Canine Parvovirus Research Articles

Detection of Canine Parvovirus Type 2 by Designing Multiple Methods and Genetic Characterization in Iran.

Canine parvovirus infection is the most highly infectious in dogs younger than six months. Our study aimed to design and optimize an In-house PCR Assay for Rapid Detection of parvovirus type 2 and compares it with REAL-TIME PCR and LAMP Assay and phylogenetic analysis. The virulence gene selected for the categories was vp2 for CPV-2. PCR products were cloned in pTZ57R/T plasmid for preparation of positive control. Determination of the specificity of primers was done with the negative control virus genomes, and the limit of detection was determined for the Homemade PCR, REAL-TIME PCR, LAMP, and to perform a phylogenetic study using partial vp2 gene sequences. Added analysis of PCR products using agarose gel electrophoresis for the vp2 gene showed 485bp, and GAPDH 900 bp bands, respective amplification using negative control genomes as template was negative. The least detectable copy number for the vp2 gene in a 25 µl PCR reaction equals 19 copies by homemade PCR, LAMP, and REAL-TIME PCR 25 and 21 copies, respectively. The phylogenetic analysis for the five field sequences formed three distinct clusters. The in-house PCR has advantages such as high specificity, sensitivity, and the ability to detect major CPV-2 pathogens. This assay may replace the previous laboratory methods and work as an essential supplement to the more time-consuming assays. Phylogenetic analysis is necessary for epidemiological studies to control and prevent disease.

Characterization of the VP2 and NS1 genes from canine parvovirus type 2 (CPV-2) and feline panleukopenia virus (FPV) in Northern China.

Canine parvovirus type 2 (CPV-2) and feline panleukopenia virus (FPV) cause severe disease in young animals, pups, and kittens. CPV-2 evolved from FPV by altering the species-specific binding of the viral capsid to the host receptor, i.e., the transferrin receptor (TfR), and CPV-2 genetic variants have been identified by specific VP2 amino acid residues (297, 426). Early studies focused on the main capsid protein VP2; however, there have been limited studies on the non-structural protein NS1. In this study, we identified the genetic variants of clinical samples in dogs and cats in northern China during 2019-2020. The genetic characterization and phylogenetic analyses of VP2 and NS1 gene were also conducted. The results revealed that the CPV-2c was identified as the major genetic variant. One new CPV-2b and two CPV-2c strains were collected from cats. Four mutation sites (60, 630, 443, and 545 amino acid residues) were located in the functional domains of the NS1 protein. The phylogenetic analysis of VP2 and NS1 genes showed that they were clustered by geographical regions and genotypes. The gene mutation rate of CPV-2 was increasing in recent years, resulting in a complex pattern of gene evolution in terms of host preference, geographical selection, and new genetic variants. This study emphasizes that continuous molecular epidemiological surveillance is required to understand the genetic diversity of FPV and CPV-2 strains.

Two-Shots of Immunization Effectively Induce Protective IgG against Canine Parvo-Viral Infection

Regardless of good hygiene and management, a significant number of dogs are lost every year due to deadly attack of parvo virus. The immunization schedules recommended by the World Small Animal Veterinary Association (WSAVA) do not have exclusive suggestions on dogs living in Bangladesh. The WSAVA Vaccination Guidelines Group (VGG) suggested that vaccination recommendations that apply to a developed country may not be appropriate for a developing country. Due to inadequate sero-monitoring, emergence and re-emergence of Parvo viral infection following immunization is a common event. Therefore, the current study is designed to establish a definite guideline for Parvo viral immunization in dogs. We immunized group of dogs with commercially available parvo virus vaccine Nobivac Dog®, and a second dose was injected 14-days following the initial dose. Sera samples were collected before starting injections and 7-day following 1st and 2nd dose of vaccinations. We performed Enzyme Linked Immunosorbent Assay (ELISA) to detect protective serum IgG titer against the canine parvo virus type 2 antigen. Demographic data of the participated animals was compared to the status of antibody titer. Findings of this study revealed that two-shots of immunizations with a booster injection of 2-3 weeks apart from the primary immunization is essential to develop positive titer. The purebred female dogs of older than three-months had higher IgG when compared to local, crossbred, male and younger dogs. The results will provide valuable information to establish National Guidelines for Immunizations in Dogs of Bangladesh for effective prevention and control of this deadly disease.

Carnivore protoparvovirus 1 (CPV-2 and FPV) Circulating in Wild Carnivores and in Puppies Illegally Imported into North-Eastern Italy.

The illegal trade of animals poses several health issues to the global community, among which are the underestimated risk for spillover infection and the potential for an epizootic in both wildlife and domestic naïve populations. We herein describe the genetic and antigenic characterization of viruses of the specie Carnivore protoparvovirus 1 detected at high prevalence in puppies illegally introduced in North Eastern Italy and compared them with those circulating in wild carnivores from the same area. We found evidence of a wide diversity of canine parvoviruses (CPV-2) belonging to different antigenic types in illegally imported pups. In wildlife, we found a high circulation of feline parvovirus (FPV) in golden jackals and badgers, whereas CPV-2 was observed in one wolf only. Although supporting a possible spillover event, the low representation of wolf samples in the present study prevented us from inferring the origin, prevalence and viral diversity of the viruses circulating in this species. Therefore, we suggest performing more thorough investigations before excluding endemic CPV-2 circulation in this species.

Competitive ELISA test system for the detection of antibodies to the rabies virus in animals (Rhabdoviridae: Lyssavirus)

The main approach to the rabies prevention is the vaccination of domestic and wild carnivores. For the routine evaluation the anti-rabies vaccination effectiveness, World Organization for Animal Health (OIE) recommends various enzyme-linked immunosorbent assays (ELISA).The aim of the study was to design and validate a competitive ELISA (cELISA) test system for the detection of antibodies to the rabies virus (RABV). The development of the cELISA was carried out following the OIE recommendations. The repeatability of the cELISA results within one laboratory was satisfactory (coefficient of variation 7.95-13.61%). The coefficient of determination (CD) between the results of the virus neutralization reaction (FAVN) and cELISA was 0.988, p < 0.001. The lower threshold for antibody detection was less than 0.02 IU/ml. The cELISA did not demonstrate cross-reactivity against antibodies to canine distemper virus, parainfluenza virus, parvovirus, coronavirus, and canine adenovirus (types I and II). During the study of 137 dog blood sera, diagnostic specificity (DSp) and diagnostic sensitivity (DSe) for the cELISA were 83.1% and 94.9%, respectively, and CD between the cELISA and FAVN results was 0.968, p < 0.001. Indirect ELISA test systems for determining the level of antibodies to RABV are not sensitive enough compared to reference tests, unlike cELISA. The developed test system is not inferior for its DSp and DSe to the commercial cELISA BioPro ELISA Rabies Ab (DSp 66.7%, DSe 94.4%). The developed cELISA test system can be used to detect antibodies to RABV in the blood serum of dogs for evaluating the effectiveness of mass vaccination programs.

Identification of novel recombinants and proposed standard reference genomes for phylogenetic classification of canine parvovirus-2 (CPV-2): Comprehensive analysis revealing global evolutionary trait.

Sustained spread and continuous evolution of CPV-2 generate new genetic information; nevertheless, there is no adopted phylogenetic tool, and parvo virologists still refer to the three antigenic variants. Herein, this report attempted to review the evolutionary trait of CPV-2 and proposed standard reference genomes using the Maximum Likelihood-based phylogenetic analysis and Parsimony-Informative Sites. The analysis revealed three main evolutionary pathways where CPV-2 strains cluster into distinct clades depicted as GI, GII, or GIII, respectively. Furthermore, novel CPV-2 natural recombinants were detected, occurring only between the newly identified strains (2017-2020). Those findings provide unique insights into the evolutionary relatedness of CPV-2, clarify discrepancies between different geographic areas and will contribute to achieving a more reliable CPV-2 genetic and evolutionary genotyping classification.

Prevalence and Characteristics of Canine Parvovirus Type 2 in Henan Province, China.

To investigate the epidemic profile and genetic diversity of canine parvovirus type 2 (CPV-2), a total of 111 clinical samples collected from dogs suspected of CPV-2 infection in 10 cities of Henan province of China during 2020 to 2021 were screened by PCR. The results showed a CPV-2-positive rate of 88.29% (98/111). Nearly full-length genomes of 98 CPV-2 strains were sequenced and analyzed. CPV-2c strains (91.84%, 90/98) were significantly higher than that of new CPV-2a strains (8.16%, 8/98) in Henan province without detecting other CPV genotypes, indicating that CPV-2c has become the dominant genotype in Henan province. A phylogenetic analysis of NS1 and VP2 amino acids grouped the strains in this study with Asian strains, which clustered into an identical branch. Based on the CPV-2 VP2 sequences in this study and available in the NCBI database, the adaptation analyses showed that 17 positive selection sites and 10 parallel evolution sites were identified in the VP2 protein of CPV-2, of which three sites (sites 5, 370, and 426) were both under positive selection pressure and parallel evolution. Interestingly, two amino acid mutations (A5G and Q370R) were also observed in the VP2 proteins of 82 CPV-2c strains in this study, which differed from the earlier CPV-2c strain (GU380303) in China. In addition, a unique mutation (I447M) was observed in the VP2 protein of five CPV-2c strains, which was first reported in China. This study provides powerful insight to further our understanding of the epidemic status and evolution of CPV-2 in China. IMPORTANCE CPV-2 was the original virus strain identified in dogs, which cause an acute and lethal disease in dogs. Subsequently, the original CPV-2 was replaced throughout the world by novel antigenic variants (e.g., CPV-2a, CPV-2b, new CPV-2a, new CPV-2b, and CPV-2c). Currently, the epidemiological characteristics of CPV-2 in Henan province of China is still unclear. In our study, a total of 98 nearly full-length genomes of CPV-2 strains were obtained to explore prevalence and genetic evolution of CPV-2 in Henan Province. Moreover, the epidemiological and genetic evolution of CPV-2 in China since its discovery was also investigated. The results of this study will provide valuable information regarding the evolution of CPV-2 strains in China.

Persistence of DNA from canine parvovirus modified-live virus in canine tissues.

Canine parvovirus (CPV-2) modified-live virus vaccine strain can replicate in lymphoid tissues and intestinal mucosa after administration, being shed through canine faeces. Detection of vaccine strains has been reported in the bloodstream and faeces, potentially interfering with molecular diagnostic tests. The persistence of these strains in canine tissues has not yet been described. With this aim, canine tissues were tested during a molecular survey to screen for the presence of canine enteric viruses. Tissue samples from 165 dead dogs were tested by a conventional PCR assay. Positive samples and five commercial vaccines were subjected to sequence analysis. Vaccinal strains were detected and virus load was measured by using a set of real-time PCR assays using minor-groove binder (MGB) probes. Seventy-five dogs (45.4%) tested positive for CPV-2. Strains from 70 dogs were characterised as field variants. The presence of CPV sequences of vaccine origin was observed in the spleen, intestine, and mesenteric lymph nodes of five young dogs. Vaccinal strains were detected from 12 to 24 days after the last vaccine administration. Viral loads comprised between 6.3 × 102 and 9.95 × 104 DNA copies/10 µl of template. This study confirms that CPV vaccinal strains can be detected in canine tissues after vaccination, so post-mortem diagnosis of CPV infection needs further molecular analyses to assess the viral type (vaccine or field strains). The present study updates the current information on the persistence of CPV vaccine strains in canine tissues and their possible interference with molecular assays.

Detection of Canine Viral and Bacterial Agents Associated with Gastroenteritis by PCR and RT-PCR

Background: The important causative agents associated with gastroenteritis include Canine Parvovirus, Canine Rotavirus, Canine coronavirus and E. coli. The diagnostic assay viz. lateral flow test, PCR, RT-PCR was used for screening of diarrheal fecal samples of dogs to detect the presence of these pathogens. Methods: The present study envisaged the appraisal of Canine parvovirus, Rotavirus, corona virus and enterotoxigenic E. coli infection. Total 50 rectal swabs/fecal swab in duplicate were collected for detection of viral andbacterial enteropathogens associated with canine gastroenteritis. For virus detection by PCR a 10% fecal suspension was prepared in phosphate buffer saline (PBS) (pH 7.2). The detection of viral and bacterial agents done by as per reference methods. Result: Out of 50 fecal smples 10(20%) samples were found positive, 2(4%) were found to be positive for Canine Rotavirus and None of sample was found positive for Canine Corona virus. Canine Parvo virus 15(30%) and 1(2%) for Canine rotavirus by PCR and RT-PCR and None of the sample was positive for Canine Corona virus. Of this Canine parvovirus, Rotavirus and E. coli were recovered either singly or in association with each other in the form of mixed infection. Out of 50 samples, 15(30%) and 1(2%) were found positive for Canine parvovirus, Rotavirus and 45(90%) E. coli.

First genetic detection and characterization of canine parvovirus type 2 (Carnivore protoparvovirus 1) in southwestern Ethiopia

The species Carnivore protoparvovirus 1 includes viruses, e.g. canine parvovirus (CPV-2) and feline panleukopenia virus (FPV), which are among the most relevant for pets, causing extremely severe clinical signs and high fatality rate in dogs and cats. Moreover, a broad range of wild hosts, including endangered ones, were proven to be susceptible. Currently, no data on CPV-2 molecular epidemiology and strain characterization are available in Ethiopia, also considering the frequent contacts between domestic and wild populations. In the present study, a molecular epidemiology survey was performed on 92 fecal samples collected from domestic (n = 84) and stray (n = 8) dogs in southwestern Ethiopia in 2021. Approximately, 10% of the samples tested positive and the complete VP2 sequences of 5 strains were obtained, classified within the CPV-2a (n = 1) and CPV-2c (n = 4) antigenic variants. In most instances, the closest genetic relatives were strains of Chinese origin, which is fully compatible with the intense relationships that have been developing between the two countries, involving human being travels and their pets as well. Considering the clinical relevance of this infection and the risk it poses to local domestic and wild carnivore populations, especially endangered ones, much stronger monitoring and surveillance activity on foreign incoming animals should be performed. More strict constraints on animal introduction, e.g. compulsory vaccination, should also be considered.

Epidemiology of canine parvovirus infection in and around Pantnagar, Uttarakhand: A retrospective study

In India, Canine Parvovirus Infection is an endemic viral disease-causing severe gastroenteritis and significant numbers of deaths in puppies, even in vaccinated populations. A retrospective study was conducted between June, 2021 and June, 2022 in and around Pantnagar, Uttarakhand, in which cases of gastro-enteritis were screened for canine parvovirus infection. A total of 258 cases out of 627 cases presented for gastro-enteritis were found to be positive for canine parvovirus based on Rapid Antigen Tests and Polymerase Chain Reaction with a prevalence rate of 41.15%. Data associated with factors such as age, breed, sex, season, immunisation and relocation stress were recorded. Mongrels were found to be the most affected among various breeds, with a prevalence rate of 51.16%, followed by the exotic breed Labrador retriever (9.68%). Males (63.57%) were more found to be affected more than females (34.43%). As for age, prevalence was higher in the age group of 3-6 months (43.40%), followed by less than 3 months of age (31.40%) respectively. Considering other risk factors such as season, vaccination status and relocation stress, prevalence was seen to be higher in to be higher comparatively in spring (33.33%) and winter (29.07%); also, higher prevalence in non-vaccinated (63.13%) and about 25.19% of the animals which were relocated recently were found to be infected with canine parvovirus.

Seroepidemiological and Clinicopathological Investigation of Canine Coronavirus Infection in Dogs, in Türkiye

Domestic and wild dogs of all ages and breeds are susceptible to Canine Coronavirus (CCoV) infections and be seen in Türkiye and amongst world. CCoV has recently been declared a zoonotic disease agent and the eighth pathogenic human coronavirus. This study was conducted on 143 naturally infected dogs with gastroenteritis which were not vaccinated against CCoV in Türkiye in 2015-2020. The data of dogs were analyzed seroepidemiologically, clinicopathologically and statistically. CCOV antibodies in serum and CCOV antigens in stool were detected by ELISA and lateral immunochromatography. The rising CCoV IgG antibody titers were detected at all dogs and were as follows; 64 ng/L in 81 (81%) dogs. CCOV and Canine Parvovirus (CPV) antigen were detected together in the stool of the 41 (28.7%) dogs. As a result, it was concluded that the CCOV agent is in circulation among dogs living in Türkiye. CCOV and CPV can cause co-infections and increased mortality. Although infection can be seen in dogs of all ages, it can be seen more frequently in dogs younger than 1 year of age, and especially in dogs younger than 6 months, and can cause enteritis, low hemoglobin, erythropenia, lymphopenia, leukopenia, thrombocytopenia, and hypoproteinemia.

Emergence of canine parvovirus type 2c (CPV-2c) of Asian origin in domestic dogs in Myanmar

Canine parvovirus type 2 (CPV-2) causes severe gastroenteric disease in domestic dogs and wild canids. This study aimed to conduct molecular detection and characterization of CPV-2 in domestic dogs in Myanmar from December 2017 to October 2019. Rectal swabs (n = 143) were collected from domestic dogs from shelters and small animal hospitals in Yangon, Myanmar. CPV-2 detection was performed by a PCR assay targeting the VP2 gene. Our result showed that 25.17% (36/143) of swab samples tested positive for CPV-2. CPV-2 strains (n = 15) were selected for complete VP2 gene sequencing. Phylogenetic analysis showed that CPV-2 strains from Myanmar clustered together with Asian CPV-2c from China, Indonesia, Taiwan and Thailand but in separate clusters from CPV-2c from Europe and North America. Characteristic amino acid at residues 267Y and 324I were observed in CPV-2c strains from Myanmar, suggesting the Asian origin. In conclusion, our findings expanded the evidence of the predominance of CPV-2c in Southeast Asia. Thus, the surveillance of CPV-2 in domestic dogs in the countries and regions should be routinely conducted to provide epidemiological information for supporting prevention and control practices.

Molecular epidemiology of canine parvovirus in Namibia: Introduction pathways and local persistence

Canine parvovirus is a member of the Carnivore protoparvovirus 1 species that, after a relatively recent origin, has reached a worldwide distribution. Like other ssDNA viruses, it is featured by a remarkable evolutionary rate and thus genetic variability. CPV-2 is responsible for a severe systemic infection affecting especially domestic dogs. However, other carnivores, including wild species, are susceptible and thus represents a menace to wildlife conservation too. Despite the relevance of the topic, molecular epidemiology data are scarce and outdated in certain areas of the world, like Africa and, in particular, Namibia. The present study investigates the occurrence and genetic features of CPV in Namibian domestic dogs and jackals. The VP2 of detected strains was characterized and analyzed to assess the viral circulation and link among host species, Namibian districts and foreign countries. With the only exception of one New-CPV-2a, all the detected strains belonged to the CPV-2c antigenic variant and were closely related to strains of Asian origin. Nevertheless, a dedicated phylogeographic analysis revealed that the introduction was more likely mediated by other African countries, highlighting the challenge of controlling illegal animal imports across land borders. Similarly, the absence of any geographical clustering within Namibia testify a substantially unconstrained viral circulation among districts. The absence/incomplete vaccination status reported by the animal owners could have significantly contributed to the infection's success after its introduction. Finally, infection of a wild jackal was also proven. Although the limited wild animals' sample size prevents any definitive conclusion, the identity of the sequences from the jackal and the ones originating from the domestic dogs suggests a potential inter-species transmission. The epidemiological and clinical implications in wild specie remain obscure.

Clinical complications for canine parvovirus infection in puppies

Canine parvovirus (CPV) infection is one of the most important infectious diseases affecting young puppies. With aggressive supportive treatment, most puppies tend to survive the infection, while a minority develops some complications that make the course of treatment longer and more complicated. In this study, we documented the main complications of the CPV infection in 44 puppies referred to private practices in Baghdad. We present a full description of the cases with their clinical, laboratory and imaging findings and discuss the type, rate and causes of the complications in the survived puppies that occurred in the next two weeks. Out of 44 puppies, five did not survive, while, 30 cases were treated without any complications. In the meantime, Carpal / Tarsal laxity, decreased growth rate, antibiotic responsive diarrhea and ascites were observed in 6, 2, 2 and 1 case, respectively. Weaker puppies, inappropriate treatment protocol and/or uncomfortable small enclosures could be important predisposing factors for these complications. Therefore, it is highly recommended to start an appropriate treatment in larger enclosures and more comfortable floor as soon as possible, in addition to providing the oral soft food no later than 3-5 days to reduce further complications.

Isolation and identification of a novel canine parvovirus type 2c strain in domestic cats in Dalian, China.

Canine parvovirus (CPV) and feline panleukopenia virus (FPV) are highly contagious and cause severe enteric diseases, with high mortality rates in dogs and cats. In the present study, we isolated and identified a novel CPV-2c strain (FPV-DL04 strain) from 18 cats with gastroenteritis symptoms and a positive parvovirus PCR test result in Dalian, China. Molecular characterization, sequence analysis, and phylogeny determination were performed on the VP2 gene of this strain. The results showed that the FPV-DL04 strain had 99.4% homology with the CPV-2c CN/HN1708 strain, and both strains had S297A and A300G key mutation sites. Interestingly, we also found that the DL04 strain has a A5G mutation site, but no F267Y and Y324I mutation sites. This study provided new important findings regarding the evolution of parvovirus infection in domestic cats in China.

Evaluation of a simple ultrafiltration method for concentration of infective canine parvovirus and feline coronavirus from cell culture supernatants

Enrichment of viral infectious titers following its propagation by cell culture is desirable for various experimental studies. The performance of an ultrafiltration (UF) process to concentrate infectious titers of non-enveloped Canine parvovirus 2 (CPV-2) and enveloped Feline coronavirus (FCoV) obtained from cell culture supernatants was evaluated in this study, and compared with ultracentrifugation (UC) process. A mean gain of > 1.0 log10 TCID50/mL was obtained for CPV-2 with UF, which was comparable with the gain obtained by UC. On the other hand, the gain was lower (0.7–1.0 log10 TCID50/mL) for FCoV with UF in contrast to UC (> 2.0 log10 TCID50/mL). However, the lower retentate volume following UC (∼120 fold) compared to that following UF (∼10 fold) for either of the viruses suggests a trend of increased infectious titer retention in UF concentrates relative to UC concentrates. The simplistic UF process evaluated here thus has the potential for use in applications requiring increased infectious titers of CPV-2 and FCoV.

The Changes in Canine Parvovirus Variants over the Years.

Canine parvovirus (CPV-2) is one of the most important pathogens in dogs, and despite the continual development of vaccines against CPV-2, CPV-2 is still circulating in the canine population. The CPV-2a/2b/2c variant has replaced the original CPV-2 virus and seems to exhibit accelerated transmission. Although CPV-2 infection has been frequently reported, no studies have summarized information of CPV-2 variants currently circulating worldwide. To track the evolution of CPV-2, we downloaded and analyzed all VP2 sequences from the NCBI database (from 1978 to 2022). We found that CPV-2c shows a tendency to replace CPV-2a as the new dominant variant in Asia, South America, North America and Africa. Additionally, CPV-2c, which is prevalent in most regions of Asia, carries two special mutations in VP2, A5G and Q370R, and has become a dominant mutation with spillover already occurring. In conclusion, this summary of the types of global epidemic variants provides new insight into the evolution of CPV-2 and raises awareness for blocking the spread of this virus. The spread of Asian-derived CPV-2c urgently needs to be further under surveillance.

Antiviral activity of canine interferon lambda 3 expressed using a recombinant adenovirus against canine coronavirus, canine parvovirus, and canine distemper virus.

Canine coronavirus (CCoV), canine parvovirus (CPV), and canine distemper virus (CDV) are highly contagious canine pathogens; dogs with these diseases are difficult to treat. In a previous study, we developed a recombinant adenovirus expressing canine interferon lambda 3 (Ad-caIFNλ3) in canine epithelial cells. In this study, we aimed to investigate the antiviral activity of Ad-caIFNλ3 against CCoV, CPV, and CDV in two canine cell lines, A72 and MDCK. Ad-caIFNλ3 transduction suppressed replication of these viruses without cytotoxicity. Our results suggest that Ad-caIFNλ3 may be a therapeutic candidate for canine viral diseases.Supplementary InformationThe online version contains supplementary material available at 10.1007/s11259-022-10000-1.

Identification and Molecular Characterization of a Divergent Asian-like Canine Parvovirus Type 2b (CPV-2b) Strain in Southern Italy.

Canine parvovirus type 2 (CPV-2) is an infectious agent relevant to domestic and wild carnivorans. Recent studies documented the introduction and spread of CPV-2c strains of Asian origin in the Italian canine population. We investigated tissue samples from a puppy collected during necropsy for the presence of viral enteropathogens and all samples tested positive only for CPV-2. The full coding sequence of a CPV-2b (VP2 426Asp) strain was obtained. This virus was related to CPV-2c strains of Asian origin and unrelated to European CPV-2b strains. The sequence had genetic signatures typical of Asian strains (NS1: 60Val, 545Val, 630Pro; VP2: 5Gly, 267Tyr, 324Ile) and mutations rarely reported in Asian CPV-2b strains (NS1: 588N; VP2: 370Arg). Phylogenetic analyses placed this strain in well-supported clades, including Asian CPV-2c-like strains, but always as a basal, single-sequence long branch. No recombination was observed for this strain, and we speculate that it could have originated from an Asian CPV-2c-like strain that acquired the 426Asp mutation. This study reports the first evidence of an Asian-like CPV-2b strain in Italy, confirming the occurrence of continuous changes in the global CPV-2 spread. Since positive convergent mutations complicate data interpretation, a combination of phylogenetic and mutation pattern analyses is crucial in studying the origin and evolution of CPV-2 strains.