Detection of a range of genetically diverse chlamydiae in Australian domesticated and wild ungulates

Chlamydiae are globally widespread obligate intracellular bacteria, which several species are a well‐recognized threat to human and animal health. In Australia, the most successful chlamydial species are the infamous koala pathogen C. pecorum, and C. psittaci, an emerging pathogen associated with zoonotic events. Little is known about infections caused by other chlamydial organisms in Australian livestock or wildlife. Considering that these hosts can be encountered by humans at the animal/human interface, in this study, we investigated genetic diversity of chlamydial organisms infecting Australian domesticated and wild ungulates. A total of 185 samples from 129 domesticated (cattle, horses, sheep, and pigs) and 29 wild (deer) ungulate hosts were screened with C. pecorum and C. psittaci species‐specific assays, followed by a screen with pan‐Chlamydiales assay. Overall, chlamydial DNA was detected in 120/185 (65%) samples, including all ungulate hosts. Species‐specific assays further revealed that C. pecorum and C. psittaci DNA were detected in 27% (50/185) and 6% (11/185) of the samples, respectively, however from domesticated hosts only. A total of 46 “signature” 16S rRNA sequences were successfully resolved by sequencing and were used for phylogenetic analyses. Sequence analyses revealed that genetically diverse novel as well as traditional chlamydial organisms infect an expanded range of ungulate hosts in Australia. Detection of the C. psittaci and C. pecorum in livestock, and novel taxa infecting horses and deer raises questions about the genetic make‐up and pathogenic potential of these organisms, but also concerns about risks of spill‐over between livestock, humans, and native wildlife.     

Molecular epidemiological characterization of Brucella isolates from sheep and yaks in northwest China

Animal brucellosis is a re‐emerging disease in China with high prevalence in the northwest region. A total of 66 isolates of Brucella were recovered from sheep and yaks in the Inner Mongolia, Xinjiang, Qinghai and Gansu provinces of northwest China in 2015 and 2016. Using classical biotyping and the Brucella AMOS PCR assay, all isolates were identified as Brucella melitensis biovar 3 (n  = 58), B. melitensis biovar 1 (n  = 1), Brucella abortus (n  = 5), or Brucella suis biovar 3 (n  = 2), and B. melitensis biovar 3 was found to be mainly responsible for sheep brucellosis in northwest China. Multilocus variable‐number tandem‐repeat analysis (MLVA ) was used to identify the epidemiological relationships among the isolates and to assess their genetic diversity. Multilocus variable‐number tandem‐repeat analysis‐16 identified 46 genotypes in these populations, including 37 unique and nine shared genotypes. Multilocus variable‐number tandem‐repeat analysis‐11 showed that 71% of the isolates (47 of 66) were genotype 116 (1‐5‐3‐13‐2‐2‐3‐2‐4‐41‐8), a characteristic subgroup of the East Mediterranean group, showing that isolates from different geographical areas exhibit similar epidemiological characteristics in different regions and may be epidemiologically linked. Multilocus variable‐number tandem‐repeat analysis‐11 also revealed that an isolate from Inner Mongolia had a novel genotype, 369 (1‐5‐3‐13‐2‐2‐3‐2‐7‐41‐8). Multilocus variable‐number tandem‐repeat analysis‐16 genotyping of northwest China Brucella isolates allows a better understanding of the epidemiology of animal brucellosis in this region. This study is the first analysis of B. melitensis in Gansu province, and the results confirmed that in this province, isolates of this species are disorderly and unsystematic.     

A wild circulation: High presence of Porcine circovirus 3 in different mammalian wild hosts and ticks

Porcine circovirus 3 (PCV‐3) has emerged as a potential threat for swine industry, being consistently reported in the presence of several clinical signs all around the world. Recently, its presence in wild boar has been demonstrated at high prevalence. This evidence is surprising since the lower density of wild populations might not be expected to sustain such efficient viral transmission. Porcine circoviruses were proven to exhibit a certain plasticity in the host tropism and were detected in unrelated species, like mice, dogs and ruminants. However, if this scenario applies also to wild animals remains to be established. Therefore, this study aimed to investigate the presence of PCV‐3 in wild ungulates other than wild boar and in related hematophagous ectoparasites. One hundred and nine animals were sampled from different hilly and mountain areas of Friuli Venezia Giulia, including 9 chamois (Rupicapra rupicapra), 17 red deer (Cervus elaphus), 4 mouflons (Ovis musimon), 50 roe deer (Capreolus capreolus) and 29 wild boars (Sus scrofa). Additionally, host‐matched ectoparasites were collected when present. Porcine circovirus 3 was diagnosed using molecular techniques and sequencing. This study results confirmed the high PCV‐3 occurrence in wild boar and reported for the first time its presence, at low prevalence, in chamois and roe deer. Moreover, two ticks (Ixodes ricinus), one of which non‐engorged, collected from PCV‐3 negative roe deer, tested PCV‐3 positive. The genetic characterization of some of the strains collected from non‐swine hosts allowed to prove that, albeit clearly part of PCV‐3 species, they were genetically unique, demonstrating the absence of among‐samples contamination and thus confirming the actual presence of PCV‐3 genome in these new hosts. Therefore, this study highlights an unexpected broad PCV‐3 distribution and circulation in the wild, rising further questions on porcine circoviruses infectious cycle, epidemiology and origin, which will deserve additional investigations.     

Free‐living Waterfowl as a Source of Zoonotic Bacteria in a Dense Wild Bird Population Area in Northeastern Spain

Salmonella spp. and Campylobacter spp. are zoonotic bacteria that represent an economic and public health concern worldwide. Due to the difficulty to collect samples from free‐living waterfowl, little is known on their importance as a reservoir of zoonotic agents. Thus, a study was conducted to determine the prevalence, genotypic diversity and antimicrobial susceptibility of Salmonella and Campylobacter from waterfowl in Ebro Delta (northeastern Spain), a geographical area with a dense wild bird population. Samples were collected from 318 adult waterfowl belonging to nine fowl species. All the samples were taken during the hunting season from 2008 to 2010. None of the birds were positive for Salmonella, while the overall Campylobacter prevalence was 12.58% (40/318). A much higher Campylobacter coli prevalence than Campylobacter jejuni was found (11.64% versus 0.94%). The species Fulica atra showed the highest Campylobacter prevalence (78.05%). ERIC‐PCR of the isolates showed a high diversity of strains. Antimicrobial susceptibility testing of Campylobacter isolates showed that all the isolates were susceptible to the seven antibiotics tested.     

Rickettsia parkeri in free‐ranging wild canids from Brazilian Pampa

Spotted fevers are tick‐borne diseases associated with various Rickettsia species. Rickettsia parkeri sensu stricto (s.s.) is the agent of an emerging eschar‐associated rickettsiosis in humans from the USA and South American Pampa. Considering that R. parkeri s.s. is restricted to Americas and the potential role of dogs in the epidemiology of the disease, it is thus reasonable to hypothesize that wild canids could be involved in the enzootic cycle of this rickettsiosis. The aim of this work was to investigate the potential role of the wild canids from Pampa, Cerdocyon thous (crab‐eating fox) and Lycalopex gymnocercus (Pampas fox), in the ecology of R. parkeri s.s. For that, 32 live‐trapped free‐ranging wild canids were sampled. Ticks were observed in 30 of the 32 foxes. Of the 292 ticks collected, 22 (7.5%) were positive by PCR for the presence of R. parkeri s.s. DNA . Also, 20 (62%) wild canids showed antibodies against R. parkeri . The results suggest that wild canids are involved in the enzootic cycle of R. parkeri s.s. in the Pampa biome and could be responsible for pathogen (and its vectors) dispersal.     

Genetic characterization of African swine fever virus isolates from soft ticks at the wildlife/domestic interface in Mozambique and identification of a novel genotype

African swine fever virus (ASFV ) is one of the most threatening infectious diseases of pigs. There are not sufficient data to indicate the importance of the sylvatic cycle in the spread and maintenance of the disease locally and potentially, globally. To assess the capacity to maintain ASF in the environment, we investigated the presence of soft tickreservoirs of ASFV in Gorongosa National Park (GNP ) and its surrounding villages. A total of 1,658 soft ticks were recovered from warthog burrows and pig pens at the wildlife/livestock interface of the GNP and viral DNA was confirmed by nested PCR in 19% of Ornithodoros porcinus porcinus and 15% of O. p. domesticus . However, isolation of ASFV was only achieved in approximately 50% of the PCR ‐positive samples with nineteen haemadsorbing virus isolates recovered. These were genotyped using a combination of partial sequencing of the B646L gene (p72 ) and analysis of the central variable region (CVR ) of the B602L gene. Eleven isolates were classified as belonging to genotype II and homologous to contemporary isolates from southern Africa, the Indian Ocean and eastern Europe. Three isolates grouped within genotype V and were similar to previous isolates from Mozambique and Malawi. The remaining five isolates constituted a new, previously unidentified genotype, designated genotype XXIV . This work confirms for the first time that the virus currently circulating in eastern Europe is likely to have a wildlife origin, and that the large diversity of ASFV maintained in wildlife areas can act as a permanent sources of different strains for the domestic pig value chain in Mozambique and beyond its boundaries. Their genetic similarity to ASFV strains currently spreading across Europe justifies the need to continue studying the sylvatic cycle in this African country and other parts of southern Africa in order to identify potential hot spots of ASF emergence and target surveillance and control efforts.     

Coxiella burnetii in wild mammals: A systematic review

Coxiella burnetii is a multi‐host bacterium that causes Q fever in humans, a zoonosis that is emerging worldwide. The ecology of C. burnetii in wildlife is still poorly understood and the influence of host, environmental and pathogen factors is almost unknown. This study gathers current published information on different aspects of C. burnetii infection in wildlife, even in species with high reservoir potential and a high rate of interaction with livestock and humans, in order to partially fill the existing gap and highlight future needs. Exposure and/or infection by C. burnetii has, to date, been reported in 109 wild mammal species. The limited sample size of most of the existing studies could suggest an undervalued prevalence of C. burnetii infection. Knowledge on the clinical outcome of C. burnetii infection in wildlife is also very limited, but currently includes reproductive failure in waterbuck (Kobus ellipsiprymnus), roan antelope (Hippotragus niger), dama gazelle (Nanger dama) and water buffalo (Bubalus bubalis) and placentitis in the Pacific harbor seal (Phoca vitulina richardsi), Steller sea lion (Eumetopias jubatus) and red deer (Cervus elaphus). The currently available serological tests need to be optimised and validated for each wildlife species. Finally, there is a huge gap in the research on C. burnetii control in wildlife, despite of the increasing evidence that wildlife is a source of C. burnetii for both livestock and humans.     

Genetic diversity of Bartonella spp. in vampire bats from Brazil

Recently, an increasing number of Bartonella species have been emerged to cause human diseases. Among animal reservoirs for Bartonella spp., bats stand out due to their high mobility, wide distribution, social behaviour and long‐life span. Although studies on the role of vampire bats in the epidemiology of rabies have been extensively investigated in Latin America, information on the circulation and genetic diversity of Bartonella species in these bat species is scarce. In the present work, 208 vampire bats, namely Desmodus rotundus (the common vampire bat; n = 167), Diphylla ecaudata (the hairy‐legged vampire bat; n = 32) and Diaemus youngii (the white‐winged vampire bat; n = 9) from 15 different states in Brazil were sampled. DNA was extracted from liver tissue samples and submitted to real‐time PCR (qPCR) and conventional PCR (cPCR) assays for Bartonella spp. targeting five genetic loci, followed by phylogenetic and genotype network analyses. Fifty‐one out of 208 liver samples (24.51%) were positive for Bartonella DNA in the ITS real‐time PCR assay [40 (78.43%) of them were from D. rotundus from 11 states, and 11 (21.57%) samples from D. ecaudata from three states. Eleven genotypes were found for each gltA and rpoB genes. Several ITS sequences detected in the present study clustered within the lineage that includes B. bacilliformis and B. ancachensis. The Bayesian phylogenetic inference based on the gltA gene positioned the obtained sequences in six different clades, closely related to Bartonella genotypes previously detected in D. rotundus and associated ectoparasites sampled in Latin America. On the other hand, the Bartonella rpoB genotypes clustered together with the ruminant species, B. schoenbuchensis and B. chomelii. The present study describes for the first time the molecular detection of Bartonella spp. in D. ecaudata bats. It also indicates that Bartonella spp. of vampire bats are genetically diverse and geographically widespread in Brazil.     

Tuberculosis in Southern Brazilian wild boars (Sus scrofa): First epidemiological findings

Bovine tuberculosis (bTB ) is a zoonosis caused mainly by Mycobacterium bovis that affects domestic and wild animals. In Brazil, there are no epidemiological studies on tuberculosis in wild animal populations and their possible role in the disease maintenance in cattle herds; thus, the aim of this study was to evaluate the occurrence of tuberculosis in wild boars in Rio Grande do Sul, southern Brazil. Tissue samples of animals hunted under government consent were submitted to histopathology and M. bovis polymerase chain reaction (PCR ) as screening tests; the positive samples were subsequently submitted to bacterial isolation, the gold standard diagnosis. Eighty animals were evaluated, of which 27.9% and 31.3% showed histopathological changes and M. bovis genome presence, respectively. Moreover, 23.8% of the animals had at least one organ with isolates classified as Mycobacterium tuberculosis complex (MTC ). Three hunting points were risk factors for positive results on screening tests. This study shows the occurrence of tuberculosis in a wild boars’ population, and raise the possibility of these animals to play a role as disease reservoirs in southern Brazil. These results may help to improve the Brazilian tuberculosis control programme, as well as elucidate the circulation of mycobacteria in this country.     

First report on characterization and pathogenicity study of emerging Lactococcus garvieae infection in farmed rainbow trout,Oncorhynchus mykiss (Walbaum), from India

“ Warm water lactococcosis” in farm‐reared rainbow trout, Oncorhynchus mykiss (Walbaum) in the northern Himalayan region of India, caused by bacterium Lactococcus garvieae is described in this study. Nine bacterial isolates were recovered from the organs of haemorrhagic septicaemia rainbow trout and were subjected to biochemical and molecular identification. Cell surface characteristics and virulence of the bacterial isolates are also described. All the nine bacterial isolates had homogenous biochemical characteristics and were Gram‐positive, short chains forming (two to eight cells long), α‐haemolytic, non‐motile ovoid cocci. Partial 16S rDNA nucleotide sequence (~1,400 bp) of current isolates shared 99% identities with the 16S rDNA nucleotide sequence of L. garvieae R421, L. garvieae FMA 395 and L. garvieae CAU :1730. The identity of the bacterial isolates was further confirmed by PCR amplification of L . garvieae‐ specific ~1,100 bp fragment. Transmission electron microscopy (TEM ) of one representative isolate, L. garvieae RTCLI 04, indicates that the isolated strain lacks thick outer capsule and is of KG + (non‐capsulates) phenotype. An intraperitoneal and intramuscular injection (2.6 × 10⁵ CFU ml⁻¹) and also immersion in bacterial suspension @ of 2.6 × 10⁵ CFU ml⁻¹ to healthy rainbow trout juveniles (body weight: 27.5 ± 3.7 g) with L . garvieae RTCLI 04 caused 80%, 60% and 10% cumulative mortality in challenged fish, respectively, within 15 days post‐infection. The haemorrhagic septicaemic disease was reproduced experimentally. Histopathological examination of organs of experimentally infected fish revealed extensive degenerative and inflammatory changes in eye, kidney, gill and liver. PCR amplification of several putative virulence genes such as haemolysins, adhesins, LP xTG ‐containing surface proteins and adhesins cluster confirms the virulence of our Indian L. garvieae isolates. To the best of our knowledge, we are reporting for the first time that L. garvieae is associated with fatal haemorrhagic septicaemia in farmed rainbow trout in India.     

Molecular detection and genetic diversity of Bartonella species in large ruminants and associated ectoparasites from the Brazilian Cerrado

Currently, five Bartonella species and an expanding number of Candidatus Bartonella species have globally been reported in ruminants. Likewise, different Bartonella genotypes were identified. However, studies relating to ruminant‐associated Bartonella in Brazil are scarce. The current study aimed to assess the prevalence and genetic diversity of Bartonella in cattle, buffaloes and associated ectoparasites in Brazil. For this purpose, EDTA‐blood samples from 75 cattle and 101 buffaloes were sampled. Additionally, 128 Rhipicephalus microplus and one Amblyomma sculptum ticks collected from cattle, and 197 R. microplus, one A. sculptum and 170 lice (Haematopinus tuberculatus) collected from buffaloes were included. Bartonella DNA was initially screened through an HRM real‐time PCR assay targeting the 16S–23S internal transcribed spacer (ITS), and the positive samples were submitted to an additional HRM assay targeting the ssrA gene. The HRM‐positive amplicons were sequenced, and the nucleotide identity was assessed by BLASTn. Bartonella spp.‐positive DNA samples were analysed by conventional PCR assays targeting the gltA and rpoB genes, and then, the samples were cloned. Finally, the phylogenetic positioning and the genetic diversity of clones were assessed. Overall, 21 of 75 (28%) cattle blood samples and 13 of 126 (10.3%) associated ticks were positive for Bartonella bovis. Out of 101 buffaloes, 95 lice and 188 tick DNA samples, one (1%) buffalo and four (4.2%) lice were positive for Bartonella spp. Conversely, none of the ticks obtained from buffaloes were positive for Bartonella. The Bartonella sequences from buffaloes showed identity ranging from 100% (ITS and gltA) to 94% (ssrA) with B. bovis. In contrast, the Bartonella DNA sequences from lice were identical (100%) to uncultured Bartonella sp. detected in cattle tail louse (Haematopinus quadripertusus) from Israel in all amplified genes. The present study demonstrates the prevalence of new B. bovis genotypes and a cattle lice‐associated Bartonella species in large ruminants and their ectoparasites from Brazil. These findings shed light on the distribution and genetic diversity of ruminant‐ and ectoparasite‐related Bartonella in Brazil.     

Genetic Characterization of Circulating African Swine Fever Viruses in Nigeria (2007–2015)

Sequencing and analysis of three discrete genome regions of African swine fever viruses (ASFV) from archival samples collected in 2007–2011 and active and passive surveillance between 2012 and 2015 in Nigeria were carried out. Analysis was conducted by genotyping of three single‐copy African swine fever (ASF) genes. The E183L and B646L genes that encode structural proteins p54 and p72, respectively, were utilized to delineate genotypes before intragenotypic resolution by characterization of the tetrameric amino acid repeat region within the hypervariable central variable region of the B602L gene. The results showed no variation in the p72 and p54 gene regions sequenced. Phylogeny of p72 sequences revealed that all the Nigerian isolates belonged to genotype I, while that of the p54 recovered the Ia genotype. Analysis of B602L gene revealed the differences in the number of tetrameric repeats. Four new variants (Tet‐15, Tet‐17a, Tet‐17b and Tet‐48) were recovered, while a fifth variant (Tet‐20) was the most widely distributed in the country displacing Tet‐36 reported previously in 2003–2006. The viruses responsible for ASF outbreaks in Nigeria are from very closely related but mutated variants of the virus that have been circulating since 1997. A practical implication of the genetic variability of the Nigerian viral isolates in this study is the need for continuous sampling and analysis of circulating viruses, which will provide epidemiological information on the evolution of ASFV in the field versus new incursion for informed strategic control of the disease in the country.     

Molecular detection and characterization of carnivore parvoviruses in free‐ranging Eurasian otters (Lutra lutra) in southern Italy

The most important Italian population of the Eurasian otter (Lutra lutra) occurs in the southern part of the peninsula with two isolated sub‐populations of about 250 adult individuals. The Eurasian otter is considered to be near threatened and it is a fully protected species. The aims of this study were to investigate for the first time the occurrence and characterize the parvoviruses included in the species Carnivore protoparvovirus 1 in seven carcasses of road‐killed Eurasian otters from the southern Italy. Carnivore protoparvovirus 1 are responsible for acute gastroenteritis and leukopenia in pets and free‐ranging carnivores. Initial screening of tissue samples by real‐time PCR revealed CPV/FPV DNA in tissue samples of five Eurasian otters; three of them, showed co‐infections by both CPV and FPV. Among the five positive Eurasian otters, we successfully obtained six DNA sequences from four individuals including two CPV‐2a, one CPV‐2b, one CPV‐2c, and two FPV sequences. Comparison of these sequences with 250 VP2 gene sequences deposited in the GenBank database, showed 10 nt differences resulting in two synonymous and eight non‐synonymous substitutions. On the basis of these results, two sequences here found were characterized as new CPV‐2a, one was characterized as new CPV‐2b variant, and one was characterized as FPV‐like mutant. The last two sequences belong to a FPV and CPV‐2c strain respectively. Carnivore protoparvovirus 1 is reported for the first time in the Eurasian otter showing high infection value in southern Italy. Occurrence of this infection should be studied further to understand its possible pathogenicity and virulence to the fragile and isolate Eurasian otter population which live in southern Italy.     

Biological characterization of African swine fever virus genotype II strains from north‐eastern Estonia in European wild boar

Due to its impact on animal health and pig industry, African swine fever (ASF) is regarded as one of the most important viral diseases of pigs. Following the ongoing epidemic in the Transcaucasian countries and the Russian Federation, African swine fever virus was introduced into the Estonian wild boar population in 2014. Epidemiological investigations suggested two different introductions into the southern and the north‐eastern part of Estonia. Interestingly, outbreak characteristics varied considerably between the affected regions. While high mortality and mainly virus‐positive animals were observed in the southern region, mortality was low in the north‐eastern area. In the latter, clinically healthy, antibody‐positive animals were found in the hunting bag and detection of virus was rare. Two hypotheses could explain the different behaviour in the north‐east: (i) the frequency of antibody detections combined with the low mortality is the tail of an older, so far undetected epidemic wave coming from the east, or (ii) the virus in this region is attenuated and leads to a less severe clinical outcome. To explore the possibility of virus attenuation, a re‐isolated ASFV strain from the north‐eastern Ida‐Viru region was biologically characterized in European wild boar. Oronasal inoculation led to an acute and severe disease course in all animals with typical pathomorphological lesions. However, one animal recovered completely and was subsequently commingled with three sentinels of the same age class to assess disease transmission. By the end of the trial at 96 days post‐initial inoculation, all animals were completely healthy and neither virus nor viral genomes were detected in the sentinels or the survivor. The survivor, however, showed high antibody levels. In conclusion, the ASFV strain from north‐eastern Estonia was still highly virulent but nevertheless, one animal recovered completely. Under the experimental conditions, no transmission occurred from the survivor to susceptible sentinel pigs.     

The Distribution of Escherichia coli Serovars,Virulence Genes,Gene Association and Combinations and Virulence Genes Encoding Serotypes in Pathogenic E. coli Recovered from Diarrhoeic Calves,Sheep and Goat

Ruminants, especially cattle, have been implicated as a principal reservoir of one of the enterovirulent Escherichia coli pathotypes. The detection of the virulence genes in diarrhoeic calves and small ruminants has not been studied in Egypt. To determine the occurrence, serotypes and the virulence gene markers, stx1, stx2, hylA, Flic_h7, stb, F41, K99, sta, F17, LT‐I, LT‐II and eae, rectal swabs were taken from diarrhoeic calves, sheep and goats and subjected to bacterial culture and PCR. The E. coli prevalence rate in the diarrhoeic animals was 63.6% in calves, 27.3% in goat and 9.1% in sheep. The 102 E. coli strains isolated from the calves, goat and sheep were 100% haemolytic non‐verotoxic and fitted into the Eagg group. The isolates belonged to seven O serogroups (O25, O78, O86, O119, O158, O164 and O157). The eae gene was detected in six of the strains isolated from the calves. The 102 bovine, ovine and caprine E. coli strains isolated in this study were negative for stx1, stx2, F41, LT‐I and Flic_h7 genes. The highest gene combinations were found to occur in the form of 24/102 isolates (23.5%) that carried the F17 gene predominantly associated with eaeA, hylA, K99 and Stb genes in the calves, while the hylA, K99 and Sta were the only genes found to be in conjunction in both calves and goats (6/102; 5.9% each). Our data show that in Egypt, large and small ruminants could be a potential source of infection in humans.     

Genetic characterization of one duck‐origin paramyxovirus type 4 strain in China

Avian paramyxovirus type 4 (APMV‐4) has been frequently reported from wildfowl and waterfowl in recent year. However, few studies have reported on the molecular characteristics and regional transmission of APMV‐4, knowledge of which is important for understanding the genetic diversity and epidemiology of avian paramyxovirus. Herein, we report the isolation of one APMV‐4 strain, designated as QY17, from the duck in eastern China. The determined complete genome of the isolate with six gene segments 3′‐N‐P‐M‐F‐HN‐L‐5′ was 15,054 nt in length. Genetic analysis of the whole‐fusion gene of this isolate showed that QY17 was derived from a Eurasian lineage. Further phylogenetic analysis showed that the duck‐origin strain QY17 had a highly genetic relationship with representative APMV‐4 strains from wildfowl in neighbouring regions. These genetic results suggested that APMV‐4 viral exchange may occur in wildfowl and poultry via wild bird migration.     

Prevalence,Isolation and Molecular Characterization of Bartonella Species in Republic of Korea

To determine the prevalence of Bartonella species and identify which species of Bartonella naturally infects the striped field mouse (Apodemus agrarius) in the Republic of Korea (ROK), spleens from 200 mice were assayed by nested polymerase chain reaction (nPCR) targeting the RNA polymerase subunit beta (rpoB) gene and the 16S‐23S internal transcribed spacer (ITS) region for members of the genus Bartonella. Utilizing PCR techniques, the prevalence of Bartonella spp. ranged from 31.5% (63/200) to 62.0% (124/200) for the rpoB and ITS gene fragments, respectively. The most prevalent species, Bartonella grahamii, was assigned to 17 genotypes and closely related to the zoonotic pathogens, B. taylorii, B. tribocorum, B. phoceensis and B. henselae, which also were detected. Two Bartonella isolates (KRBG28 and KRBG32) were recovered from blood of A. agrarius captured in Gyeonggi Province, ROK. Comparison of the 16S rRNA, hemin‐binding protein E (hbpE), glutamate dehydrogenase 1 (gdh1), invasion‐associated protein B (ialB), cell division protein (ftsZ), citrate synthase (gltA), 60 kDa heat shock protein (groEL), rpoB gene fragments and the ITS region sequences from the isolates with GenBank was confirmed as B. grahamii. Phylogenetic analysis based on the alignment of concatenated sequences (4933 bp) of KRBG28 and KRBG32 clustered with B. grahamii, forming an independent clade between Asian and American/European B. grahamii genogroups.     

Genetic characterization of canine parvovirus type 2c from domestic dogs in Korea

Canine parvovirus type 2 (CPV‐2) is an aetiological agent that causes acute haemorrhagic enteritis and fatal myocarditis in dogs. Since CPV‐2 first emerged in the late 1970s, its rapid evolution has resulted in three antigenic variants: CPV‐2a, CPV‐2b and CPV‐2c. Here, we report, for the first time in Korea, two cases of CPV‐2c infection in two dogs with severe diarrhoea. The complete open reading frame (4,269nt) of CPV‐2, encoding both non‐structural (NS) and structural (VP) proteins, was sequenced. Based on the amino acid Gln present at residue 426 of the VP2 gene, these strains were typed as CPV‐2c, and were named Korea CPV‐2c_1 and Korea CPV‐2c_2. These strains shared 99.48% reciprocal nucleotide sequence identity and had the highest nucleotide identity (99.77%–99.34%) with Asian CPV strains isolated in China, Italy (found in a dog imported from Thailand), and Vietnam from 2013 to 2017. Phylogenetic analysis based on the non‐structural (NS1) and capsid (VP2) genes revealed that Korean CPV‐2c strains clustered closely to Asian CPV strains, and separately from strains isolated in Europe, South America and North America. Amino acid changes never reported before were observed in NS1 (Thr70Pro, Cys287Tyr), VP1 (Lys17Arg, Phe33Leu) and VP2 (Gln365His, Ala516Val). Additional observed mutations, including Phe267Tyr, Tyr324Ile and Gln370Arg, have been previously reported in the recent CPV‐2c strains with Asian origins. These results suggest that the Korean CPV‐2c strains were potentially introduced via neighbouring Asian countries.