CPV-2a Strains Research Articles

Molecular characterization and genetic diversity of parvoviruses prevalent in cats in Central and Eastern China from 2018 to 2022.

Cats are a potential source of genetic diversity for parvoviruses. Herein, 134 samples were collected from cats with clinical gastroenteritis and analyzed for the presence of viral DNA via polymerase chain reaction, which revealed 48 positive samples. Identity analysis of VP2 nucleotide sequences indicated that these 48 strains, belonging to feline panleukopenia virus (FPV) and canine parvovirus type-2 (CPV-2; including new CPV-2a and CPV-2c genotypes), shared 94.59-99.94% nucleotide identity with the reference strains. The FPV strain F8 (isolated from Vietnam) appeared to be a recombinant of strains HB2003 and JS1901, whereas the Chinese CPV-2b strain BM-(11) isolated in 2011 was believed to be a recombinant of strains AH2008 and JS1901. In phylogenetic tree analysis based on VP2 nucleotide sequences, all obtained FPV strains and most reference FPV strains were clustered together, except strain BJ-22, which originated from monkeys. Further, two new CPV-2a strains (AH2005 and AH2008) were close to the newly reported Chinese CPV-2a strains but were distant from the other CPV-2a strains, namely CPV-339 (from the United States) and K022 (from South Korea). Additionally, the FPV and CPV-2 strains had high mutation rates in the antigenic regions of the VP2 protein. According to model prediction of the CPV-VP2 protein, these mutations may cause changes in the tertiary structure of VP2. The findings of this study can be used to improve the pre-evaluation of vaccination efficacy against diseases caused by FPV and CPV-2 in domestic cats and understand their genotypic transmission and mutation trends.

Genomic Sequence and Phylogenetic analysis of Canine Parvovirus New 2a Variant Strain in Indian Dogs

In this study, a virulent field canine parvovirus type - 2a variant (CPV-new 2a) strain, ABT/CPV/MVC02, was identified from the faeces of a Dog with severe haemorrhagic gastroenteritis in Chennai, Tamil Nadu, India. The genomic sequence of ABT/CPV/MVC02 (MN661243) shares 99.33% nucleotide identity with those of CPV new 2a variant strains. Based on the VP2 capsid protein amino acid variation (Ser297Ala), the ABT/CPV/MVC02 strain is classified as a new CPV 2a variant strain. Two VP2 mutations Tyr324Ile and Thr440Ala were reported in most of the CPV-2a stains also present in ABT/CPV/MVC02.The phylogenetic analysis of the reported strain revealed that it was distinct from existing CPV 2a strains from India and other countries. This complete genomic analysis revealed that the CPV-2a new variant strain circulating in India is evolving rapidly with unique antigenic variations between field CPV-2a and vaccine strains which may be the major cause for vaccine failure in vaccinated dogs.

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.

Fatal canine parvovirus type 2a and 2c infections in wild Chinese pangolins (Manis pentadactyla) in southern China.

The Chinese pangolin (Manis pentadactyla) is a critically endangered scale-covered mammal belonging to the order Pholidota. Wild pangolins are notably susceptible to pathogen infection and are typically characterized by impoverished health. However, little is currently known regarding the viruses prevalent among pangolins. In this study, we report the detection of two subtypes of canine parvovirus type 2 (CPV-2), namely CPV-2a and CPV-2c, both of which caused severe diarrheal disease in two post-rescue pangolins with fatal consequences. As in CPV-2-infected dogs, intensive lesion of the mucosal layer of the small intestines is a prominent feature in infected pangolins. Moreover, the immunochemistry results demonstrated that CPV-2 antigens were distributed in the crypts of small intestine. Additionally, typical parvovirus-associated CPV-2 were detected after four passages in F81 cells, and typical parvovirus-like particles, approximately 20nm in diameter, were observed in the cell supernatants. Phylogenetic analysis revealed that the VP2 viral protein sequences (GenBank accession number OP208805) isolated from one pangolin (termed P1) were classified as CPV-2c, with 99.8% identity to a CPV-2c strain (MN832850) isolated from a Taiwanese pangolin found in Taiwan Province. In contrast, VP2 sequences (#OP208806) obtained from the second pangolin (P2) were classified as CPV-2a, with 99.8% identity to a CPV-2a strain (KY386858) isolated from southern China. In this study, we thus confirmed the infection of pangolins with CPV-2c in mainland China and demonstrated that CPV-2a also can infect pangolins. Based on these findings, we recommend that further investigations should be conducted to establish the interspecies transmission of these viruses among wild pangolins, wild carnivores, and stray dogs.

Distribution and diversity of dog parvoviruses in wild, free-roaming and domestic canids of Newfoundland and Labrador, Canada.

Some parvoviruses of carnivorans can infect multiple host species. Since many canine parvoviruses were only discovered recently, their host-range is still unexplored. We examined the host distribution and diversity of five dog parvoviruses in four canine populations from Newfoundland and Labrador, Canada, and investigated the potential for these viruses to cross the species barriers. Canine bocavirus 2 (CBoV-2) and the minute virus of canines were detected in stool from free-roaming dogs from Labrador (5/48 [10.4%] and 3/48 [6.3%], respectively) and two different CBoV-2 variants were identified. Canine bufavirus was identified in stool from free-roaming dogs (1/48, 2.1%) and foxes (3/80, 3.8%) from Labrador, but two different variants were observed in the two host species. The variant found in foxes was highly divergent from previously identified strains. Two cachavirus 1 variants, genetically similar to those circulating in other Canadian wildlife, were found in spleens from Newfoundland coyotes (3/87, 3.5%). Canine parvovirus type 2 (CPV-2) was found in stool from free-roaming dogs from Labrador (2/48, 4.2%) and in spleens from Newfoundland coyotes (3/87, 3.5%). Comparing CPV-2 sequences from these hosts to those retrieved from local symptomatic domestic dogs revealed the presence of a highly heterogeneous viral population as detected strains belonged to five different clades. The close relationship between CPV-2a strains from a dog and a coyote suggests the occurrence of viral transfer between wild and domestic canids. The identification of highly related strains with a similar molecular signature characteristic of older CPV-2 strains in free-roaming and domestic dogs suggests a probable common ancestry and that older CPV-2 strains, which have not been identified in dogs since the 1990s, persist in this part of Canada. Follow-up studies should evaluate samples from a larger number of animals and host species to extensively investigate the possible occurrence of cross-species transmission for recently discovered parvoviruses.

Molecular characterization of canine parvovirus from domestic dogs in Nigeria: Introduction and spread of a CPV-2c mutant and replacement of older CPV-2a by the "new CPV-2a" strain.

Canine parvovirus (CPV) is a contagious and highly pathogenic virus of dogs. After its first report in 1978, the CPV original type (CPV-2) was rapidly and totally replaced by three antigenic variants named CPV-2a, CPV-2b and CPV-2c that circulate in various countries at different frequencies and recently reported in Nigeria. This study describes the molecular characterization of 28 CPV strains in dogs presenting with gastroenteritis in veterinary clinics at Lagos and Ibadan, Nigeria. The results show the predominance (92.8%) of CPV-2a, while CPV-2c was found only in two samples. Phylogenetic analyses revealed that the CPV Nigerian strains were closely related to Asian strains and 26 CPV-2a out of 28 CPV sequences fell into 2 different subclades consistent with predicted amino acid mutations at position 267, 321, 324 and 440. Lys321Asn was evident in all the Nigerian strains whilst Phe267Tyr and Tyr324Ile were observed in 96.4% of the sequences, respectively. Thr440Ala occurred in 89.3% of sequences from this study. The new CPV-2a was predominant and appears to have replaced other CPV-2a strains in South-western Nigeria whilst the CPV-2c strain which is identical to the isolate recently reported in Northern Nigeria, may have been introduced in this country at the time of this study. Monitoring virus epidemiology is important to better understand the dynamics of CPV evolution and the eventual need to change or improve existing vaccination strategies.

First report of canine parvovirus molecular detection in Bangladesh.

Background and Aim:Canine parvovirus (CPV) is the most important cause of mortality in dogs in many parts of the world. Clinical cases exhibit characteristic signs, including foul-smelling bloody diarrhea, vomiting, fever, and dehydration. This study assessed field and vaccine variants of parvovirus in the Chattogram metropolitan area, Bangladesh. The investigation also aimed to identify risk factors for this disease. This research is the first to identify the presence of CPV in Bangladesh through molecular examination.Materials and Methods:From October to December 2019, a total of 100 dogs were included in the study. Rectal swabs were taken from all dogs. Twenty dogs showed clinical signs of parvovirus. All clinically affected animals along with 20 randomly selected healthy dogs were tested using amplification refractory mutation system (ARMS)-polymerase chain reaction (PCR) to identify variants from the samples. Logistic regression model analysis was performed to determine the possible risk factors for CPV.Results:ARMS-PCR showed the presence of all three variants, CPV2a, CPV2b, and CPV2c, in clinically ill dogs, and vaccines available in the study area showed either CPV2a or CPV2b strain. The CPV2c variants showed a higher incidence than the other variants. All apparently healthy animals tested were molecularly negative. Multivariable logistic regression model (generalized linear mixed model) indicated that exotic breeds were 3.83 times more likely to be infected by CPV than local breeds. Furthermore, dogs reared in semi-intensive and extensive management systems were 3.64 and 3.79 times more likely to be infected, respectively, than those reared in an intensive management system.Conclusion:These findings provide practitioners and pet owners information on the occurrence of different variants and help design effective prevention strategies for CPV infection.

Molecular epidemiology and genetic evolution of canine parvovirus in East China, during 2018-2020.

Canine parvovirus type 2 (CPV-2) emerged in the late 1970s, which caused high rates of morbidity and mortality in dogs. In last decade, five genetic variants (CPV-2a, CPV-2b, CPV-2c, New CPV-2a, and New CPV-2b) were frequently reported in the dog population, and replaced the original CPV-2, rising widespread concerns. However, little is known about their recent genetic diversity and evolution. The aim of this study was to analyze the characteristics of the CPV-2 strains collected in East China from 2018 to 2020. The 57 CPV-2 strains were isolated from rectal swab samples (n=140). They belong to three different genotypes, based on VP2 protein amino acid sequence. The results revealed a high prevalence of CPV-2c (77.19%) compared to the New CPV-2a (5.26%) and New CPV-2b (17.54%) strains. Further analysis showed that nucleotide homology of the VP2 gene among the 57 CPV strains was 98.9%~100%, and the homology with 24 reference strains from different countries and regions was 98.1%~100%. The phylogenetic tree of VP2 gene sequence showed that 44 CPV-2c strains were distantly related to CPV-2, CPV-2a, CPV-2b, New CPV-2a, New CPV-2b and European/American CPV-2c strains, and were closely related to Asian CPV-2c strains. The results showed that these Asian CPV-2c strains had become the dominant strain, which renewed the knowledge of CPV-2 molecular epidemiology in East China.

Research on factors that influence the course of canine parvovirosis

The aim of this study was to demonstrate correlations of the course of canine parvovirus infection and the rate of cytopathic effect (CPE) on CPV-infected CCC clone 81 cell lines with the age of the affected animals, their history of CPV vaccination and the strain of the virus causing the infection. Forty-five dogs with parvovirus confirmed by rapid diagnostic tests and PCR were studied. Faecal samples from diseased animals were used to infect cell lines, which were then observed for a cytopathic effect. The statistical analysis of the results obtained demonstrated a positive correlation between the recovery rate of the diseased animals and the fact that they had been vaccinated against CPV. In dogs that had been vaccinated against CPV, the mean duration of the disease was 6 days, whereas in the group of unvaccinated dogs it was 7 days. The course of the disease also depended on the virus strain. It was shown that dogs from which the CPV-2a strain was isolated took longer to recover, had more severe clinical signs and higher mortality. The cytopathic effect appeared in all infected cultures, while the rate at which changes occurred was independent of the virus strain with which the cells were infected, the age of the dogs from which CPV was isolated and their CPV vaccination history. The study results obtained may serve as valuable prognostic guidelines as to the duration of and recovery from canine parvovirus infection.

The Efficacy of PIND-ORF with Canine ParvoVirus vaccines in the protection of experimentally challenged puppies against the newly identified CPV-2a virus

Canine parvovirus infection (CPV) is one among several hazard diseases that incorporates a dramatic end and remains a common and vital reason of morbidity and mortality in puppies. The aim of this study is to evaluate the efficacy of local and imported CPV vaccines along with PIND-ORF against the experimentally challenged puppies with the newly identified CPV-2a virus. Forty native breed puppies around 45 days old free from internal and external parasites (as examined clinically) and negative for CPV antibodies (as screened by serum neutralization test) were enrolled, and randomized into five groups (8 animals, each). Vaccinated groups were compared to each other, or to paramunity inducer inoculated group and to neither treatment nor vaccinated group. It was found that both local canine parvovirus and Vanguard are safe and potent vaccines inducing no clinical post-vaccination reaction and high levels of specific CPV antibodies (256) by 30 days post-vaccination. Such vaccines provided 80% protection for challenging puppies against the recent virulent strain (CPV-2a), while, unvaccinated puppies did not withstand the challenging virus infection. Besides, it was noticed that PIND-ORF enhanced the puppies immune response through the 1st 2 weeks post-vaccination, however, it was unable to enhance their ability to guard against the new CPV-2a virus infection strain. So it may be concluded that the currently used CPV vaccines; either the local or imported ones can protect puppies towards new CPV-2a strain along with paramunity inducer PIND-ORF.

Carnivore Parvovirus Ecology in the Serengeti Ecosystem: Vaccine Strains Circulating and New Host Species Identified.

Carnivore parvoviruses infect wild and domestic carnivores, and cross-species transmission is believed to occur. However, viral dynamics are not well understood, nor are the consequences for wild carnivore populations of the introduction of new strains into wild ecosystems. To clarify the ecology of these viruses in a multihost system such as the Serengeti ecosystem and identify potential threats for wildlife conservation, we analyzed, through real-time PCR, 152 samples belonging to 14 wild carnivore species and 62 samples from healthy domestic dogs. We detected parvovirus DNA in several wildlife tissues. Of the wild carnivore and domestic dog samples tested, 13% and 43%, respectively, were positive for carnivore parvovirus infection, but little evidence of transmission between the wild and domestic carnivores was detected. Instead, we describe two different epidemiological scenarios with separate routes of transmission: first, an endemic feline parvovirus (FPV) route of transmission maintained by wild carnivores inside the Serengeti National Park (SNP) and, second, a canine parvovirus (CPV) route of transmission among domestic dogs living around the periphery of the SNP. Twelve FPV sequences were characterized; new host-virus associations involving wild dogs, jackals, and hyenas were discovered; and our results suggest that mutations in the fragment of the vp2 gene were not required for infection of different carnivore species. In domestic dogs, 6 sequences belonged to the CPV-2a strain, while 11 belonged to the CPV-2 vaccine-derived strain. This is the first description of a vaccine-derived parvovirus strain being transmitted naturally.IMPORTANCE Carnivore parvoviruses are widespread among wild and domestic carnivores, which are vulnerable to severe disease under certain circumstances. This study furthers the understanding of carnivore parvovirus epidemiology, suggesting that feline parvoviruses are endemic in wild carnivores in the Serengeti National Park (SNP), with new host species identified, and that canine parvoviruses are present in the dog population living around the SNP. Little evidence of transmission of canine parvoviruses into wild carnivore species was found; however, the detection of vaccine-derived virus (described here for the first time to be circulating naturally in domestic dogs) highlights the importance of performing epidemiological research in the region.

Isolation and sequence analysis of the complete VP2 gene of canine parvovirus from Chinese domestic pets and determination of the pathogenesis of these circulating strains in beagles

Canine parvovirus (CPV) causes acute gastroenteritis in domestic dogs, cats, and several wild carnivore species. In this study, the full-length VP2 gene of 36 CPV isolates from dogs and cats infected between 2016 and 2017 in Beijing was sequenced and analyzed. The results showed that, in dogs, the new CPV-2a strain was the predominant variant (n = 18; 50%), followed by the new CPV-2b (n = 6; 16.7%) and CPV-2c (n = 3; 8.3%) strains, whereas, among cats, the predominant strain was still CPV-2 (n = 9; 25%). One new CPV-2a strain, 20170320-BJ-11, and two CPV-2c strains, 20160810-BJ-81 and 20170322-BJ-26, were isolated and used to perform experimental infections. Multiple organs of beagles that died tested PCR positive for CPV, and characteristic histopathological lesions were observed in organs, including the liver, spleen, lungs, kidneys, small intestines, and lymph nodes. Experimental infections showed that the isolates from the epidemic caused high morbidity in beagles, indicating their virulence in animals and suggesting the need to further monitor evolution of CPV in China.

Application of Duplex Fluorescence Melting Curve Analysis (FMCA) to Identify Canine Parvovirus Type 2 Variants.

Canine parvovirus (CPV-2) is an enteric virus causing morbidity and mortality in dogs worldwide. Since CPV-2 emerged as canine pathogen, the original CPV-2 strain has constantly evolved, and its primary variants (CPV-2a, CPV-2b, and CPV-2c) co-circulate to varying extents in canine populations worldwide. Thus, rapid and accurate laboratory diagnoses of CPV-2 variants are crucial to monitor CPV-2 evolution. Conventional methods for CPV-2 genotyping are laborious, time consuming, and determining the genotype of a CPV-2 variant often requires two or more reaction tubes. The present study developed a probe-based fluorescence melting curve analysis (FMCA) for genotyping six different CPV-2 variants (original CPV-2, CPV-2a, CPV-2b, CPV-2c, and vaccine strains of CPVpf and CPVint) in a single reaction tube using only two TaqMan probes. One of the TaqMan probes (FAM labeled) was designed to perfectly match with the target sequence of CPV-2a, this probe allows a 1-bp mismatched hybridization with the CPV-2b VP2 gene region (A4062G), and a 2-bp mismatched hybridization for CPV-2c (A4062G and T4064A); Another TaqMan probe (HEX labeled) was produced to perfectly match with the target sequence of original CPV-2, this probe enables 1-bp mismatched hybridization with the other CPV-2 variants (A3045T). Using the two TaqMan probes, all six CPV-2 variants were readily distinguished by their respective melting temperature values in a single reaction tube. The detection limits of this assay were 1–10 copies per reaction for six CPV-2 construction plasmids and no cross reactions were observed with several other common canine viruses. In this assay, co-infected samples were also directly identified via probe-based FMCA without using a mixing control; only a pure control is required. The clinical evaluation of this assay was demonstrated by analyzing 83 clinical fecal samples, among which 41 (49.39%), 8 (9.63%), and 14 (16.87%) samples were found to be positive for CPV-2a, CPV-2b, and CPV-2c, respectively. The concordance rate between probe-based FMCA and Sanger sequencing was 100%. Thus, the duplex FMCA is effective, rapid, simple, high-throughput, and straightforward for genotyping CPV-2 variants, and is useful to effectively diagnose and monitor CPV-2 epidemiology.

Genetic Characterization of Canine Parvovirus Type 2 Detected in Wild Raccoon Dogs ( Nyctereutes procyonoides) in the Republic of Korea.

Canine parvovirus (CPV) was detected in three of 136 samples from dead raccoon dogs ( Nyctereutes procyonoides) in the Republic of Korea (South Korea) during 2016-17. By sequence and phylogenetic analysis of the complete VP2 gene, the strain belonged to CPV-2 and would be distinct from the previous reported CPV-2a and CPV-2b strains from Korean domestic dogs ( Canis lupus familiaris). The results indicated that the CPV strains from raccoon dogs and domestic dogs might be not circulated between wild and domestic carnivores in Korea.

Molecular typing of a novel canine parvovirus type 2a mutant circulating in Italy

Canine parvovirus (CPV) is the etiological agent of a severe viral disease of dogs. After its emergence in late 1970s, the CPV original type (CPV-2) was rapidly and totally replaced by three antigenic variants named CPV-2a, CPV-2b and CPV-2c. CPV has an evolutionary rate nearest to those of RNA viruses, with consequences on disease diagnosis and epidemiology. This paper reports the molecular characterization of eight CPV-2a strains collected from dogs in Italy in 2016–2017. Genetic analysis was conducted on a CPV genomic region encompassing both open reading frames (ORFs) encoding for nonstructural (NS1-NS2) and structural proteins (VP1-VP2). Sequence analysis indicates new and unreported sequence changes, mainly affecting the VP2 gene, which included the mutation Tyr324Leu. This study represents the first evidence of a new CPV-2a mutant (VP2 324Leu) and illustrates the importance of a continuous molecular survey in order to obtain more information on effective spread of new CPV mutants.

Investigation on Canine parvovirus circulation in dogs from Sicily (Italy) by biomolecular assay

Abstract Canine parvovirus type-2 (CPV-2) is a DNA virus that causes a very common worldwide diffused infectious disease in dogs. Since its appearance, the CPV-2 evolved generating novel genetic and antigenic variants (CPV-2a/2b/2c) which are distributed throughout the world. In the present study, the frequency of CPV-2 in a canine population in Sicily (Italy) was investigated, using a polymerase chain reaction (PCR) for a fragment of the VP2 gene. Out of a total of 673 samples from 370 dogs, submitted to the laboratory from July 2009 to August 2015, 265 samples (39.38%) were positive and these were further analyzed by restriction fragment length polymorphism (RFLP) and DNA sequence analysis. A high prevalence of the CPV-2c variant (79.56% of CPV-2 positive dogs) was observed in this survey, underlining that this type of CPV field isolate is prevalent circulating in Sicily. Sequence and phylogenetic analysis showed a close relationship with CPV-2a and CPV-2c strains from Europe and non-European countries. Considering that CPV-2c is reaching a worldwide distribution and that this variant is also affecting vaccinated dogs, efforts should be made towards the development of new CPV vaccines.

Full-length genomic and molecular characterization of Canine parvovirus in dogs from North of Brazil.

With the objective of characterizing Canine parvovirus (CPV) from some suspected fecal samples of dogs collected from the Veterinarian Hospital in Belém city, five positive samples were found by PCR assay and an update molecular characterization was provided of the CPV-2 circulation in Belém. Through sequencing of the complete DNA sequences (NS1, NS2, VP1, and VP2 genes), the CPV-2 strain was identified as CPV-2b (Asn426Asp) circulating in Belém. The CPV-2b strain with a different change at the position Tyr324Leu was detected in all samples assessed and thus reported for the first time for the scientific community. Phylogenetic analysis indicated that Belém CPV-2b and CPV-2a strains would be related to a cluster with samples after the 1990s, suggesting that CPV-2b in Belém originated from CPV-2a circulating in Brazil after the 1990s. Potential recombination events were analyzed using RDP4 and SplitsTree4; therefore, results suggest that CPV-2 sequences here described were not potentially recombination events. Continuous monitoring and molecular characterization of CPV-2 samples are needed not only to identify possible genetic and antigenic changes that may interfere with the effectiveness of vaccines but also to bring a better understanding of the mechanisms that drive the evolution of CPV-2 in Brazil.

Genetic characterization of canine parvovirus type 2 subtypes in Maputo, Mozambique.

Canine parvovirus type 2 (CPV-2) comprises three antigenic subtypes (2a, 2b and 2c) that have been reported in many countries. These subtypes cause serious disease in dogs with characteristic gastroenteritis signs. Little information has been documented in Africa about the genetic characterization of CPV-2. The aim of this study was to detect and to characterize the CPV-2 subtypes circulating in dogs admitted to Veterinary Clinics from two cities of Mozambique, Maputo and Matola, in 2010. A total of 40 field fecal samples were collected and tested for CPV-2 by polymerase chain reaction assay. The partial length VP2 gene of the positive samples were sequenced and genetically analyzed. Twenty-six (65%) fecal samples were positive for CPV-2. The restriction fragment length polymorphism analysis was also performed from positive samples and did not reveal the presence of CPV-2c subtype. The results of the sequencing revealed the presence of CPV-2a (n=9) and CPV-2b (n=17). No CPV-2 and CPV-2c were detected. Sequence analysis comparison showed nucleotide identities of 99.6-100% among our CPV-2 isolates. Amino acid analysis showed predicted amino acid changes. Phylogenetically, all of the CPV-2a strains isolated formed a cluster together with South African and Nigerian isolates. Most of Mozambican CPV-2b isolates also tended to cluster together with South African isolates; however, four were more closely related to French strain and one isolates to the American strain. The present study was the first to characterize the CPV-2 circulating in the Mozambican dog population.

Molecular characterisation of parvoviruses from domestic cats reveals emergence of newer variants in India.

Objectives The present study was undertaken to characterise the viral polypeptide 2 (VP2) gene of parvovirus from domestic cats in India. Methods The faecal samples from diarrhoeic/healthy domestic cats were collected from different geographical regions of India for screening by PCR assay followed by sequence analysis of the VP2 gene. Results Canine parvovirus (CPV)/feline panleukopenia virus (FPV) infections were found in 12 (11.3%) of 106 faecal samples tested. Two new CPV-2a (297Ala and Asn426) and three FPV strains were identified by VP2 gene analysis. Several unique and existing amino acid mutations were found, suggesting continuous evolution and emergence of newer variants. The phylogenetic analysis of the CPV sequences revealed that the two new CPV-2a strains from Mumbai (MC8) and Puducherry (P15) were clustered together in a single clade but had evolved independently and were ancestrally related to Chinese CPV-2a isolates. The FPV sequences (T-C-6 and T-C-1) from Thrissur, Kerala, formed a different clade (FPV clade) and were closely related to each other and had an ancestral relationship with an FPV isolate from the USA. Another FPV isolate from Goa (GC1) was positioned in the same clade but had evolved independently. Conclusions and relevance Detection of CPV in both diarrhoeic/healthy cats and the occurrence of FPV infection in a vaccinated cat provide new insights into parvovirus infections in cats in India.

Full-length genomic characterization and molecular evolution of canine parvovirus in China.

Canine parvovirus type 2 (CPV-2) can cause acute haemorrhagic enteritis in dogs and myocarditis in puppies. This disease has become one of the most serious infectious diseases of dogs. During 2014 in China, there were many cases of acute infectious diarrhoea in dogs. Some faecal samples were negative for the CPV-2 antigen based on a colloidal gold test strip but were positive based on PCR, and a viral strain was isolated from one such sample. The cytopathic effect on susceptible cells and the results of the immunoperoxidase monolayer assay, PCR, and sequencing indicated that the pathogen was CPV-2. The strain was named CPV-NY-14, and the full-length genome was sequenced and analysed. A maximum likelihood tree was constructed using the full-length genome and all available CPV-2 genomes. New strains have replaced the original strain in Taiwan and Italy, although the CPV-2a strain is still predominant there. However, CPV-2a still causes many cases of acute infectious diarrhoea in dogs in China.