chorioallantoic membrane (CAM) of specific pathogen-free (SPF) embryonated chicken eggs (ECEs).Materials and Methods:Three APVs from chicken isolates (C1, C2, and C3), one APV from a pigeon isolate (P), 1.5% and 3% MR ethanolic extract, 5% and 10% RG ethanolic extract, and a combination of 1.5% MR and 5% RG at 0.1 mL/egg were inoculated in ovo (7th day incubation, chorioallantoic route) in SPF ECEs. A control group inoculated in ovo with APV alone was also established. Each treatment consisted of three replicates. Parameters including embryo survival, CAM lesions, and average number of pock lesions were determined.Results:In ovo inoculation of MR and RG ethanolic extracts was not harmful to the ECEs and did not induce CAM lesions. The average number of pock lesions in the control group (C1, C2, C3, and P) was 35, 14, 10, and 17, respectively, whereas in all treatment groups, the number was 0, except in the 5% RG group of C1, which had a value of 10.Conclusion:In ovo inoculation of 1.5% and 3% MR, 5% and 10% RG, and the combination of 1.5% MR plus 5% RG ethanolic extracts at 0.1 mL/egg inhibit APV by reducing the number of pock lesions on the CAM of the ECE.

Genetic Sequencing of Attwater's Prairie Chicken Avian Poxvirus and Evaluation of Its Potential Role in Reticuloendotheliosis Virus Outbreaks.

Efforts to breed Attwater's prairie chickens (APC; Tympanuchus cupido attwateri) in captivity to supplement wild populations of this endangered bird have been negatively affected by infections with Avipoxvirus and reticuloendotheliosis virus (REV). Because REV can be integrated into the genome of fowlpox virus (FPV) and may be transmitted in that manner, identifying the source of avipox disease in APC is important to mitigate the impact of this virus. Tissue samples from APC were collected from breeding programs in Texas from 2016 to 2020. These samples consisted of 11 skin lesions and three internal organs from a total of 14 different birds that died of unknown causes or were euthanized. Avipoxvirus was detected by PCR and isolation in embryonating chicken eggs in all skin lesion samples but was not detected in internal organs. Using sequence analysis of FPV polymerase and 4b genes, we determined that 10 out of 11 Avipoxvirus detections resided within the fowlpox clade and a single sample resided within the canarypox clade. REV sequences were detected in all FPV positive samples and in all internal organ tissues but were not detected in the sample matching the canarypox clade. Analysis of REV sequences and PCR detection showed the REV infecting APC was consistent with REV-A and had little variability on analysis of the U3 region of the long terminal repeat. The results of this study indicate control of REV in APC breeding colonies may benefit by a vaccination program targeting FPV and REV. However, a commercially available vaccine for REV is not available at this time.

A Review on the Prevalence of Poxvirus Disease in Free-Living and Captive Wild Birds

Avian pox is a widespread infection in birds caused by genus Avipoxvirus pathogens. It is a noteworthy, potentially lethal disease to wild and domestic hosts. It can produce two different conditions: cutaneous pox, and diphtheritic pox. Here, we carry out an exhaustive review of all cases of avian pox reported from wild birds to analyze the effect and distribution in different avian species. Avian poxvirus strains have been detected in at least 374 wild bird species, a 60% increase on a 1999 review on avian pox hosts. We also analyze epizootic cases and if this disease contributes to wild bird population declines. We frequently observe very high prevalence in wild birds in remote island groups, e.g., Hawaii, Galapagos, etc., representing a major risk for the conservation of their unique endemic avifauna. However, the difference in prevalence between islands and continents is not significant given the few available studies. Morbidity and mortality can also be very high in captive birds, due to high population densities. However, despite the importance of the disease, the current detection rate of new Avipoxvirus strains suggests that diversity is incomplete for this group, and more research is needed to clarify its real extent, particularly in wild birds.

Detection of Laryngotracheitis Virus in Poultry Flocks with Respiratory Disorders in Slovenia.

Infectious laryngotracheitis (ILT) is an acute, highly contagious infectious disease of the upper respiratory tract in chickens and other poultry species that causes significant economic losses in countries worldwide. Between 2017 and 2019, seven outbreaks of mild to severe respiratory disorders with high suspicion of ILT occurred in commercial and backyard poultry flocks in Slovenia. In all submissions, infection with ILT virus (ILTV) was confirmed by PCR, which is the first report of ILT in Slovenia. Circulating ILT strains were characterized by the sequence and phylogenetic analysis of two fragments of the ICP4 gene. Four strains—three detected in non-vaccinated flocks and one in a flock vaccinated against ILT—were identical or very similar to the chicken embryo–origin live virus vaccines, and the other three were closely related to Russian, Chinese, Australian, and American field strains and to tissue culture origin vaccine strains. As in other diseases, coinfections with other respiratory pathogens in confirmed ILT cases may cause a more severe condition and prolong the course of the disease. In our study, coinfections with Mycoplasma synoviae (7/7 tested flocks), infectious bronchitis virus (5/5 tested flocks), Mycoplasma gallisepticum (4/7 tested flocks), Ornithobacterium rhinotracheale (3/4 tested flocks), and avian pox virus (1/2 tested flocks) were confirmed, indicating the importance of these pathogens in the occurrence of ILT infections.

A Case Report of Outbreak Avian Pox Virus from Layer Chickens and a Pigeon in Yogyakarta, Indonesia

A Case Report of Outbreak Avian Pox Virus from Layer Chickens and a Pigeon in Yogyakarta, Indonesia

Outbreaks of Avian Poxvirus Infections in Cormorants in Yamanashi Prefecture during 2017 and 2019

Outbreaks of Avian Poxvirus Infections in Cormorants in Yamanashi Prefecture during 2017 and 2019

Plasmodium relictum

Plasmodium relictum is a widespread haemosporidian parasite infecting over 300 bird species from all continents except Antarctica. Based on sequences of the cytochrome b gene, that has become the barcoding region for avian haemosporidians, five different mitochondrial haplotypes (lineages) have been linked to P. relictum (SGS1, GRW04, GRW11, LZFUS01, and PHCOL01). Culex mosquitoes are the main vectors in P. relictum transmission, while other potential vectors include Aedes, Lutzia, Culiseta, and Anopheles species. Introduction of the lineage GRW04 to Hawaii in the first half of the 20th century, where Culex vectors were previously introduced – in synergy with anthropogenic impacts and infections of other pathogens (i.e., avian pox virus) – resulted in dramatic population declines of native bird species. As a result of this and other invasion events, P. relictum is nowadays catalogued as one of the 100 world’s worst invasive species.

First data on breeding success of Croatian inland colonies of Common TernSterna hirundo

AbstractIn 2018 and 2019, the breeding success of two Common Tern colonies on artificial lakes near the River Sava in Zagreb, Croatia, was studied. The colonies were visited weekly from May to July and we collected data on phenology, number of breeding pairs, clutch size as well as egg and chick survival. We also conducted a comparison between early and late breeders. Hatching and fledging success was within previously observed ranges, apart from a low hatching success on Siromaja in 2019. The smaller colony on Siromaja had a higher productivity in both years than the colony on Rakitje, although in 2018 an avian pox virus killed much of the late chicks on Rakitje. Early breeders seem to have had higher hatching success and average clutch size. Furthermore, a greater proportion of them managed to hatch all their eggs compared to late breeders, but the differences were not statistically significant. Our study provided baseline data for future monitoring of phenology and breeding success with regard to the management of breeding colonies.

Avian Poxvirus Infection in Polish Great Tits (Parus Major).

IntroductionAvian poxvirus infections are widespread in the domestic poultry population but are also reported in wild birds. In poultry, these infections cause significant economic losses, while wild birds may be a reservoir for poxvirus which affects breeding poultry. However, wild birds may also exhibit characteristic anatomopathological changes. This study concerns the infection of wild-living great tits (Parus major) with the avian poxvirus in Poland.Material and MethodsSamples of internal organs and skin collected from great tits were homogenised and total cellular DNA was isolated. In PCR, the primers complementary to gene encoding the core protein 4b of the HP44 strain of fowl poxvirus (FPV) were used.ResultsAfter electrophoresis in 2% agarose gel, the PCR product of 578 bp characteristic for FPV was obtained in DNA samples isolated from skin lesions and the heart. The analysis of the nucleotide sequence of the virus strain showed 99% similarity to many poxviruses previously isolated from great tits and other free birds at various sites in the world.ConclusionsThis paper is the first clinically documented evidence obtained in laboratory conditions of avian poxvirus cases in great tits in Poland.