Complete Genome Sequence Analysis Research Articles

Molecular and Genetics-Based Systems for Tracing the Evolution and Exploring the Mechanisms of Human Norovirus Infections.

Human noroviruses (HuNoV) are major causes of acute gastroenteritis around the world. The high mutation rate and recombination potential of noroviruses are significant challenges in studying the genetic diversity and evolution pattern of novel strains. In this review, we describe recent advances in the development of technologies for not only the detection but also the analysis of complete genome sequences of noroviruses and the future prospects of detection methods for tracing the evolution and genetic diversity of human noroviruses. The mechanisms of HuNoV infection and the development of antiviral drugs have been hampered by failure to develop the infectious virus in a cell model. However, recent studies have demonstrated the potential of reverse genetics for the recovery and generation of infectious viral particles, suggesting the utility of this genetics-based system as an alternative for studying the mechanisms of viral infection, such as cell entry and replication.

The genetic and biological characterization of the first avian paramyxovirus serotype 14 isolated from chicken in China.

In October 2020, an avian paramyxovirus serotype 14 (APMV-14)-designated chicken/Fujian/2160/2020 (FJ2160) was isolated from tracheal and cloacal swab sample of chicken collected from live bird market in Fujian province in China during the active surveillance program. The complete genome of FJ2160 comprised 15,444 nucleotides (nt) complying with the paramyxovirus "rule of six" and encoded six non-overlapping structural proteins in the order of 3'-NP-P-M-F-HN-L-'5. The complete genome sequence analysis showed that FJ2160 had the highest identity (90.0%) with the APMV-14 isolated from Japan, while the nucleotide sequence identities of FJ2160 and other APMVs ranged from 42.4 to 51.1%. The F protein cleavage site was TREGR↓L, which resembled a lentogenic strain of APMV-1. Phylogenetic analysis revealed that the FJ2160 closest relative was APMV-14. The intracerebral pathogenicity index (ICPI) tests indicated that the virus was lentogenic. This is the first report of APMV-14 in China. These results provide evidence that APMV-14 could infect chickens and reveal the genetic characteristics and biological properties of the virus, which can help to better understand this new emerging APMV-14.

Complete genome sequence of a novel closterovirus isolated from Dregea volubilis.

The complete genome sequence of a putative novel closterovirus, tentatively named "Dregea volubilis closterovirus 1" (DvCV1, GenBank accession no. MZ779122), infecting Dregea volubilis in China was determined using high-throughput sequencing (HTS). The complete genome sequence of DvCV1 consists of 16,165 nucleotides (nt) and contains nine ORFs. The genome structure of DvCV1 is typical of members of the genus Closterovirus. Complete genome sequence analysis showed that DvCV1 shares 41.4-48.4% nucleotide sequence identity with other known closteroviruses. The putative RNA-dependent RNA polymerase (RdRp), heat shock protein 70-like protein (HSP70h), and coat protein (CP) of DvCV1 share 46.80-62.65%, 31.06-51.80%, and 28.34-37.37% amino acid sequence identity, respectively, with the RdRp, HSP70h and CP of other closteroviruses. Phylogenetic analysis based on HSP70h aa sequences placed DvCV1 alongside other members of the genus Closterovirus in the family Closteroviridae. These results suggest that DvCV1 is a new member of the genus Closterovirus. This is the first report of a closterovirus infecting D. volubilis.

Complete genome sequence analysis, morphology and structural protein identification of two Bacillus subtilis phages, BSTP4 and BSTP6, which may form a new species in the genus Salasvirus.

In the present study, two new Bacillus subtilis phages, BSTP4 and BSTP6, were isolated and studied further. Morphologically, BSTP4 and BSTP6 are podoviruses with complete genome of 19,145 (39.9% G + C content) and 19,367bp (39.8% G + C content), respectively, which became among the smallest Bacillus phages. Three most prominent structural proteins were separated and identified as pre-neck appendage, major head, and head fiber proteins using LC-MS/MS. Both phages encode putative terminal proteins (TP) and contain short inverted terminal repeats (ITRs) which may be important for their replication. In addition, non-coding RNA (pRNA) and parS sites were identified which may be required for DNA packaging and their maintenance inside the host, respectively. Furthermore, the phage genome sequences show significant similarity to B. subtilis group species genome sequences. Finally, phylogenomic and phylogenetic analyses suggest that BSTP4 and BSTP6 may form a new species in the genus Salasvirus, subfamily Picovirinae of family Salasmaviridae. Considering the small numbers of ICTV-accepted B. subtilis phages and the importance of the host in the food industry and biotechnology, the current study helps to improve our understanding of the diversity of B. subtilis phages and shed light on the phage-host relationships.

Complete genome sequence and pathogenicity analysis of a highly pathogenic FAdV-4 strain

Fowl adenovirus serotype 4 (FAdV-4) is a double-stranded DNA virus that mainly infects broiler chickens and has caused huge economic losses to the poultry industry. Recently, an FAdV-4 strain, SDLC202009, the causative pathogen of hydropericardium-hepatitis syndrome (HHS) in Liaocheng, Shandong, was isolated from commercial laying hens and propagated in specific pathogen free SPF chicken embryos. Pathogenicity studies showed that SDLC202009 could infect SPF chicken embryos and chickens, with a mortality rate of 100%. The complete genome was sequenced, and phylogenetic analysis showed that SDLC202009 belonged to the FAdV-4 cluster, with a genome length of 43, 077 bp. The SDLC202009 had 99.9% identity with the JSJ13 and SD1601, which were recently isolated in China. Compared to the recently isolated strain in China, SDLC202009 had deleted open reading frame 19 (ORF19), ORF27, ORF48, and ORF0. SDLC202009 harbored amino acid site mutations in the main structural proteins hexon, fiber1, and fiber2 similar with those in highly pathogenic strains. Furthermore, SDLC202009 showed unique mutations in hexon A571P, fiber1 E216K, and fiber2 N98K. In summary, our findings provide theoretical support for prevention and control of the HHS.

Complete genome sequence analysis of a novel citlodavirus isolated from the leaves of Myrica rubra in Yunnan.

Through high-throughput sequencing, a novel citlodavirus, tentatively named "Myrica rubra citlodavirus 1" (MRV1,accession no. OP374189), was isolated from the leaves of Myrica rubra in Yunnan exhibiting narrow deformity of leaf tips, shrinkage, and chlorosis along the veins. The complete genome sequence was determined and analyzed using cloning and Sanger sequencing. MRV1 is a single-stranded circular non-enveloped DNA virus with a genome size of 3775 nucleotides and contains six open reading frames (ORFs). The virion-sense genome strand encodes a coat protein (CP, nt 750-1,493, 247 aa), two hypothetical movement proteins (V3, nt 382-666, 94 aa; and V2, nt 461-895, 144 aa), and one movement protein (MP, nt 1,527-2,438, 303 aa). The complementary strand of the genome encodes two replication proteins (RepA, nt 3,712-2,834, 292 aa; Rep, nt 2,867-2,553, 104 aa). The MRV1 genome contains the stem-loop motif 5'-TAATATTAC-3', which is a highly conserved nonanucleotide motif found in the origin of virion-strand replication in geminiviruses. Genome sequence alignment analysis showed that citrus chlorotic dwarf associated virus (CCDaV, accession no. JQ920490) shared the highest nucleotide sequence similarity (66.10% identity) with MRV1. Phylogenetic analysis showed that CCDaV is the closest known relative of MRV1, and that these viruses clustered in a single branch within a clade consisting of citlodaviruses. These results indicate that MRV1 should be regarded as a new species of the genus Citlodavirus in the family Geminiviridae.

Non-structural genes of novel lemur adenoviruses reveal codivergence of virus and host.

Adenoviruses (AdVs) are important human and animal pathogens and are frequently used as vectors for gene therapy and vaccine delivery. Surprisingly, there are only scant data regarding primate AdV origin and evolution, especially in the most basal primate hosts. We detect and sequence AdVs from faeces of two Madagascan lemur species. Complete genome sequence analyses define a new AdV species with a particularly large gene encoding a protein of unknown function in the early gene region 3. Unexpectedly, the new AdV species is not most similar to human or other simian AdVs but to bat adenovirus C. Genome characterisation shows signals of virus-host codivergence in non-structural genes, which show lower diversity than structural genes. Outside a lemur species mixing zone, recombination less frequently separates structural genes, as in human adenovirus C. The evolutionary history of lemur AdVs likely involves both a host switch and codivergence with the lemur hosts.

Biofilm removal mediated by Salmonella phages from chicken-related sources

Salmonella and their biofilm formation are the primary bacterial causes of foodborne outbreaks and cross-contamination. The objective of the study was to investigate the potential of Salmonella phages as an alternative technology for biofilm removal. In this work, 21 Salmonella phages were isolated from a chicken farm and slaughter plant and the phage (CW1) with the broadest spectrum was characterized. Complete genome sequence analysis revealed that the genomes of phage CW1 is composed of 41763 bp with 58 open reading frames (ORFs) and a holin-endolysin system and it does not encode any virulence or lysogeny. A phage cocktail consisted of CW1 (with the broadest spectrum of 70.49 %) and CW11, M4 and M10 (with a high lytic activity of more than 67.11 %) was established. Treatment with the cocktail reduced the cells in the developing biofilm and mature biofilm by 0.79 lg(CFU/cm2) and 0.4 lg(CFU/cm2), respectively. More dead cells and scattered extracellular polymeric substances (EPS) were observed by confocal laser scanning microscopy and scanning electron microscopy. Raman analysis found that carbohydrates and proteins were the identification receptors for scattered EPS. This finding suggests that this phage cocktail has potential applications for the sterilization of Salmonella biofilm during meat processing.

Complete genome sequence analysis and biological characteristics of Newcastle disease viruses from different hosts in China.

Newcastle disease (ND) is one of the most serious diseases affecting poultry worldwide. In 2022, we studied two strains of Newcastle disease virus (NDV) from pigeons and magpies identified by PCR and propagated in SPF chicken embryos. The whole genome of the virus was then expanded and its biological characteristics were studied. The results showed that NDV was isolated from pigeons and magpies. Virus present in the allantoic fluid could agglutinate red blood cells and could not be neutralized by serum positive for avian influenza. Sequencing showed that the gene length of the two isolates was 15,191bp, had high homology and was located in the same branch of the phylogenetic tree, both belonging to genotype VI.1.1. The sequence of 112-117 amino acids in the F gene sequence was 112R-R-Q-K-R-F117, which constituted virulent strain characteristics. The HN gene contained 577 amino acids, which is also consistent with the characteristics of a virulent strain. The results from the study of biological characteristics revealed that the virulence of SX/TY/Pi01/22 was slightly stronger. There were only four different bases in the complete sequence of the two strains. Comprehensive analysis revealed that the G at 11,847 site of the SX/TY/Ma01/22 strain may change to T, leading to translation of amino acids from R to S, thereby weakening viral virulence. Therefore, NDV was transmitted from pigeons to magpies, indicating that the pathogen could be transmitted between poultry and wild birds.

Complete chloroplast genome sequence and phylogenetic analysis of Rhododendron molle G. Don, an endangered Ericaceae species located on Dabie Mountains (central China)

Complete chloroplast genome sequence and phylogenetic analysis of Rhododendron molle G. Don, an endangered Ericaceae species located on Dabie Mountains (central China)

Characterization and complete genome sequence analysis of the novel phage RPZH3 infecting Ralstonia solanacearum

A novel lytic Ralstonia phage, RPZH3, was isolated from the soil of a tobacco field via a double agar overlay plaque assay. The phage has an icosahedral head 75 ± 5 nm in diameter with a short tail about 15 ± 5 nm in length. It was able to infect 18 out of 30 tested strains of R. solanacearum isolated from tobacco, sweet potato, tomato, pepper, and eggplant. The latent period of the phage was 80 min, and the burst period was 60 min with a burst size of about 27 pfu/cell. The phage was stable at pH 4-12 at 28°C, and it was also stable at temperatures from 45°C to 60°C at pH 7.0. The complete genome of phage RPZH3 consists of 65,958 bp, with a GC content of 64.93%. The genome contains 93 open reading frames (ORFs) and encodes a tRNA for cysteine. Nucleotide sequence alignment and phylogenetic analysis indicated that RPZH3 is a new member of the genus Gervaisevirus belonging to the class Caudoviricetes.

Complete genome sequence and antimicrobial resistance analysis of ESBL-producing Shigella sonnei carrying small cryptic plasmids isolated in northern Italy

Herein, we sequenced and assembled the genome of an S. sonnei isolate carrying several small plasmids using a hybrid approach combining Oxford Nanopore technologies (ONT) and Illumina platforms. Whole genome sequencing was conducted using the Illumina iSeq 100 and the Oxford Nanopore MinION systems, and the resulting reads were used for hybrid genome assembly with Unicycler. Coding sequences were annotated with RASTtk, while genes involved in antimicrobial resistance and virulence were identified with AMRFinderPlus. Plasmid nucleotide sequences were aligned to the NCBI nr database using BLAST, and replicons were identified using PlasmidFinder. The genome consisted of a chromosome of 4.801.657 base pairs, three major plasmids (212.849 bp, 86.884 bp and 83.425 bp) and 12 small cryptic plasmids (ranging from 8.390 bp to 1.822 bp). BLAST analysis revealed that all plasmids were highly similar to previously deposited sequences. Genome annotation predicted 5.522 coding regions, including 19 antimicrobial resistance genes, four of which located in small plasmids, and 17 virulence genes, with four of these located in a large virulence plasmid. The presence of antimicrobial resistance genes on small cryptic plasmids may represent an overlooked mechanism for the propagation of these genes among bacterial populations. Our work provides new data on these elements which may inform the development of new strategies to control the spread of ESBL-producing bacterial strains.

A Guide to Molecular Characterization of Genotype II African Swine Fever Virus: Essential and Alternative Genome Markers.

African swine fever is a contagious viral disease that has been spreading through Europe and Asia since its initial report from Georgia in 2007. Due to the large genome size of the causative agent, the African swine fever virus (ASFV), the molecular epidemiology, and virus evolution are analyzed by employing different markers. Most of these markers originate from single nucleotide polymorphisms or disparities in the copy number of tandem repeat sequences observed during the comparisons of full genome sequences produced from ASFVs isolated during different outbreaks. Therefore, consistent complete genome sequencing and comparative analysis of the sequence data are important to add innovative genomic markers that contribute to the delineation of ASFV phylogeny and molecular epidemiology during active circulation in the field. In this study, the molecular markers currently employed to assess the genotype II ASFVs circulating in Europe and Asia have been outlined. The application of each of these markers to differentiate between ASFVs from related outbreaks is described to implement a guideline to their suitability for analyzing new outbreaks. These markers do not signify the complete repertoire of genomic differences between ASFVs, but will be beneficial when analyzing the first outbreaks in a new region or a large number of samples. Furthermore, new markers must be determined via complete genome sequence analyses for enabling in-depth insights into the molecular epidemiology of ASFV.

First Molecular Detection and Genetic Analysis of a Novel Porcine Circovirus (Porcine Circovirus 4) in Dogs in the World.

A novel circovirus species was identified in farmed pigs and designated porcine circovirus 4 (PCV4); it has recently been proved to be pathogenic to piglets. However, little is known about its cross-species transmission, and there is no evidence of PCV4 in dogs. A total of 217 fecal samples were collected from diarrheal dogs in Henan Province, China, and tested for the presence of PCV4 using a real-time PCR assay. Among the 217 samples, the total positivity rate for PCV4 was 5.99% (13/217 samples), with rates of 7.44% and 4.17% in 2020 and 2021, respectively. PCV4 was detected in dogs in 6 of 10 cities, demonstrating that PCV4 could be detected in dogs in Henan Province, China. One PCV4 strain (HN-Dog) was sequenced in this study and shared high levels of identity (97.9% to 99.6%) with reference strains at the genome level. Phylogenetic analysis based on complete genome sequences of HN-Dog and 42 reference strains showed that the HN-Dog strain was closely related to 3 PCV4 reference strains (from pig, raccoon dog, and fox) but differed genetically from other viruses in the genus Circovirus. Three genotypes, i.e., PCV4a, PCV4b, and PCV4c, were confirmed by phylogenetic analysis of complete genome sequences of 42 PCV4 strains, and one amino acid variation in Rep protein (V239L) and three amino acid variations in Cap protein (N27S, R28G, and M212L) were considered conserved genotype-specific molecular markers. In conclusion, the present study is the first to report the discovery of the PCV4 genome in dogs, and the association between PCV4 infection and diarrhea warrants further study. IMPORTANCE This study is the first to report the presence of PCV4 in dogs worldwide, and the first complete genome sequence was obtained from a dog affected with diarrhea. Three genotypes of PCV4 strains (PCV4a, PCV4b, and PCV4c) were determined, as supported by specific amino acid markers (V239L for open reading frame 1 [ORF1] and N27S R28G and M212L for ORF2). These findings help us understand the current status of intestinal infections in pet dogs in Henan Province, China, and also prompted us to accelerate research on the pathogenesis, epidemiology, and cross-species transmission of PCV4.

Complete genome sequence analysis of a new potyvirus isolated from Paris polyphylla var. yunnanensis

The complete genome sequence of a new potyvirus from Paris polyphylla var. yunnanensis was determined. Its genomic RNA consists of 9571 nucleotides (nt), excluding the 3'-terminal poly(A) tail, containing the typical open reading frame (ORF) of potyviruses and encoding a putative large polyprotein of 3061 amino acids. The virus shares 54.20%-59.60% nt sequence identity and 51.80%-57.90% amino acid sequence identity with other potyviruses. Proteolytic cleavage sites and conserved motifs of potyviruses were identified in the polyprotein and within individual proteins. Phylogenetic analysis indicated that the virus was most closely related to lily yellow mosaic virus. The results suggest that the virus should be classified as a member of a novel species within the genus Potyvirus, and we have tentatively named this virus "Paris yunnanensis mosaic chlorotic virus" (PyMCV).

Complete genome sequence analysis of the multidrug resistant Aeromonas veronii isolated for the first time from stinging catfish (Shing fish) in Bangladesh.

Whole genome sequencing (WGS) of Aeromonas veronii Alim_AV_1000 isolated from ulcerative lesions of Shing fish (stringing catfish; Heteropneustes fossilis) was performed during the outbreak year 2021. Using next-generation sequencing (Illumina) technology, WGS was accomplished, resulting in the sequencing, assembly, and analysis of the entire genome of the A. veronii strain. Moreover, the genomic features, virulence factors, antimicrobial resistome, and phylogenetic analysis for the molecular evolution of this strain were also examined. The genome size of the A. veronii Alim_AV_1000 strain was 4,494,515 bp, with an average G+C content of 58.87%. Annotation revealed the known transporters and genes linked to virulence, drug targets, and antimicrobial resistance. The findings of the phylogenetic analysis revealed that the strain of the present study has a close relationship with the China strain TH0426 and strain B56. This study provides novel information on A. veronii isolated from Shing fish in Bangladesh.

Complete genome sequence analysis of a plant growth-promoting phylloplane Bacillus altitudinis FD48 offers mechanistic insights into priming drought stress tolerance in rice

Bacillus altitudinis FD48 is a multifunctional plant growth-promoting bacterium isolated from the phylloplane of rice and survives at ‐−10 bars of osmotic potential (‐−1.0 MPa). It also serves as an ideal PGPM against drought stress by triggering antioxidant defense mechanisms in rice. To further unravel the genetic determinants of osmotic stress tolerance and plant growth-promoting traits, the whole genome sequence of FD48 was compared with its related strains. The whole genome analysis revealed a single chromosome with a total length of 3,752,533 bp (3.7 Mb) and an average G + C ratio of 41.19%. A total of 4029 genes were predicted, of which 3964 (98.4%) were protein-encoding genes (PEGs) and 65 (1.6%) were non-protein-coding genes. The interaction of FD48 with the host plants is associated with many chemotactic and motility-related genes. The ability of FD48 to colonize plants and maintain plant growth under adverse environmental conditions was evidenced by the presence of genes for plant nutrient acquisition, phytohormone synthesis, trehalose, choline, and glycine betaine biosynthesis, microbial volatile organic compounds (acetoin synthesis), heat and cold shock chaperones, translation elongation factor TU (Ef-Tu), siderophore production, DEAD/DEAH boxes, and non- ribosomal peptide synthase clusters (bacilysin, fengycin, and bacitracin). This study sheds light on the drought stress-resilient mechanism, metabolic pathways and potential activity, and plant growth-promoting traits of B. altitudinis FD48 at the genetic level.

Comparative genomic analysis provides insight into the phylogeny and potential mechanisms of adaptive evolution of Sphingobacterium sp. CZ-2

Sphingobacterium is a class of Gram-negative, non-fermentative bacilli that have received widespread attention due to their broad ecological distribution and oil degradation ability, but are rarely involved in infections. In this manuscript, a novel Sphingobacterium strain isolated from wildfire-infected tobacco leaves was named Sphingobacterium sp. CZ-2. NGS and TGS sequencing results showed a whole genome of 3.92 Mb with 40.68 mol% GC content and containing 3,462 protein-coding genes, 9 rRNA-coding genes and 50 tRNA-coding genes. Phylogenetic analysis, ANI and dDDH calculations all supported that Sphingobacterium sp. CZ-2 represented a novel species of the genus Sphingobacterium. Analysis of the specific genes of Sphingobacterium sp. CZ-2 by comparative genomics revealed that metal transport proteins encoded by the troD and cusA genes could maintain the balance of heavy metal ion concentrations in the internal environment of bacteria and avoid heavy metal toxicity while meeting the needs of growth and reproduction, and transport proteins encoded by the malG gene could keep nutrients required for the survival of bacteria. Synteny and genome evolutionary analyses of Sphingobacterium strains implicated that the gene family contraction as a major process in genome evolution, with insertional sequences leading to mutations, deletions and reversals of genes that help bacteria to withstand complex environmental changes. Complete genome sequencing and systematic comparative genomic analysis will contribute new insights into the adaptive evolution of this novel species and the genus Sphingobacterium.

Complete genome sequence analysis of a novel alkane-degrading bacterial strain, Acinetobacter vivianii KJ-1, and its diesel degradation ability

With the increasing demand for diesel in various countries, the ecological pollution caused by the improper use, storage, and accidental leakage of diesel needs to be addressed urgently. As an environmentally friendly and cost-effective method, bioremediation generally uses various microorganisms to remove pollutants from the environment. Here, the strain KJ-1, obtained through an enrichment culture using n-dodecane from oil-contaminated soil near a gas station as the substrate, was identified as Acinetobacter vivianii according to its morphology, biochemistry, and molecular biology. The isolate KJ-1 was able to use diesel as a sole carbon source and more than 40% of diesel was degraded after 12 days of incubation with strain KJ-1 in mineral salts medium. The most suitable diesel concentration and nitrogen source concentrations were 4,140 mg/L and 350–700 mg/L, respectively, for diesel degradation and bacterial growth. The optimal initial pH and temperature for strain KJ-1 growth and diesel degradation were 6.5–8.0 and 20–37°C, respectively. To investigate the diesel-degrading mechanisms of this strain, the complete genome was sequenced and annotated. The complete genome consists of one chromosome with a total length of 3,927,757 base pairs and a G + C content of 41.5%. The genes related to the two-component regulatory system and alkane degradation were analyzed. In addition, two putative alkane monooxygenases were analyzed, and the protein sequences were characterized and compared with other AlkBs in Acinetobacter spp. using sequences downloaded from NCBI. The results demonstrated that A. vivianii KJ-1 may be particularly useful for future bioremediation of diesel-polluted soil.

Clinical Epidemiology, Pathology, and Molecular Investigation of Lumpy Skin Disease Outbreaks in Bangladesh during 2020-2021 Indicate the Re-Emergence of an Old African Strain.

Lumpy skin disease (LSD) emerged in Bangladesh in mid-2019, leading to great economic losses for cattle farmers. This study describes the recent occurrence of the LSDV in Bangladesh and examines the clinical manifestation of the disease in local cattle breeds, characteristic epidemiological features, and pathological findings in affected animals. In addition, a full-genome sequencing of two local LSDV isolates was carried out. A total of 565 animals from 88 households were investigated, and 165 samples (skin lesions, saliva, nasal discharge, feces, and milk) were collected for virus detection. Pathology and immunohistochemistry were performed on nodule biopsies. Fever, nodular skin lesions, and swelling of the joints were the most common clinical manifestations. Skin lesions had a higher concentration of viral DNA compared to other sample types and were therefore selected for virus isolation and characterization. Pathology of the LSD skin nodules comprised a granulomatous reaction in the dermis and hypodermis that extended to the surrounding tissues. Development of the skin lesions started with swelling of keratinocytes with cytoplasmic vacuolation, vasculitis, panniculitis, thrombosis, and infarction. Altogether, the LSDV produced transmural, hemorrhagic, necrotizing, proliferative and ulcerative dermatitis. The LSD viral antigen was detected occasionally in the macrophages, epithelial cells, and vascular smooth muscle cells. The complete genome sequence analysis revealed that the two Bangladeshi field strains (BD-V392.1 and BD-V395.1) were distinct from the contemporary field strains and were closely related to the ancestral African Neethling strain. The findings of this study will improve the diagnosis, monitoring, and control of LSD in Bangladesh.