Huong Thi Thanh Doan

Classical swine fever is one of the most dangerous infectious diseases, causing significant economic damage to the pig industry globally and in Vietnam. The disease is caused by the Classical Swine Fever Virus (CSFV), an RNA virus with a high mutation rate. Classical swine fever remains a complex and persistent threat. In this study, the full E2 gene of CSFV strains collected from Ha Noi (21.028333 N, 105.853333 E), Da Nang (16.079 N, 108.217 E), and Quang Ninh (21.250982 N, 107.193604 E) was analyzed alongside three vaccine strains currently used in Vietnam. The results show that the highly virulent CSFV strains circulating in 2023 belong to genotypes 1.1, 2.1, and 2.2, while the vaccine strains in use belong to genotype 1.1. There are notable differences in amino acids at key antigen epitope sites between the vaccine strains and the circulating CSFV strains in 2023. These findings highlight the need to develop new vaccines that are better suited to the currently circulating virus strains.

Infectious bronchitis is an acute disease in chickens caused by the Infectious Bronchitis Virus (IBV). The disease causes serious economic losses in many countries in the world that have chicken farming industries. Even though vaccines have been used, the disease is still very complicated in many provinces and cities in Vietnam. The reason is that the virus strains causing the disease in the field have many changes compared to the vaccine virus strains. Research showed that there are numerous strains and genotypes of IBV in Vietnam. However, only the S1 gene has been studied, and no research has been done on the full genome. In this study, the IBV strain causing disease in 2021 in Hung Yen Province (North Vietnam) was sequenced and analyzed for molecular characteristics. The analysis results showed that strain IBHYM belongs to genotype QX-like (GI-19) and has a close relationship with Chinese IBV strains. The IBHYM strain has higher nucleotide and amino acid similarities with strains of genotype QX-like (87.5% to 95% for nucleotide and 80.2% to 91.3% for amino acid) and the lowest similarity to strains of genotype TC07-like (70.4% to 70.5% for nucleotide and 53.8% to 56.2% for amino acid). The field pathogenic virus strains also have low nucleotide and amino acid similarity with the Massachusetts vaccine strain currently in use (86% for nucleotide and 77.1% for amino acid). Research results show the importance of molecular epidemiological surveillance and the selection of appropriate vaccine strains in Vietnam. This is the first report of sequencing the whole IBV genome in Vietnam.

Infectious bronchitis virus (IBV) is considered as one of the main causes of economic loss in chicken farms worldwide, especially in poultry producing nations. This virus, a member of the Coronaviridae family, is classified into different genotypes based on its surface spike glycoproteins which also play important roles in cell attachment and immune response. Due to continual genetic mutation of IBV, attenuated and inactivated vaccines show decreased effectiveness or lack of cross – protection; thus ongoing studies focus on the development of new generation vaccines to prevent new IBV outbreaks. This paper describes our study on expression of the antigenic region in S1 subunit of the spike in Nicotiana benthamiana. Two expression vectors carrying either S1 or receptor binding domain (RBD) coding gene (pCB301-S1 and pCB301-RBD) were constructed and transformed into tobacco leaves by agroinfiltration method. The RBD showed a clearly higher level of expression compared to the whole S1. Purification of RBD by immobilized metal ion chromatography and size exclusion chromatography represented a mixture of monomer, dimer and trimer glycoprotein with expected sizes in Western blot. In summary, this study demonstrated our primary success in establishing an expression model that could be used to investigate plant-based IBV recombinant vaccine.

We conducted nucleotide and amino acid sequence alignment and phylogenetic analysis of porcine circovirus ORF2 (Cap protein) from 17 PCV2-positive clinical samples from nine different northern Vietnamese provinces (Mar 2018–Nov 2020), four local vaccines, and 77 reference strains. We identified one PCV2a (1/17 = 5.9%), five PCV2b (5/17 = 29.9%), and 11 PCV2d (11/17 = 64.7%) isolates, while only PCV2d was detected in 2020. Timeline analysis indicated an increasing predominance of PCV2d nationwide (2018–2020). With strong nodal support (98% for nucleotides and 74% for amino acids), the phylogenetic tree topology revealed a distinct PCV2h clade including recombinant/intermediate strains and local vaccines. The Cap protein sequences from 11 PCV2d field strains had the 2d-genotype-typical motif ⁸⁶SNPLSV⁹¹ in loop CD, the motif TGID in loop GH-HI, and the motif ²³⁰PLNPK²³⁴ in loop CT. The PCV2h isolates (and vaccines) had the ⁸⁶SNPLSV⁹¹, SAID, and ²³⁰L(N/H)PK²³⁴ motifs. Selection pressure analysis indicated positive selection at seven sites: A⁶⁸N in immunoreactive region (IRR)-A; ¹¹⁹G and ¹³⁰V in IRR-B; and ¹⁶⁷L, T¹⁹⁰(A/S), ¹⁹⁴D and ²⁰²F in IRR-C. We identified PCV2h as the genotype of the recombinant strains, which resulted from intergenotype recombination of PCV2a, PCV2b, and PCV2d. The current data provide new information about the diversity, distribution, and dominance of the PCV2 genotype in Vietnam.

Viral protein 2 (VP2) of canine parvovirus (CPV) exhibits a high degree of genetic and antigenic diversity. We analyzed 88 Vietnamese CPV-VP2 sequences (1755 bp), 34 from this study and 54 from previous studies, and discovered a new sublineage, “new var.”, within the lineage CPV-2c-“new”, characterized by the mutation 5G/447M, which is restricted to the Vietnamese isolates. These new mutants appear to have emerged in recent years, accounting for 65.5% of the total. With strong nodal support (98%), the distinct Vietnamese 2c-“new-var.” sublineage (5G/426E/447M) was found to be separate from the 2c-“new” sublineage (5G/426E/447I) within the 2c-(Asia)/Asia-2c lineage. Amino acid changes in epitopes of VP2 might have led to the generation of subvariants and affected the antigenicity, immunogenicity, or virulence of the virus, resulting in vaccine failure worldwide.

Heterophyidiasis is now a major public health threat in many tropical countries. Species in the trematode family Heterophyidae infecting humans include Centrocestus formosanus, Haplorchis pumilio, H. taichui, H. yokogawai, Procerovum varium and Stellantchasmus falcatus. For molecular phylogenetic and systematic studies on trematodes, we need more prospective markers for taxonomic identification and classification. This study provides near-complete ribosomal transcription units (rTU) from Haplorchis pumilio and H. taichui and demonstrates the use of 28S rDNA sequences for identification and phylogenetic analysis. The near-complete ribosomal transcription units (rTU), consisting of 18S, ITS1, 5.8S, ITS2 and 28S rRNA genes and spacers, from H. pumilio and H. taichui from human hosts in Vietnam, were determined and annotated. Sequence analysis revealed tandem repetitive elements in ITS1 in H. pumilio and in ITS2 in H. taichui. A phylogenetic tree inferred from 28S rDNA sequences of 40 trematode strains/species, including 14 Vietnamese heterophyid individuals, clearly confirmed the status of each of the Vietnamese species: Centrocestus formosanus, Haplorchis pumilio, H. taichui, H. yokogawai, Procerovum varium and Stellantchasmus falcatus. However, the family Heterophyidae was clearly not monophyletic, with some genera apparently allied with other families within the superfamily Opisthorchioidea (i.e. Cryptogonimidae and Opisthorchiidae). These families and their constituent genera require substantial re-evaluation using a combination of morphological and molecular data. Our new molecular data will assist in such studies. The 28S rDNA sequences are conserved among individuals within a species but varied between genera. Based on analysis of 40 28S rDNA sequences representing 19 species in the superfamily Opisthorchioidea and an outgroup taxon (Alaria alata, family Diplostomidae), six common human pathogenic heterophyids were identified and clearly resolved. The phylogenetic tree inferred from these sequences again confirmed anomalies in molecular placement of some members of the family Heterophyidae and demonstrates the need for reappraisal of the entire superfamily Opisthorchioidea. The new sequences provided here supplement those already available in public databases and add to the array of molecular tools that can be used for the diagnosis of heterophyid species in human and animal infections.

Introduction: The aim of this study was to identify the genetic characteristics and molecular genotyping of duck hepatitis A virus (DHAV) isolated in Vietnam during 2009–2013. Methodology: Thirty duckling livers from outbreaks between 2009 and 2013 in seven provinces were collected and identified by polymerase chain reaction (PCR). Then, VP1 genes of eleven positive samples and two attenuated vaccine strains were sequenced and analyzed. Results: Genotypic and phylogenetic analyses indicated that the 13 Vietnamese isolates were classified into two genotypes, DHAV-1 and DHAV-3. The rate of identity and homology was 91%–100% between the 10 Vietnamese and 26 global strains of DHAV-3, and 92%–100% between 3 Vietnamese and 16 strains of DHAV-1. Between the DHAV-3 and DHAV-1 strains, the divergence reached 30%. At the C-terminal of VP1 for the different strains, a hypervariable region was observed, and notably, six of the Vietnamese DHAV-3 strains in this study showed four consistent differences (at positions T184M, Q200H, K207N, and K214R) within this group that were distinct from all other DHAV-3 strains. Conclusions: This is the first report of molecular characterization of DHAVs in Vietnam. At least two genotypes were identified, DHAV-1 and DHAV-3, with diversified clades within and between genotypes. DHAV-3 seemed to be dominant in Vietnam.