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Simple Summary: The H9N2 subtype of avian influenza virus (AIV) was first identified in turkey flocks in Wisconsin, USA, in 1966. Since then, H9N2 AIVs have been detected in mammals, domestic poultry, and wild birds. In June 2020, a new H9N2/Y280 was discovered in Korea. Existing commercial vaccines might not be fully effective against H9N2/Y280. Since influenza viruses undergo mutations rapidly, fast and accurate vaccine production is critical to address variant strains. Consequently, this study focused on rapid and effective vaccine production techniques against H9N2/Y280. The recombinant rHVT/Y280 was successfully constructed using NHEJ-CRISPR/Cas9 gene-editing technology. rHVT/Y280 vaccination induced strong humoral immunity, providing significant protection against H9N2/Y280 strain A21-MRA-003 following a challenge of 2 × 107 50% egg infectious dose. Therefore, rHVT/Y280 is a potential candidate for a live vaccine against H9N2/Y280. Since the outbreak of the H9N2/Y439 avian influenza virus in 1996, the Korean poultry industry has incurred severe economic losses. A novel possibly zoonotic H9N2 virus from the Y280-like lineage (H9N2/Y280) has been prevalent in Korea since June 2020, posing a threat to the poultry sector. Rapid mutation of influenza viruses urges the development of effective vaccines against newly generated strains. Thus, we engineered a recombinant virus rHVT/Y280 to combat H9N2/Y280. We integrated the hemagglutinin (HA) gene of the H9N2/Y280 strain into the US2 region of the herpesvirus of turkeys (HVT) Fc126 vaccine strain, utilizing CRISPR/Cas9 gene-editing technology. The successful construction of rHVT/Y280 was confirmed by polymerase chain reaction and sequencing, followed by efficacy evaluation. Four-day-old specific pathogen-free chickens received the rHVT/Y280 vaccine and were challenged with the H9N2/Y280 strain A21-MRA-003 at 3 weeks post-vaccination. In 5 days, there were no gross lesions among the vaccinated chickens. The rHVT/Y280 vaccine induced strong humoral immunity and markedly reduced virus shedding, achieving 100% inhibition of virus recovery in the cecal tonsil and significantly lowering tissue viral load. Thus, HVT vector vaccines expressing HA can be used for protecting poultry against H9N2/Y280. The induction of humoral immunity by live vaccines is vital in such cases. In summary, the recombinant virus rHVT/Y280 is a promising vaccine candidate for the protection of chickens against the H9N2/Y280. [ABSTRACT FROM AUTHOR]

Infectious bursal disease (IBD), caused by IBD virus (IBDV), is an extremely contagious immunosuppressive disease that causes major losses for the poultry industry worldwide. Recently, the novel variant IBDV (G2d) has been highly prevalent in Korea, but the current vaccines against this very virulent IBDV have limited efficacy against this novel variant. To develop a vaccine against this variant IBDV, a recombinant virus designated rHVT-VP2 was constructed by inserting the IBDV (G2d) VP2 gene into herpesvirus of turkeys (HVT) using CRISPR/Cas9 gene-editing technology. The PCR and sequencing results obtained showed that the recombinant virus rHVT-VP2 was successfully constructed. Vaccination with rHVT-VP2 generated IBDV-specific antibodies in specific pathogen-free chickens starting from 2 weeks post-immunization. Seven days after the challenge, the autopsy results showed that the bursa atrophy rates of the rHVT-VP2, HVT, vaccine A, and positive control groups were 0%, 100%, 60%, and 100%, respectively, and the BBIX values were 1.07 ± 0.22, 0.27 ± 0.05, 0.64 ± 0.33, and 0.32 ± 0.06, respectively. These results indicate that rHVT-VP2 can provide 100% protection against a challenge with the IBDV (G2d), whereas vaccine A only provides partial protection. In conclusion, vaccination with the recombinant virus rHVT-VP2 can provide chickens with effective protection against variant IBDV (G2d). [ABSTRACT FROM AUTHOR]

Simple Summary: Wild animals can be of veterinary and public health concern due to their potential to harbor pathogens and transmit them to humans and domestic animals. Avian paramyxovirus 1 (APMV-1) is capable of infecting a wide range of avian species, leading to a broad spectrum of clinical symptoms and posing a potential risk to public health. This study represents the investigation of avian paramyxoviruses (APMVs) in wild waterfowl from Korea. In this study, 13 isolates of APMV-1 and 1 isolate of APMV-13 were obtained and their genetic characteristics of fusion gene were analyzed. We identified the wild ducks and geese likely to be infected with APMV, and our data suggest a possible intercontinental transmission of APMVs by wild waterfowl. Moreover, our study provides the first evidence for the presence of class I of APMV-1 and APMV-13 in wild waterfowl that were sampled in Korea. This finding not only adds to our understanding of the diversity of APMVs but also underscores the importance of monitoring and addressing the epidemiology of these viruses in wildlife, particularly from a One Health perspective. Avian paramyxoviruses (APMVs) are often carried by wild waterfowl, and the wild waterfowl may play an important role in the maintenance and spread of these viruses. In this study, we investigated APMVs in the population of migratory wild waterfowl from 2015 to 2021 in Korea and analyzed their genetic characteristics. Fourteen viruses were isolated and subsequently identified as APMV-1 (n = 13) and APMV-13 (n = 1). Phylogenetic analysis of the full fusion gene of 13 APMV-1 isolates showed that 10 APMV-1 isolates belonged to the class II sub-genotype I.2, which was epidemiologically linked to viruses from the Eurasian continent, and 3 viruses belonged to class I, which linked to viruses from the USA. The APMV-13 isolates from wild geese in this study were highly homology to the virus isolated from China. Sequence analysis of 14 isolates showed that all isolates had a typical lentogenic motif at the cleavage site. In summary, we identified the wild species likely to be infected with APMV and our data suggest possible intercontinental transmission of APMV by wild waterfowl. Our current study also provides the first evidence for the presence of class I of APMV-1 and APMV-13 in wild waterfowl surveyed in Korea. [ABSTRACT FROM AUTHOR]

Metorchis orientalis is a neglected zoonotic parasite of the gallbladder and bile duct of poultry, mammals, and humans. It has been widely reported in Asian, including China, Japanese, and Korea, where it is a potential threat to public health. Despite its significance as an animal and human pathogen, there are few published transcriptomic and proteomics data available. Transcriptome Illumina RNA sequencing and label-free protein quantification were performed to compare the gene and protein expression of adult and metacercariae-stage M. orientalis , resulting in 100,234 unigenes and 3,530 proteins. Of these, 13,823 differentially expressed genes and 1,445 differentially expressed proteins were identified in adult versus metacercariae. In total, 570 genes were differentially expressed consistent with the mRNA and protein level in the adult versus metacercariae stage. Differential gene transcription analyses revealed 34,228 genes to be expressed in both stages, whereas 66,006 genes showed stage-specific expression. Compared with adults, the metacercariae stage was highly transcriptional. GO and KEGG analyses based on transcriptome and proteome revealed numerous up-regulated genes in adult M. orientalis related to microtubule-based processes, microtubule motor activity, and nucleocytoplasmic transport. The up-regulated genes in metacercariae M. orientalis were mainly related to transmembrane receptor protein serine/threonine kinase activity, transmembrane receptor protein serine/threonine kinase signaling pathway. Transcriptome and proteome comparative analyses showed numerous up-regulated genes in adult stage were mainly enriched in actin filament capping, spectrin, and glucose metabolic process, while up-regulated genes in metacercariae stage were mainly related to cilium assembly, cilium movement, and motile cilium. These results highlight changes in protein and gene functions during the development of metacercariae into adults, and provided evidence for the mechanisms involved in morphological and metabolic changes at both the protein and gene levels. Interestingly, many genes had been proved associated with liver fibrosis and carcinogenic factors were identified highly expressed in adult M. orientalis , which suggests that M. orientalis is a neglected trematode with potential carcinogenic implications. These data provide attractive targets for the development of therapeutic or diagnostic interventions for controlling M. orientalis. [ABSTRACT FROM AUTHOR]

Simple Summary: Salmonella enterica serovar Gallinarum (S. enterica ser. Gallinarum) is a host-specific agent of fowl typhoid (FT). This is one of the most important bacterial infections in the poultry industry in both developing and developed countries, including South Korea. The use of antimicrobial drugs is the first choice for disease control. Antimicrobials, such as β-lactams, aminoglycosides, and fluoroquinolones, are frequently used to treat FT. However, the continuous use of antimicrobial drugs has led to the emergence and persistence of antimicrobial-resistant S. enterica ser. Gallinarum. In this study, we analyzed the antimicrobial susceptibility and epidemiological relationship of thirty isolates of S. enterica ser. Gallinarum isolated from poultry farms with an FT outbreak from 2013 to 2018 in South Korea. All the isolates showed a multi-drug resistant (MDR) phenotype. This study confirmed horizontal transmission and cross-contamination between farms within the same integrated poultry company or between farms belonging to different companies. The characterization of these isolates would be helpful to develop prevention and control strategies for the MDR S. enterica ser. Gallinarum infection in South Korea. Antimicrobial resistance and pulsed-field gel electrophoresis (PFGE) genotypes of collected S. enterica ser. Gallinarum isolates were investigated to examine the epidemiological relationship between field outbreak isolates of S. enterica ser. Gallinarum. Thirty S. enterica ser. Gallinarum isolates collected from poultry farms with FT outbreaks from 2013 to 2018 in South Korea were analyzed. All isolates were resistant to at least 3 of the 18 antimicrobials tested and exhibited an MDR phenotype. All isolates showed resistance to streptomycin, sulfisoxazole, and colistin. One isolate was resistant to 9 antimicrobials. The antimicrobial resistance profile, streptomycin-sulfisoxazole-colistin-nalidixic acid-ciprofloxacin-gentamicin (18/30, 60.0%), was the most prevalent. PFGE types were classified into 10 groups with a 100% correlation cutoff in dendrograms for 30 field isolates. The dominant PFGE types were 1 (8/30, 26.7%), 4 (7/30, 23.3%), and 9 (5/30, 16.7%). Interestingly some isolates collected from the same and different companies had the same PFGE type. We reported a high MDR rate in S. enterica ser. Gallinarum isolates. The present study highlights the occurrence of horizontal spread and cyclic contamination of MDR S. enterica ser. Gallinarum within the same company. Furthermore, we showed cross-contamination between different companies. The characterization of these isolates would be helpful in the development of prevention and control strategies for MDR S. enterica ser. Gallinarum infection in South Korea. [ABSTRACT FROM AUTHOR]

Simple Summary: Extended-spectrum cephalosporin (ESC)-resistant Enterobacteriaceae frequently detected in humans and food-producing animals are of major concern in public health. This study was undertaken to investigate the contamination of ESC-resistant E. coli in the environment of a slaughterhouse during chicken meat processing. This study indicates that cross-contamination of ESBL/AmpC-producing E. coli has a crucial impact on the occurrence of ESC resistance in retail chicken meat. Thus, ESBL-/AmpC-producing E. coli were brought into the slaughterhouse by certain broiler chicken flocks, and other chicken flocks were contaminated by ESBL/AmpC-producing E. coli already present in the slaughterhouse environment. These findings support the hypothesis that world widely epidemic conjugative plasmids have contributed to the dissemination of ESBL/AmpC resistance in broiler chickens in Korea. As conjugative plasmids always carrying multiple resistance genes, continuous persistence of blaCTX-M and blaCMY genes located on plasmids within microbial communities will be mediated by co-selection processes with other resistance genes. Hence, further research on the control of bacterial conjugation is urgently required. Our study emphasizes that chicken slaughterhouses could perform the functions of convergence and dispersion of ESBL/AmpC resistance, and that world widely epidemic conjugative plasmids contribute to the dissemination of ESBL/AmpC from chickens to humans along the food chain. ESC-resistant E. coli isolates were collected from broiler chickens, a slaughterhouse, and retail meat to assess their dispersion and their involvement in cross-contamination. ESBL-/AmpC-producing E. coli were isolated during the slaughter process of all six investigated chicken flocks from scalding, feather removal, first conveyor, evisceration, second washing, third conveyor, and third washing areas, and from handling workers in the slaughterhouse. ESC-resistant E. coli isolates with the same pulsed-field gel electrophoresis type were found in the same site (scalding) on different sampling days. ESBL/AmpC-producing E. coli isolates were absent in the lairage area in the slaughterhouse, but present in the retail markets in 36.8% (7/19) of the chicken flocks. The blaCTX-M genes and blaCMY-2 were conjugated to recipient E. coli J53 in 67.5% (27/40) and 56.1% (23/41) of ESBL-producing and AmpC-producing E. coli isolates, respectively. The presence of the same conjugative plasmids was found in genetic diversity ESC-resistant E. coli colonies collected on different sampling days. Our study emphasizes that cross-contamination of ESBL/AmpC-producing E. coli in slaughterhouse has a crucial impact on the occurrence of ESC resistance in retail chicken meat. [ABSTRACT FROM AUTHOR]

Simple Summary: The prevalence of extended-spectrum cephalosporin (ESC)-resistant Salmonella is of great concern, as these strains with the same β-lactamase (bla) genes were found in human and poultry. The objective is to characterize ESC-resistant Salmonella isolated from chicken and to determine the transferability of β-lactamase gene-harboring plasmid in vitro and in vivo. ESC resistance genes in Salmonella isolated from chickens and presented a comprehensive analysis of the highly frequent transfer of the blaCMY-2 gene in vitro and in vivo. In addition, this study has demonstrated the ease with which a blaCMY-2 gene-harboring plasmid can be rapidly transferred between Salmonella and pathogenic E. coli within the intestinal tracts of mice, even without antimicrobial selective pressure. Given the potential risk of the frequent transfer of the blaCMY-2 gene via the food chain to the human digestive tract, the molecular mechanism involved in the dissemination and maintenance of ESC resistance genes should be studied as further research in greater detail, and enhanced surveillance should be implemented to prevent the widespread of ESC resistant strains. A total of 136 Salmonella isolates from chicken feces and meat samples of the top 12 integrated chicken production companies throughout Korea were collected. Among the 17 ESC-resistant Salmonella; blaCTX-M-15 was the most prevalent gene and two strains carried blaTEM-1/blaCTX-M-15 and blaCMY-2, respectively. The transferable blaCTX-M-15 gene was carried by IncFII plasmid in three isolates and the blaCMY-2 gene carried by IncI1 plasmid in one isolate. blaCMY-2 gene-harboring strain was selected as the donor based on the high frequency of blaCMY-2 gene transfer in vitro and its transfer frequencies were determined at 10−3 transconjugants per recipient. The transfer of blaCMY-2 gene-harboring plasmid derived from chicken isolate into a human pathogen; enteroinvasive Escherichia coli (EIEC), presented in mouse intestine with about 10−1 transfer frequency without selective pressure. From the competition experiment; blaCMY-2 gene-harboring transconjugant showed variable fitness burden depends on the parent strains. Our study demonstrated direct evidence that the blaCMY-2 gene harboring Salmonella from chicken could frequently transfer its ESC-resistant gene to E. coli in a mouse intestine without antimicrobial pressure; resulting in the emergence of multidrug resistance in potentially virulent EIEC isolates of significance to human health; which can increase the risk of therapeutic inadequacy or failures. [ABSTRACT FROM AUTHOR]

Simple Summary: Investigation of Campylobacter prevalence throughout the entire chicken production process from farms to retail meat is still limited. In this study, we examined the prevalence and antimicrobial susceptibility of Campylobacter in 10 production lines from one of the largest integrated poultry production companies in Korea. The prevalence of Campylobacter in breeder farm, hatchery, broiler farm, slaughterhouse, and retail meat products was 50.0%, 0%, 3.3%, 13.4%, and 68.4%, respectively. Resistance to fluoroquinolones was the most frequently observed, and 16 isolates from breeder farm were resistant to both azithromycin and ciprofloxacin. Diverse pulsed-field gel electrophoresis genotypes were presented with discontinuous patterns along the whole production chain. Thirty percent of Campylobacter-free flocks became positive after slaughtering. An identical genotype was simultaneously detected from both breeder farm and retail meat, even from different production lines. This study reveals that antimicrobial-resistant Campylobacter contamination can occur at all stages of the chicken supply chain. In particular, the breeder farm and slaughterhouse should be the main control points, as they are the potential stages at which antimicrobial-resistant Campylobacter could spread to retail meat products by horizontal transmission. The aim of this study was to analyze the prevalence, antimicrobial resistance, and genetic diversity of Campylobacter isolates that were obtained from whole chicken production stages in Korea. A total of 1348 samples were collected from 10 production lines. The prevalence of Campylobacter in breeder farm, broiler farm, slaughterhouse, and retail meat products was 50.0%, 3.3%, 13.4%, and 68.4%, respectively, and Campylobacter was not detected at the hatchery stage. Resistance to quinolones/fluoroquinolones was the most prevalent at all stages. Among the multidrug-resistant isolates, 16 isolates (19.8%) from breeder farm were resistant to both azithromycin and ciprofloxacin. A total of 182 isolates were subdivided into 82 pulsed-field gel electrophoresis (PFGE) genotypes with 100% similarity. Diverse genotypes were presented with discontinuous patterns along the whole production chain. Thirty percent of Campylobacter-free flocks became positive after slaughtering. An identical genotype was simultaneously detected from both breeder farm and retail meat, even from different production lines. This study reveals that antimicrobial-resistant Campylobacter contamination can occur at all stages of the chicken supply chain. In particular, the breeder farm and slaughterhouse should be the main control points, as they are the potential stages at which antimicrobial-resistant Campylobacter could spread to retail meat products by horizontal transmission. [ABSTRACT FROM AUTHOR]

Simple Summary: The present study investigated the emergence, antimicrobial susceptibility profiles, serotypes, and genotypes of Salmonellaenterica from hatcheries and their upstream breeder farms to determine the occurrence of antimicrobial-resistant (AMR) Salmonellaenterica contamination in hatcheries and its dissemination in an integrated broiler chicken operation. The hatcheries showed a high prevalence of Salmonella isolates with high antimicrobial resistance and no susceptible isolate. The AMR isolates from hatcheries originating from breeder farms could disseminate to the final retail market along the broiler chicken supply chain. Hatcheries play a more important role than breeder farms in the occurrence and spread of Salmonella in the broiler production chain. The emergence of AMR Salmonella in hatcheries may be due to the horizontal spread of resistant isolates rather than the first contamination of susceptible Salmonella and acquisition of resistance. Therefore, Salmonella control in hatcheries, particularly its horizontal transmission, is important. Positive identification rates of Salmonella enterica in hatcheries and upstream breeder farms were 16.4% (36/220) and 3.0% (6/200), respectively. Among the Salmonella serovars identified in the hatcheries, S. enterica ser. Albany (17/36, 47.2%) was the most prevalent, followed by the serovars S. enterica ser. Montevideo (11/36, 30.6%) and S. enterica ser. Senftenberg (5/36, 13.9%), which were also predominant. Thirty-six isolates showed resistance to at least one antimicrobial tested, of which 52.8% (n = 19) were multidrug resistant (MDR). Thirty-three isolates (enrofloxacin, MIC ≥ 0.25) showed point mutations in the gyrA and parC genes. One isolate, S. enterica ser. Virchow, carrying the blaCTX-M-15 gene from the breeder farm was ceftiofur resistant. Pulsed-field gel electrophoresis (PFGE) showed that 52.0% S. enterica ser. Montevideo and 29.6% S. enterica ser. Albany isolates sourced from the downstream of hatcheries along the broiler chicken supply chain carried the same PFGE types as those of the hatcheries. Thus, the hatcheries showed a high prevalence of Salmonella isolates with high antimicrobial resistance and no susceptible isolate. The AMR isolates from hatcheries originating from breeder farms could disseminate to the final retail market along the broiler chicken supply chain. The emergence of AMR Salmonella in hatcheries may be due to the horizontal spread of resistant isolates. Therefore, Salmonella control in hatcheries, particularly its horizontal transmission, is important. [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate variation in antimicrobial resistance in Clostridium perfringens (C. perfringens) isolated from chickens after withdrawal of antimicrobial growth promoters (AGPs); and to investigate the correlation between the presence of toxin genes (cpb2, netB, and tpeL) and antimicrobial resistance. Altogether, 162 isolates of C. perfringens were obtained from chickens displaying clinical signs of necrotic enteritis (n = 65) and from healthy chickens (n = 97) in Korea during 2010–2016. Compared to before AGP withdrawal, increased antimicrobial resistance or MIC50/MIC90 value was observed for nine antimicrobials including penicillin, tetracycline, tylosin, erythromycin, florfenicol, enrofloxacin, monensin, salinomycin, and maduramycin. Significantly (p < 0.05) higher resistance to gentamicin, clindamycin, and virginiamycin was found in isolates from chickens with necrotic enteritis compared to those from healthy chickens. tpeL gene was not detected in C. perfringens isolates from healthy chickens. A correlation between toxin gene prevalence and antibiotic resistance was found in the C. perfringens isolates. Because the usage of antimicrobials may contribute to the selection of both resistance and toxin genes, these can potentially make it challenging to control antimicrobial resistance in pathogenic colonies. Therefore, a more complete understanding of the interplay between resistance and virulence genes is required. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the prevalence, serovar distribution, antimicrobial resistance, and genotypic analyses of the dominating serovars of Salmonella in chickens from a national study in Korea. Between 2017 and 2018, a total of 550 chicken samples were collected from the top 12 integrated broiler chicken operations in Korea. Salmonella was isolated from 117 (32.5%) chicken feces and 19 (10.0%) retail chicken meat sources. Ten serovars were identified, and the most common Salmonella serovar was Salmonella ser. Albany (50 isolates, 36.8%), followed by S. Enteritidis (38 isolates, 27.9%), and S. Montevideo (23 isolates, 16.9%) isolated from 6, 10, and 6 operations, respectively. A total of 35 (25.7%) isolates were with the ACSSuTN (ampicillin, chloramphenicol, streptomycin, sulfisoxazole, tetracycline, and nalidixic acid) resistance pattern, with high prevalence of this resistance pattern in S. Albany (29 isolates, 58.0%). A total of 35 PFGE types were identified among Salmonella isolates of the serovars Albany, Enteritidis, Virchow, Montevideo, and Senftenberg, while 11 distinct types of PFGE patterns were found among S. Albany isolates, which showed an overall homology similarity of higher than 85%. Among these 35 PFGE types, 22 PFGE types corresponded to 32 isolates from samples limited to one operation, and the other 13 PFGE types corresponded to 72 isolates from samples widely distributed among different operations. These results highlighted rapid colony dissemination of multidrug-resistant S. Albany in chicken all over Korea after it first appeared in 2016; furthermore, the spread of Salmonella colonies between various integrated operations was common, and several operations played an important role in Salmonella carriage and transmission in Korea. [ABSTRACT FROM AUTHOR]

• First report of Salmonella enterica in migratory birds in South Korea. • The prevalence (0.93 %) of Salmonella , comprised S. Typhimurium, Berta, and Virchow. • Migratory birds serve as reservoirs for antimicrobial resistant Salmonella. • Dissemination of Salmonella isolates with multiple virulence genes across migratory birds. • Identical ciprofloxacin resistant colony was found in migratory birds and chickens. Salmonella is a type of zoonotic bacteria that represents an economic and public health concern worldwide. Difficulties in sample collection from migratory birds mean little is known about their importance as a reservoir of Salmonella. The present study evaluated the prevalence and potential risk of Salmonella enterica in migratory birds. From 2012–2017, 3661 cloacal swabs from migratory birds were collected in South Korea and tested to isolate S. enterica. Strains were tested for antimicrobial resistance and the presence of virulence genes. Thirty-six S. enterica strains, including S. enterica serovar Typhimurium (n = 19), S. Berta (n = 16), and S. Virchow (n = 1), were isolated from 34 birds. Two migratory birds were simultaneously co-infected with two serotypes. S. enterica was isolated from the Mallard duck, Northern pintail, Eurasian wigeon, Spot-billed duck, Eastern great egret, and Intermediate egret. S. Virchow was resistant to ciprofloxacin, with a point mutation (Ser-83-Phe) in the gyr A gene. Ten virulence genes were detected; sixteen strains were positive for all ten virulence genes. Salmonella was isolated from different migratory bird species and geographic locations with up to 100 % similarity of PFGE type. Eight S. Virchow strains taken from migratory birds, poultry farms, and chicken meat showed the same PFGE type. Salmonella was transmitted across species, space, and time in migratory birds. These birds may play a role in the dispersal of pathogenic and antimicrobial-resistant Salmonella and sporadic Salmonella infections in poultry; therefore, they may represent a direct or indirect public health threat. [ABSTRACT FROM AUTHOR]