Circulation of Antibiotic-Resistant Escherichia coli in Wild and Domestic Waterfowl in Ukraine.

Background: Antibiotic resistance is becoming an increasingly urgent problem for human and animal health due to the widespread use of antibiotics in medicine, veterinary medicine, and agriculture. At the same time, the natural reservoirs of antibiotic-resistant pathogens remain unclear. Wild birds may play a role in this due to their biology. Escherichia coli is a representative indicator pathogen for antibiotic resistance studies. Materials and Methods: In 2020-2021, sampling of feces and cloacal swabs from six species of wild waterfowl (Eurasian wigeon Anas penelope, Eurasian teal Anas crecca, white-fronted goose Anser albifrons, red-breasted goose Rufibrenta ruficollis, graylag goose Anser anser, shelduck Tadorna tadorna) and from two species of domestic waterfowl (ducks and geese) was conducted in the Kherson, Zaporizhzhia, Odesa, Kharkiv, and Cherkasy regions of Ukraine. Biological material was collected, stored, and transported in cryotubes with transport medium (brain heart infusion broth [BHIB] with the addition of 15% glycerol) in liquid nitrogen. Bacteriological studies were carried out according to standard methods for the isolation and identification of microorganisms. Drug resistance of E. coli was carried out by a standard disk diffusion method. Results: Bacteria representing six families (Enterobacteriaceae, Yersiniaceae, Morganellaceae, Bacillaceae, Pseudomonadaceae, Staphylococcaceae) were isolated from clinically healthy wild birds (wigeon, Eurasian teal, white-fronted goose, red-breasted goose, mallard, graylag goose, shelduck) in the southern regions of Ukraine with isolation rates ranging from 26.7% to 100%. A total of 19 E. coli isolates were cultured from 111 samples from wild birds, and 30 isolates of E. coli were cultured from 32 poultry samples. E. coli was isolated from birds of all species. The prevalence of E. coli ranged from 5.0% to 33.3% in wild waterfowl and from 90.9% to 100% in domestic waterfowl. The prevalence of multidrug-resistant (MDR) E. coli ranged from 10.0% to 31.8% in wild and domestic waterfowl: 3 of 15 (20%) specimens from wild mallard were MDR in the Kherson region, as well as 7 of 22 domestic ducks (31.8%) and 1 of 10 geese (10%) in the Kharkiv and Cherkasy regions. Isolates from wild birds were the most resistant to ampicillin (AMP), amoxiclav (AMC), amoxicillin (AMX), doxycycline (DO), and chloramphenicol (C). Isolates from poultry were resistant to ampicillin, amoxiclav, doxycycline, amoxicillin, chloramphenicol, and enrofloxacin (EX). Most of the other E. coli isolates from wild waterfowl were classified as non-multidrug-resistant (non-MDR) forms. Analysis of antibiotic sensitivity phenotypes showed that only four antibiotic-resistant phenotypes were detected among non-MDR bacteria, whereas among the MDR bacteria, two antibiotic-resistant phenotypes were detected in mallards and six in domestic waterfowl. Conclusion: The results of this study showed that wild waterfowl in Ukraine, which live in natural conditions and do not receive any antimicrobial drugs, are carriers of E. coli that are resistant to a number of antibiotics that are actively used in industrial poultry.

Complete Genome Sequence of an Avian Orthoavulavirus 13 Strain Detected in Ukraine.

We report the complete genome sequence of an avian orthoavulavirus 13 strain, isolated from a white-fronted goose in the Odesa region of Ukraine in 2013. The detection of avian orthoavulavirus 13 in Ukraine confirms that the geographic distribution of this virus extends beyond Asia.

Discovery of Avian Paramyxoviruses APMV-1 and APMV-6 in Shorebirds and Waterfowl in Southern Ukraine.

Emerging RNA virus infections are a growing concern among domestic poultry industries due to the severe impact they can have on flock health and economic livelihoods. Avian paramyxoviruses (APMV; avulaviruses, AaV) are pathogenic, negative-sense RNA viruses that cause serious infections in the respiratory and central nervous systems. APMV was detected in multiple avian species during the 2017 wild bird migration season in Ukraine and studied using PCR, virus isolation, and sequencing. Of 4090 wild bird samples collected, mostly from southern Ukraine, eleven isolates were grown in ovo and identified for APMV serotype by hemagglutinin inhibition test as: APMV-1, APMV-4, APMV-6, and APMV-7. To build One Health's capacity to characterize APMV virulence and analyze the potential risks of spillover to immunologically naïve populations, we sequenced virus genomes in veterinary research labs in Ukraine using a nanopore (MinION) platform. RNA was extracted and amplified using a multiplex tiling primer approach to specifically capture full-length APMV-1 (n = 5) and APMV-6 (n = 2) genomes at high read depth. All APMV-1 and APMV-6 fusion (F) proteins possessed a monobasic cleavage site, suggesting these APMVs were likely low virulence, annually circulating strains. Utilization of this low-cost method will identify gaps in viral evolution and circulation in this understudied but important critical region for Eurasia.

Genetic diversity of Newcastle disease viruses circulating in wild and synanthropic birds in Ukraine between 2006 and 2015.

Newcastle disease virus (NDV) infects a wide range of bird species worldwide and is of importance to the poultry industry. Although certain virus genotypes are clearly associated with wild bird species, the role of those species in the movement of viruses and the migratory routes they follow is still unclear. In this study, we performed a phylogenetic analysis of nineteen NDV sequences that were identified among 21,924 samples collected from wild and synanthropic birds from different regions of Ukraine from 2006 to 2015 and compared them with isolates from other continents. In synanthropic birds, NDV strains of genotype II, VI, VII, and XXI of class II were detected. The fusion gene sequences of these strains were similar to strains detected in birds from different geographical regions of Europe and Asia. However, it is noteworthy to mention the isolation of vaccine viruses from synanthropic birds, suggesting the possibility of their role in viral transmission from vaccinated poultry to wild birds, which may lead to the further spreading of vaccine viruses into other regions during wild bird migration. Moreover, here we present the first publicly available complete NDV F gene from a crow (genus Corvus). Additionally, our phylogenetic results indicated a possible connection of Ukrainian NDV isolates with genotype XXI strains circulating in Kazakhstan. Among strains from wild birds, NDVs of genotype 1 of class I and genotype I of class II were detected. The phylogenetic analysis highlighted the possible exchange of these NDV strains between wild waterfowl from the Azov-Black Sea region of Ukraine and waterfowl from different continents, including Europe, Asia, and Africa.

Genetic Diversity of Porcine Circovirus 2 in Wild Boar and Domestic Pigs in Ukraine.

Porcine circovirus type 2 (PCV2) is responsible for a number of porcine circovirus-associated diseases (PCVAD) that can severely impact domestic pig herds. For a non-enveloped virus with a small genome (1.7 kb ssDNA), PCV2 is remarkably diverse, with eight genotypes (a-h). New genotypes of PCV2 can spread through the migration of wild boar, which are thought to infect domestic pigs and spread further through the domestic pig trade. Despite a large swine population, the diversity of PCV2 genotypes in Ukraine has been under-sampled, with few PCV2 genome sequences reported in the past decade. To gain a deeper understanding of PCV2 genotype diversity in Ukraine, samples of blood serum were collected from wild boars (n = 107) that were hunted in Ukraine during the November-December 2012 hunting season. We found 34/107 (31.8%) prevalence of PCV2 by diagnostic PCR. For domestic pigs, liver samples (n = 16) were collected from a commercial market near Kharkiv in 2019, of which 6 out of 16 (37%) samples were positive for PCV2. We sequenced the genotyping locus ORF2, a gene encoding the PCV2 viral capsid (Cap), for 11 wild boar and six domestic pig samples in Ukraine using an Oxford Nanopore MinION device. Of 17 samples with resolved genotypes, the PCV2 genotype b was the most common in wild boar samples (10 out of 11, 91%), while the domestic pigs were infected with genotypes b and d. We also detected genotype b/d and b/a co-infections in wild boars and domestic pigs, respectively, and for the first time in Ukraine we detected genotype f in a wild boar from Poltava. Building a maximum-likelihood phylogeny, we identified a sublineage of PCV2 genotype b infections in both wild and domestic swine, suggesting a possible epizootic cluster and an ecological interaction between wild boar and domestic pig populations in northeastern Ukraine.

Zoonotic and Reverse Zoonotic Transmissibility of SARS-CoV-2.

The Coronavirus Disease 2019 (COVID-19) is the first known pandemic caused by a coronavirus. Its causative agent, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), appears to be capable of infecting different mammalian species. Recent detections of this virus in pet, zoo, wild, and farm animals have compelled inquiry regarding the zoonotic (animal-to-human) and reverse zoonotic (human-to-animal) transmissibility of SARS-CoV-2 with the potential of COVID-19 pandemic evolving into a panzootic. It is important to monitor the global spread of disease and to assess the significance of genomic changes to support prevention and control efforts during a pandemic. An understanding of the SARS-CoV-2 epidemiology provides opportunities to prevent the risk of repeated re-infection of humans and requires a robust One Health-based investigation. This review paper describes the known properties and the existing gaps in scientific knowledge about the zoonotic and reverse zoonotic transmissibility of the novel virus SARS-CoV-2 and the COVID-19 disease it causes.

Age and Seasonal Pattern of Contagious Agalactia in Small Ruminants in Ukraine.

INTRODUCTION: The aim of the study was to determine how the spread of contagious agalactia in sheep and goats in the Odesa region depended on the age of the animals and the season. MATERIAL AND METHODS: From January 2016 to December 2018, 1,964 ewes and 1,484 nanny goats of different age groups were studied by ELISA for antibodies to Mycoplasma agalactiae. RESULTS: The highest incidence of contagious agalactia was registered in one-year-old animals and was 59.7‒83.0%, two-year-old ruminants showed 17.0‒40.3% prevalence, in livestock at the age of 3-4 years no serological evidence of the disease was registered and in ewes and nanny goats older than 5-6 years 1.5-3.6% were infected. The most susceptible were young animals at the age of one-month (11.6‒14.5%). The first peak of the disease was recorded in March‒April (21.0‒26.1%), in the lambing period, which coincided with the beginning of lactation and the suckling period, and the second peak occurred in June-July (28.9‒34.2%), the period of maximum lactation and of manual milking of sheep and goats. CONCLUSION: The results of serological investigations indicate the circulation of M. agalactiae in small ruminants in the south of Ukraine. To avoid greater dissemination of the pathogen, appropriate measures should be applied and strategies for its control need to be drawn up.

Highly Pathogenic and Low Pathogenic Avian Influenza H5 Subtype Viruses in Wild Birds in Ukraine.

There have been three waves of highly pathogenic avian influenza (HPAI) outbreaks in commercial, backyard poultry, and wild birds in Ukraine. The first (2005-2006) and second (2008) waves were caused by H5N1 HPAI virus, with 45 outbreaks among commercial poultry (chickens) and backyard fowl (chickens, ducks, and geese) in four regions of Ukraine (AR Crimea, Kherson, Odesa, and Sumy Oblast). H5N1 HPAI viruses were isolated from dead wild birds: cormorants (Phalacrocorax carbo) and great crested grebes (Podiceps cristatus) in 2006 and 2008. The third HPAI wave consisted of nine outbreaks of H5N8 HPAI in wild and domestic birds, beginning in November 2016 in the central and south regions (Kherson, Odesa, Chernivtsi, Ternopil, and Mykolaiv Oblast). H5N8 HPAI virus was detected in dead mute swans (Cygnus olor), peacocks (Pavo cristatus) (in zoo), ruddy shelducks (Tadorna ferruginea), white-fronted geese (Anser albifrons), and from environmental samples in 2016 and 2017. Wide wild bird surveillance for avian influenza (AI) virus was conducted from 2006 to 2016 in Ukraine regions suspected of being intercontinental (north-south and east-west) flyways. A total of 21 511 samples were collected from 105 species of wild birds representing 27 families and 11 orders. Ninety-five avian influenza (AI) viruses were isolated (including one H5N2 LPAI virus in 2010) from wild birds with a total of 26 antigenic hemagglutinin (HA) and neuraminidase (NA) combinations. Fifteen of 16 known avian HA subtypes were isolated. Two H5N8 HPAI viruses (2016-2017) and two H5N2 LPAI viruses (2016) were isolated from wild birds and environmental samples (fresh bird feces) during surveillance before the outbreak in poultry in 2016-2017. The Ukrainian H5N1, H5N8 HPAI, and H5N2 LPAI viruses belong to different H5 phylogenetic groups. Our results demonstrate the great diversity of AI viruses in wild birds in Ukraine, as well as the importance of this region for studying the ecology of avian influenza.

Virus de influenza aviar del subtipo H5 altamente patógenos y de baja patogenicidad en aves silvestres en Ucrania. Ha habido tres oleadas de brotes de influenza aviar altamente patógena en aves comerciales, de traspatio y en aves silvestres en Ucrania. La primera (2005-2006) y la segunda (2008) fueron causadas por el virus de influenza aviar de alta patogenicidad H5N1, con 45 brotes en aves comerciales (pollos) y aves de traspatio (pollos, patos y gansos) en cuatro regiones de Ucrania (AR Crimea, Kherson, Odesa y Sumy Oblast). Los virus de alta patogenicidad H5N1se aislaron de aves silvestres muertas: cormoranes (Phalacrocorax carbo) y de somormujos lavanco (Podiceps cristatus) en 2006 y 2008. La tercera ola del virus de influenza aviar de alta patogenicidad consistió en nueve brotes del virus de alta patogenicidad subtipo H5N8 en aves silvestres y domésticas, a partir de noviembre de 2016 en las regiones central y sur (Kherson, Odesa, Chernivtsi, Ternopil y Mykolaiv Oblast). Se detectó el virus al patogenicidad H5N8 en cisnes blancos muertos (Cygnus olor), pavos reales (Pavo cristatus) (en zoológicos), tarros canelos (Tadorna ferruginea), gansos caretos (Anser albifrons) y en muestras ambientales en 2016 y 2017. Una vigilancia más amplia de aves silvestres para detectar el virus de la influenza aviar se realizó entre 2006 y 2016 en las regiones de Ucrania sospechosas de ser rutas migratorias intercontinentales (norte-sur y este-oeste). Se recolectaron un total de 21,511 muestras de 105 especies de aves silvestres que representan a 27 familias y 11 órdenes. Se aislaron ochenta y dos virus de influenza aviar de baja patogenicidad (incluido un virus H5N2 de baja patogenicidad del 2010) de aves silvestres con un total de 23 combinaciones antigénicas de hemaglutininas (HA) y neuraminidasas (NA). Se aislaron quince de los 16 subtipos de HA aviar conocidos. Dos virus de alta patogenicidad H5N8 y dos virus H5N2 de baja patogenicidad se aislaron de aves silvestres vivas y de muestras ambientales (heces de aves frescas) durante la vigilancia antes del brote en avicultura. Los virus ucranianos de alta patogenicidad H5N1, H5N8 y de baja patogenicidad H5N2 pertenecen a diferentes grupos filogenéticos de H5. Estos resultados demuestran la gran diversidad de virus de la influenza aviar en aves silvestres en Ucrania, así como la importancia de esta región para estudiar la ecología de la influenza aviar.

Highly Pathogenic and Low Pathogenic Avian Influenza H5 Subtype Viruses in Wild Birds in Ukraine.

There have been three waves of highly pathogenic avian influenza (HPAI) outbreaks in commercial, backyard poultry, and wild birds in Ukraine. The first (2005-2006) and second (2008) waves were caused by H5N1 HPAI virus, with 45 outbreaks among commercial poultry (chickens) and backyard fowl (chickens, ducks, and geese) in four regions of Ukraine (AR Crimea, Kherson, Odesa, and Sumy Oblast). H5N1 HPAI viruses were isolated from dead wild birds: cormorants (Phalacrocorax carbo) and great crested grebes (Podiceps cristatus) in 2006 and 2008. The third HPAI wave consisted of nine outbreaks of H5N8 HPAI in wild and domestic birds, beginning in November 2016 in the central and south regions (Kherson, Odesa, Chernivtsi, Ternopil, and Mykolaiv Oblast). H5N8 HPAI virus was detected in dead mute swans (Cygnus olor), peacocks (Pavo cristatus) (in zoo), ruddy shelducks (Tadorna ferruginea), white-fronted geese (Anser albifrons), and from environmental samples in 2016 and 2017. Wide wild bird surveillance for avian influenza (AI) virus was conducted from 2006 to 2016 in Ukraine regions suspected of being intercontinental (north-south and east-west) flyways. A total of 21 511 samples were collected from 105 species of wild birds representing 27 families and 11 orders. Ninety-five avian influenza (AI) viruses were isolated (including one H5N2 LPAI virus in 2010) from wild birds with a total of 26 antigenic hemagglutinin (HA) and neuraminidase (NA) combinations. Fifteen of 16 known avian HA subtypes were isolated. Two H5N8 HPAI viruses (2016-2017) and two H5N2 LPAI viruses (2016) were isolated from wild birds and environmental samples (fresh bird feces) during surveillance before the outbreak in poultry in 2016-2017. The Ukrainian H5N1, H5N8 HPAI, and H5N2 LPAI viruses belong to different H5 phylogenetic groups. Our results demonstrate the great diversity of AI viruses in wild birds in Ukraine, as well as the importance of this region for studying the ecology of avian influenza.

Virus de influenza aviar del subtipo H5 altamente patógenos y de baja patogenicidad en aves silvestres en Ucrania. Ha habido tres oleadas de brotes de influenza aviar altamente patógena en aves comerciales, de traspatio y en aves silvestres en Ucrania. La primera (2005-2006) y la segunda (2008) fueron causadas por el virus de influenza aviar de alta patogenicidad H5N1, con 45 brotes en aves comerciales (pollos) y aves de traspatio (pollos, patos y gansos) en cuatro regiones de Ucrania (AR Crimea, Kherson, Odesa y Sumy Oblast). Los virus de alta patogenicidad H5N1se aislaron de aves silvestres muertas: cormoranes (Phalacrocorax carbo) y de somormujos lavanco (Podiceps cristatus) en 2006 y 2008. La tercera ola del virus de influenza aviar de alta patogenicidad consistió en nueve brotes del virus de alta patogenicidad subtipo H5N8 en aves silvestres y domésticas, a partir de noviembre de 2016 en las regiones central y sur (Kherson, Odesa, Chernivtsi, Ternopil y Mykolaiv Oblast). Se detectó el virus al patogenicidad H5N8 en cisnes blancos muertos (Cygnus olor), pavos reales (Pavo cristatus) (en zoológicos), tarros canelos (Tadorna ferruginea), gansos caretos (Anser albifrons) y en muestras ambientales en 2016 y 2017. Una vigilancia más amplia de aves silvestres para detectar el virus de la influenza aviar se realizó entre 2006 y 2016 en las regiones de Ucrania sospechosas de ser rutas migratorias intercontinentales (norte-sur y este-oeste). Se recolectaron un total de 21,511 muestras de 105 especies de aves silvestres que representan a 27 familias y 11 órdenes. Se aislaron ochenta y dos virus de influenza aviar de baja patogenicidad (incluido un virus H5N2 de baja patogenicidad del 2010) de aves silvestres con un total de 23 combinaciones antigénicas de hemaglutininas (HA) y neuraminidasas (NA). Se aislaron quince de los 16 subtipos de HA aviar conocidos. Dos virus de alta patogenicidad H5N8 y dos virus H5N2 de baja patogenicidad se aislaron de aves silvestres vivas y de muestras ambientales (heces de aves frescas) durante la vigilancia antes del brote en avicultura. Los virus ucranianos de alta patogenicidad H5N1, H5N8 y de baja patogenicidad H5N2 pertenecen a diferentes grupos filogenéticos de H5. Estos resultados demuestran la gran diversidad de virus de la influenza aviar en aves silvestres en Ucrania, así como la importancia de esta región para estudiar la ecología de la influenza aviar.

Repeated isolation of virulent Newcastle disease viruses of sub-genotype VIId from backyard chickens in Bulgaria and Ukraine between 2002 and 2013.

Here, we report the circulation of highly related virulent Newcastle disease viruses (NDV) in Bulgaria and Ukraine from 2002 until 2013. All of these NDV isolates have the same virulence-associated cleavage site ("113RQKR↓F117"), and selected ones have intracerebral pathogenicity index values ranging from 1.61 to 1.96. These isolates are most closely related to viruses circulating in Eastern Europe, followed by viruses isolated in Asia during the same period of time. Interestingly, the majority of the viruses were isolated from backyard poultry, suggesting the possibility of a "domestic" or "urban" cycle of maintenance. The molecular characterization of the nucleotide sequence of the complete fusion protein gene of the studied viruses suggests continued circulation of virulent NDV of sub-genotype VIId in Eastern Europe, with occasional introductions from Asia. Furthermore, the high level of genetic similarity among those isolates suggests that the NDV isolates of sub-genotype VIId from Bulgaria and Ukraine may have been part of a broader epizootic process in Eastern Europe rather than separate introductions from Asia or Africa. The continuous monitoring of backyard poultry flocks for the presence of circulating virulent NDV strains will allow early identification of Newcastle disease outbreaks.

Complete Genome Sequence of an Avian Paramyxovirus Representative of Putative New Serotype 13.

Here, we report the complete genome sequence of a virus of a putative new serotype of avian paramyxovirus (APMV). The virus was isolated from a white-fronted goose in Ukraine in 2011 and designated white-fronted goose/Ukraine/Askania-Nova/48-15-02/2011. The genomic characterization of the isolate suggests that it represents the novel avian paramyxovirus group APMV 13.

Isolation and Genetic Characterization of Avian Influenza Viruses Isolated from Wild Birds in the Azov-Black Sea Region of Ukraine (2001-2012).

Wild bird surveillance for avian influenza virus (AIV) was conducted from 2001 to 2012 in the Azov - Black Sea region of the Ukraine, considered part of the transcontinental wild bird migration routes from northern Asia and Europe to the Mediterranean, Africa, and southwest Asia. A total of 6281 samples were collected from wild birds representing 27 families and eight orders for virus isolation. From these samples, 69 AIVs belonging to 15 of the 16 known hemagglutinin (HA) subtypes and seven of nine known neuraminidase (NA) subtypes were isolated. No H14, N5, or N9 subtypes were identified. In total, nine H6, eight H1, nine H5, seven H7, six H11, six H4, five H3, five H10, four H8, three H2, three H9, one H12, one H13, one H15, and one H16 HA subtypes were isolated. As for the NA subtypes, twelve N2, nine N6, eight N8, seven N7, six N3, four N4, and one undetermined were isolated. There were 27 HA and NA antigen combinations. All isolates were low pathogenic AIV except for eight highly pathogenic (HP) AIVs that were isolated during the H5N1 HPAI outbreaks of 2006-08. Sequencing and phylogenetic analysis of the HA genes revealed epidemiological connections between the Azov-Black Sea regions and Europe, Russia, Mongolia, and Southeast Asia. H1, H2, H3, H7, H8, H6, H9, and H13 AIV subtypes were closely related to European, Russian, Mongolian, and Georgian AIV isolates. H10, H11, and H12 AIV subtypes were epidemiologically linked to viruses from Europe and Southeast Asia. Serology conducted on serum and egg yolk samples also demonstrated previous exposure of many wild bird species to different AIVs. Our results demonstrate the great genetic diversity of AIVs in wild birds in the Azov-Black Sea region as well as the importance of this region for monitoring and studying the ecology of influenza viruses. This information furthers our understanding of the ecology of avian influenza viruses in wild bird species.

Wild bird surveillance for avian paramyxoviruses in the Azov-black sea region of Ukraine (2006 to 2011) reveals epidemiological connections with Europe and Africa.

Despite the existence of 10 avian paramyxovirus (APMV) serotypes, very little is known about the distribution, host species, and ecological factors affecting virus transmission. To better understand the relationship among these factors, we conducted APMV wild bird surveillance in regions of Ukraine suspected of being intercontinental (north to south and east to west) flyways. Surveillance for APMV was conducted in 6,735 wild birds representing 86 species and 8 different orders during 2006 to 2011 through different seasons. Twenty viruses were isolated and subsequently identified as APMV-1 (n = 9), APMV-4 (n = 4), APMV-6 (n = 3), and APMV-7 (n = 4). The highest isolation rate occurred during the autumn migration (0.61%), with viruses isolated from mallards, teals, dunlins, and a wigeon. The rate of isolation was lower during winter (December to March) (0.32%), with viruses isolated from ruddy shelducks, mallards, white-fronted geese, and a starling. During spring migration, nesting, and postnesting (April to August) no APMV strains were isolated out of 1,984 samples tested. Sequencing and phylogenetic analysis of four APMV-1 and two APMV-4 viruses showed that one APMV-1 virus belonging to class 1 was epidemiologically linked to viruses from China, three class II APMV-1 viruses were epidemiologically connected with viruses from Nigeria and Luxembourg, and one APMV-4 virus was related to goose viruses from Egypt. In summary, we have identified the wild bird species most likely to be infected with APMV, and our data support possible intercontinental transmission of APMVs by wild birds.

Avian influenza virus wild bird surveillance in the Azov and Black Sea regions of Ukraine (2010-2011).

The Azov and Black Sea basins are part of the transcontinental wild bird migration routes from Northern Asia and Europe to the Mediterranean, Africa, and Southwest Asia. These regions constitute an area of transit, stops during migration, and nesting for many different bird species. From September 2010 to September 2011, a wild bird surveillance study was conducted in these regions to identify avian influenza viruses. Biological samples consisting of cloacal and tracheal swabs and fecal samples were collected from wild birds of different ecological groups, including waterfowl and sea- and land-based birds, in places of mass bird accumulations in Sivash Bay and the Utlyuksky and Molochniy estuaries. The sampling covered the following wild bird biological cycles: autumn migration, wintering, spring migration, nesting, and postnesting seasons. A total of 3634 samples were collected from 66 different species of birds. During the autumn migration, 19 hemagglutinating viruses were isolated, 14 of which were identified as low pathogenicity avian influenza (LPAI) virus subtypes H1N?, H3N8, H5N2, H7N?, H8N4, H10N7, and H11N8. From the wintering samples, 45 hemagglutinating viruses were isolated, 36 of which were identified as LPAI virus subtypes H1N1, H1N? H1N2, H4N?, H6N1, H7N3, H7N6, H7N7, H8N2, H9N2, H10N7, H10N4, H11N2, H12N2, and H15N7. Only three viruses were isolated during the spring migration, nesting, and postnesting seasons (serotypes H6, H13, and H16). The HA and NA genes were sequenced from the isolated H5 and N1 viruses, and the phylogenetic analysis revealed possible ecological connections between the Azov and Black Sea regions and Europe. The LPAI viruses were isolated mostly from mallard ducks, but also from shellducks, shovelers, teals, and white-fronted geese. The rest of the 14 hemagglutinating viruses isolated were identified as different serotypes of avian paramyxoviruses (APMV-1, APMV-4, APMV-6, and APMV-7). This information furthers our understanding of the ecology of avian influenza viruses in wild bird species.

The bovine tuberculosis burden in cattle herds in zones with low dose radiation pollution in Ukraine.

The authors describe a study of the tuberculosis (TB) incidence in cattle exposed to low doses of radiation resulting from the Chernobyl (pronounced 'Chornobyl' in Ukrainian) nuclear plant catastrophe in 1986. The purpose of the study was to determine if ionising radiation influences the number of outbreaks of bovine TB and their severity on farms in the Kyiv, Cherkasy and Chernigiv regions of Ukraine. These farms are all located within a 200 km radius of Chernobyl and have had low-dose radiation pollution. Pathological and blood samples were taken from cattle in those regions that had positive TB skin tests. Mycobacterium spp. were isolated, differentiated by PCR, analysed and tested in guinea-pigs and rabbits. Species differentiation showed a significant percentage of atypical mycobacteria, which resulted in the allergic reactions to tuberculin antigen in the skin test. Mixed infection of M. bovis and M. avium subsp. hominissuis was found in three cases. The results concluded that low-dose radiation plays a major role in the occurrence of bovine TB in regions affected by the Chernobyl nuclear disaster.