Canine Parvovirus Type Research Articles

Molecular Detection of Viral and Bacterial Pathogens in Red Foxes (Vulpes vulpes) from Italy.

Animals, including wildlife, are part of One-Health concept since many infectious diseases can affect both humans and animals. In this study, 126 red foxes (Vulpes vulpes) from Northern Italy in 2022-2023 were tested by molecular assays for Protoparvovirus carnivoran 1 (PPVC-1), Canine adenovirus type 1 and 2 (CAdV-1 and CAdV-2), Circovirus canine (CanineCV), Canine distemper virus (CDV), and Leptospira spp. A total of 39 of 126 (30.9%) red foxes were infected with at least one pathogen and five of these were coinfected: 20/126 (15.9%) red foxes tested positive for PPVC-1, 3/126 (2.4%) for CAdV, 20/126 (15.9%) for CanineCV, and 2/126 (1.6%) for Leptospira spp. DNA. No foxes tested positive for CDV RNA. The pathogens identified were genetically analysed. New findings were reported such as a fox with multiple feline panleukopenia virus (FPV) and canine parvovirus type 2b (CPV-2b) infection associated with quasispecies dynamics, typical genetic characteristics of the identified CanineCV, and the first detection in red foxes of Leptospira ST198 related to L. interrogans serogroup Australis. Further studies are necessary to investigate the transmission between domestic animals and wildlife and to understand the role of red foxes in the maintenance of these pathogens not only in the wild but also in urban and peri-urban environments.

First identification of canine parvovirus -2a/2b variant in unvaccinated domestic dogs with gastrointestinal signs in Türkiye.

Canine parvovirus type 2 (CPV-2) is the most common enteric virus that infects canids. CPV is the causative agent of a contagious disease defined mostly by clinical gastrointestinal signs in dogs. During the late 1970s, CPV-2 emerged as a new virus capable of infecting domestic dogs and growing across the world. The VP2 gene stands out as a key determinant in the pathogenicity, antigenicity, and host interactions of CPV-2. The molecular characterization of the VP2 gene is crucial for understanding CPV evolution and epidemiology. Genes encoding the VP2 protein were sequenced and compared to reference strains worldwide. The maximum likelihood method was used to build a phylogenetic tree using CPV VP2 gene nucleotide sequences. Our phylogenetic analysis of the VP2 gene revealed that five strains were very similar and clustered together, and three strains were in the 2b clade, whereas the other two were in the 2a/2b clade. This paper reports the molecular characterization of two novel CPV-2a/2b subtypes in dogs with gastrointestinal symptoms. Genetic analysis was conducted on a CPV genomic region encompassing one of the open reading frames (ORFs) encoding the structural protein VP2. Sequence analysis indicates new and unreported sequence changes, mainly affecting the VP2 gene, which includes the mutations Ser297Ala and Leu87Met. This study represents the first evidence of a new CPV-2a/2b subtype in Türkiye. Due to VP2's crucial role in encoding the capsid protein of CPV-2 and its significant involvement in the host-virus interaction, it is critical to closely monitor its evolutionary changes and be cautious while searching for novel or pre-existing subtypes. This study highlights the significance of continuous molecular research for acquiring more insights on the circulation of novel CPV mutants.

Viral pathogens in domestic cats in southern Italy: A retrospective analysis in Sicily, 2020–2022

A retrospective study was carried out on selected feline viral pathogens detected in domestic cat in Sicily, southern Italy. Samples from 64 cats, collected from 2020 to 2022, were analysed for the presence of feline panleukopenia virus, canine parvovirus type 2 (CPV-2), feline coronavirus (FCoV), feline calicivirus (FCV), feline herpesvirus type 1, norovirus (NoV), and rotavirus (RoV). Single (45 %) or mixed (38 %) viral infections were detected. FPV, related with other Italian FPV strains, remains the main viral cause of infection (66 %). CPV-2c Asian lineage strains (3 %) were detected for the first time in domestic cats in Europe. FCoV (29.6 %), either enteric or systemic, and systemic FCV (18.7 %) infections were detected in positive cats. Less commonly reported viruses (GIV.2/GVI.2 NoVs, RoV), potentially related to the animal/human interface, were detected at lower rates as well (5 %). The present epidemiological data suggest the need to improve disease prevention, immunization, and biosecurity strategies.

OVERVIEW OF THE CANINE PARVOVIRUS TYPE 2: ETIOLOGY, PATHOGENESIS, DIAGNOSIS, TREATMENT, AND PREVENTION

Parvo in dogs is caused by canine parvovirus type 2 (CPV-2), a highly contagious viral disease in dogs that often causes high mortality and is mainly transmitted through the gastrointestinal tract. Nowadays, the variants identified include CPV-2a, CPV-2b, and CPV-2c. This pathogen has appeared in most countries around the world, including Vietnam. Its host range varies with each type of variant, and its relative frequency and genetic variability also vary between countries. The research provides an overview of key information regarding themolecular structure, origin, evolution, and pathophysiology of CPV-2, drawing on 30 published study results from various countries worldwide. This paper helps to identify risk factors that contribute to CPV-2 disease. In addition, the research also synthesizes research on advanced diagnosis, treatment, and CPV-2 vaccine development, serving as a basis for veterinarians, veterinary students, and breeders to control and limit the impact of upper enteritis caused by CPV-2.

Detection of canine parvovirus type 2c (CPV-2c) in Palestine.

The objective of the present study was to report, for the first time, the presence of canine parvovirus type 2c (CPV-2c) in domesticated dogs with acute gastroenteritis and to characterize the antigenic variants circulating in Palestine. A veterinary clinical-based epidemiological study was carried out between December 2022 and April 2023. Fifty fecal samples were collected from dogs with gastroenteritis and screened for CPV-2 infection by polymerase chain reaction. The distribution of positive cases according to various epidemiological factors was studied. Partial sequencing of the viral protein 2 (VP2) gene was performed for the analysis of CPV-2 variants. Most of the investigated samples (60%; n = 50) during the study period were found positive for CPV-2 infection. There was no difference in the distribution of positive cases of CPV-2 infection based on age group, gender, location, and vaccination status. The analysis of nucleotide and amino acid sequences from amplified products, as well as phylogenetic analysis, revealed the presence of CPV-2c clustered with Asian CPV-2c variants. In summary, this study represents the initial genetic analysis of CPV-2 present in Palestinian dogs with gastroenteritis and provides evidence that confirms the existence of the CPV-2c variants. To determine the prevailing CPV-2 variant associated with the infection, it is crucial to conduct further sequence analysis using large populations of both domestic and wild canines.

Molecular characterization of full-length VP2 gene of canine parvovirus type 2 strains circulating in Egypt 2019–2021

Canine parvovirus type 2 (CPV-2) is a major cause of fatal gastroenteritis and myocarditis in puppies of domestic and wild carnivores. CPV-2 has accumulated changes over time lead to the emergence of three antigenic variants CPV-2a, CPV-2b, and CPV-2c. VP2 is the major capsid protein that determines virus antigenicity, and host range. Although the three CPV-2 variants were previously identified in Egypt, most reports covered a restricted geographic region and/or time period, and only analyzed partial fragments of VP2 gene. Therefore, this study was designed to test 100 rectal swabs collected from 7 Egyptian governorates between 2019 and 2021 for CPV-2 using PCR. A total of 65 positive samples were identified, mostly in pure dog breeds of young age. The three variants co-circulated in 2019, while CPV-2b was not detected in 2020 and 2021. The frequency of CPV-2b and CPV-2c was higher in 2019 and 2021, respectively. Analysis of CPV-2 full-length VP2 gene sequence from 19/65 positive samples has identified four common amino acid substitutions F267Y, S297A, A300G, Y324I, which are characteristic for the new CPV-2 variants currently circulating worldwide. Unique substitutions including A5G, G36R, V38E, Q370R, and G392V were recognized in certain samples, and appears to have distinct effect on receptor binding, nuclear translocation, and inter-species transmission. Phylogenetic analysis showed separation of CPV-2 strains into two clades. All strains of this study were classified in clade I with Asian strains. In conclusion, this study provides updated comprehensive molecular analysis of CPV-2 variants in Egypt.

Investigation of canine parvovirus occurrence in cats with clinical signs of feline panleukopenia in Slovakia - pilot study.

Feline panleukopenia is a contagious viral disease caused by the feline panleukopenia virus (FPV). A closely related pathogen is canine parvovirus (CPV), and amino acid substitutions in this virus allow it to acquire a feline host range. In feline hosts, the disease induced by CPV manifests with similar symptoms to those caused by FPV or milder ones, leading to its underdiagnosis. The aim of this study was to determine the presence of CPV type 2 (CPV-2) in cats with clinical symptoms of panleukopenia and to assess the use of commercial CPV antigen tests for the clinical diagnosis of FPV. Samples from 59 cats from central Slovakia were included in the study. Rectal swabs were collected and clinically tested for parvovirus infection using a commercial antigen test. Antigen-positive samples were confirmed by PCR targeting the viral VP2 gene. The sequences of the PCR products were established with the Sanger method. Of 59 samples, 23 were revealed to be positive for parvovirus infection by both antigen and PCR test (38.9%). Analysis with the National Center for Biotechnology Information BLASTn application showed 99.78-100% pairwise identity with FPV. The mortality rate of parvovirus-infected cats included in this study was 8.69% (2/23). Although feline disease with CPV-2 was not confirmed, the CPV antigen test was able to detect FPV infection.

Global distribution, cross-species transmission, and receptor binding of canine parvovirus-2: Risks and implications for humans

For canine parvovirus -2 (CPV-2), a zoonotic virus capable of cross-species transmission in animals, the amino acid changes of capsid protein VP2 are key factors when binding to other species' transferrin receptors (TfR). CPV-2 variants can spread from felines and canines, for example, to Carnivora, Artiodactyla, and Pholidota species, and CPV-2c variants are essential to spread from Carnivora to Artiodactyla and Pholidota species in particular. In our study, a CPV-2a variant maintained a relatively stable trend, and the proportion of CPV-2c gradually rose from 1980 to 2021. The VP2 amino acid sequence analysis showed that five amino acid mutations at 426E/D, 305H/D, and 297S may be necessary for the virus to bind to different host receptors. Meanwhile, receptor-binding loop regions and amino acid sites 87 L, 93 N, 232I, and 305Y were associated with CPV-2 cross-species transmission. The homology of TfRs in different hosts infected with CPV-2 ranged from 77.2 % to 99.0 %, and from pig to feline, canine, and humans was 80.7 %, 80.4 %, and 77.2 %, respectively. The amino acid residues of TfRs involved in the viral binding in those hosts are highly conserved, which suggests that CPV-2 may be capable of pig-to-human transmission. Our analysis of the origin, evolutionary trend, cross-species transmission dynamics, and genetic characteristics of CPV-2 when binding to host receptors provides a theoretical basis for further research on CPV-2's mechanism of cross-species transmission and for establishing an early warning and monitoring mechanism for the possible threat of CPV-2 to animal–human public security.

Genome Sequences of Canine Parvovirus Type 2c Prevalent in Western Mexico.

Canine parvovirus type 2 (CPV-2) is one of the main etiologies of viral gastroenteritis in dogs across the globe. This disease is mainly characterized by the presence of diarrhea, abdominal pain, vomiting, anorexia, and dehydration. This virus is responsible for high mortality and morbidity rates in unvaccinated dogs and those younger than three months. The monitoring of viral variants in our region has demonstrated that in the last seven years, variant CPV-2c has been circulating exclusively, which is unusual if we consider that in the rest of the world, at least two variants co-circulate among dog populations. To the best of our knowledge, no studies in Mexico have reported genomic sequences of CPV-2,which are relevant for population comparisons at the genetic level. Therefore, the present study aimed to sequence genomes associated with CPV-2c. To meet this objective, rectal swab samples were collected from dogs with suspected CPV-2 infection. Five positive cases diagnosed by lateral flow testing and polymerase chain reactionwere selected for viral genome sequencing. Comparative analyses illustrated that the obtained genome sequences were > 99% homologous to those reported for CPV-2 in the GenBank. On the other hand, 52 nucleotide mutations were identified in thevp1/vp2gene, out of which three impacted amino acid transition (T226S, F267Y, and A440T). Phylogenetic analysis of thevp1/vp2gene demonstrated that the five sequences clustered in a clade called "III", pertaining to sequences from USA and Uruguay. To our knowledge, this was the first report of genomic sequences associated with CPV-2 in Mexico, which is of great relevance for the epidemiological-molecular understanding and evolution of the virus.

Development of a time-resolved fluorescence immunoassay kit for detecting canine coronavirus and parvovirus through double labeling

ObjectiveCanine enteric coronavirus (CCV) and canine parvovirus type 2 (CPV-2) are the main pathogens responsible for acute gastroenteritis in dogs, and both single and mixed infections are common. This study aimed to establish a double-labeling time-resolved fluorescence immunoassay (TRFIA) to test and distinguish CCV and CPV-2 diseases.MethodsA sandwich double-labeling TRFIA method was established and optimized using europium(III) (Eu3+)/samarium(III) (Sm3+) chelates. CCV/CPV-2 antigens were first captured by the immobilized antibodies. Then, combined with Eu3+/Sm3+-labeled paired antibodies, the Eu3+/Sm3+ fluorescence values were detected after dissociation to calculate the CCV/CPV-2 ratios. The performance, clinical performance and methodology used for laboratory (sensitivity, specificity, accuracy and stability) testing were evaluated.ResultsA double-label TRFIA for CCV and CPV-2 detection was optimized and established. The sensitivity of this TRFIA kit was 0.51 ng/mL for CCV and 0.80 ng/mL for CPV-2, with high specificity for CCV and CPV-2. All the accuracy data were less than 10%, and the recovery ranged from 101.21 to 110.28%. The kits can be temporarily stored for 20 days at 4 °C and can be stored for 12 months at temperatures less than − 20 °C. Based on a methodology comparison of 137 clinically suspected patients, there was no statistically significant difference between the TRFIA kit and the PCR method. Additionally, for CCV detection, the clinical sensitivity was 95.74%, and the clinical specificity was 93.33%. For CPV-2 detection, the clinical sensitivity was 92.86%, and the clinical specificity was 96.97%.ConclusionIn this study, a double-label TRFIA kit was prepared for CCV and CPV-2 detection with high laboratory sensitivity, specificity, accuracy, stability, clinical sensitivity and specificity. This kit provides a new option for screening/distinguishing between CCV and CPV-2 and may help improve strategies to prevent and control animal infectious diseases in the future.

Canine parvovirus type 2 (CPV-2) serological and molecular patterns in dogs with viral gastroenteritis from southern Brazil.

Canine Parvovirus type 2 (CPV-2) is a highly contagious virus that can cause severe systemic disease with gastroenteric symptoms in dogs, particularly in young puppies. Originating from the feline parvovirus in the late 1970s, it swiftly propagated globally, instigating a pandemic in dogs. Despite vaccination advancements, CPV-2 remains a substantial challenge for veterinary professionals and pet owners. This study aimed to contribute knowledge about the current situation of CPV-2 among dogs in southern Brazil. In this study, the sera of 125 dogs (mostly with gastroenteritis symptoms) were screened for antibodies against CPV-2 and their faeces for the virus itself. The results showed that 40% (50/125) of dogs were infected with CPV-2. Most animals (65.5%) had previously been exposed to CPV-2 (with serotitres equal or above 1:40), and only 37.6% had protective antibody titres equal or above 1:80. The findings have also demonstrated that vaccination against CPV-2 significantly reduced the risk of infection, with positive cases decreasing from 56.9% (unvaccinated) to 2.0% (fully vaccinated). Furthermore, the prevalence of CPV-2 decreased as dogs aged, with younger dogs and those with an incomplete or non-existent vaccination history at the highest risk of infection. In conclusion, this study provides valuable insight into the prevalence and risk factors associated with CPV-2 infection in dogs in southern Brazil, thereby providing valuable knowledge for the improvement of veterinary care and pet health.

Antiviral alternatives against important members of the subfamily Parvovirinae: a review.

Parvoviruses are responsible for multiple diseases, and there is a critical need for effective antiviral therapies. Specific antiviral treatments for parvovirus infections are currently lacking, and the available options are mostly supportive and symptomatic. In recent years, significant research efforts have been directed toward understanding the molecular mechanisms of parvovirus replication and identifying potential targets for antiviral interventions. This review highlights the structure, pathogenesis, and treatment options for major viruses of the subfamily Parvovirinae, such as parvovirus B19 (B19V), canine parvovirus type 2 (CPV-2), and porcine parvovirus (PPV) and also describes different approaches in the development of antiviral alternatives against parvovirus, including drug repurposing, serendipity, and computational tools (molecular docking and artificial intelligence) in drug discovery. These advances greatly increase the likelihood of discoveries that will lead to potent antiviral strategies against different parvovirus infections.

Molecular epidemiology of canine parvovirus type 2 in Sicily, southern Italy: A geographical island, an epidemiological continuum

Since it emerged as a major dog pathogen, canine parvovirus type 2 (CPV-2) has featured a remarkable genetic and phenotypic heterogeneity, whose biological, epidemiological, and clinical impact is still debated. The continuous monitoring of this pathogen is thus of pivotal importance. In the present study, the molecular epidemiology of CPV-2 in Sicily, southern Italy, has been updated by analysing 215 nearly complete sequences of the capsid protein VP2, obtained from rectal swabs/faeces or tissue samples collected between 2019 and 2022 from 346 dogs with suspected infectious gastrointestinal disease. The presence of the original CPV-2 type (4%) and CPV-2a (9%), CPV-2b (18%), or CPV-2c (69%) variants was documented. Over the years, we observed a decrease in the frequency of CPV-2a/-2b and a rapid increase of CPV-2c frequency, with a progressive replacement of the European lineage of CPV-2c by the Asian lineage.The observed scenario, besides confirming epidemiological relevance of CPV-2, highlights the occurrence of antigenic variant shifts over time, with a trend toward the replacement of CPV-2a, CPV-2b, and the European lineage of CPV-2c by the emerging Asian CPV-2c lineage. The comparison with other Italian and international sequences suggests the occurrence of viral exchange with other Italian regions and different countries, although the directionality of such viral flows could not be often established with confidence. In several instances, potential CPV-2 introductions led to epidemiological dead ends. However, major, long-lasting clades were also identified, supporting successful infection establishment, local spreading, and evolution. These results, besides demonstrating the need for implementing more effective control measures to prevent viral introductions and minimize circulation, stress the relevance of routine monitoring activities as the only tool to effectively understand CPV-2 epidemiology and evolution, and develop adequate countermeasures.

CANINE PARVOVIRUS INFECTION IN DOGS: PATHOGENESIS, CLINICAL SYMPTOMS, DIAGNOSIS, PREVALENCE, THERAPY, AND VACCINATION

Canine parvovirus is a highly contagious disease in dogs that attacks the digestive tract. This disease is caused by canine parvovirus type 2 (CPV-2), belonging to the Parvoviridae family. This virus causes the highest mortality in dogs under six months of age. This virus has a very high mortality and morbidity rate, especially in dogs that have not been vaccinated. This literature review discusses the pathogenesis of CPV, the clinical symptoms that can be caused, how to diagnose, the prevalence of CPV cases, therapy, and vaccination to prevent transmission of canine parvovirus. The method used is literature study by collecting and summarizing references relevant to the topic. Based on this literature review, the results obtained include that canine parvovirus infection can occur directly or indirectly and CPV will show various clinical symptoms, namely the type of myocarditis and the type of enteritis. Common methods used to help confirm a diagnosis include anatomical pathology examinations and laboratory diagnostic examinations in the form of blood hematology examinations, test kits, Polymerase Chain Reaction (PCR), Fluorescent Antibody Technique (FAT), Enzyme Linked Immunosorbent Assay (ELISA). Based on the prevalence of CPV cases, it is known that CPV mostly attacks dogs aged 0-6 months. Therapy for CPV sufferers is only symptomatic and supportive. Vaccination is the main way to avoid CPV disease. It is hoped that more research on CPV will be carried out so that insight into CPV is easier to obtain and can become a lesson for the future.

Prevalence and Molecular Evolution of Parvovirus in Cats in Eastern Shandong, China, between 2021 and 2022

Feline panleukopenia (FPL) is a highly contagious infectious disease caused by infection with feline parvovirus (FPV) and canine parvovirus type 2 (CPV-2). In recent years, the number of cats with FPL has increased with the expansion of pet cat population in China. The feces of 51 cats with diarrhea symptoms collected from 2021 to 2022 in Eastern Shandong, China, were detected by polymerase chain reaction for parvovirus and other viruses related to feline diarrhea to investigate the prevalence and gene variation of parvovirus in cats. In all the 51 samples, 45.1% (23/51) were positive for at least one viral pathogen, and the positivity of parvovirus was 41.2% (21/51), showing a high prevalence. Multiple-pathogen testing indicated high-coinfection rates of 42.9% (9/21) with other common viruses in parvovirus-positive cats. Most of the coinfections are feline coronavirus (FCoV), followed by feline astrovirus (FAstV) and feline bocavirus (FBoV). The complete VP2 sequences of 21 parvoviruses were obtained. Among them, 20 sequences were identified as FPV, and only one was CPV-2c of Asian origin, which was first detected from cats in Eastern Shandong, China. A phylogenetic tree of the 20 FPVs was constructed together with 698 FPVs (cat/dog host) worldwide on the basis of complete VP2. The 18 FPVs displayed high-sequence identity to one another (99.8%–100%), and they were clustered into FPV-G1 group, whereas the other two were clustered into FPV-G3 group. The FPV-G1 group increased dramatically to become predominant after 2019 in China, contributing to the prevalence of A91S mutation due to 96.07% FPV-G1 with A91S mutation as well as 100% of FPV-G2 and 99.12% of FPV-G3 with 91A in the statistical analysis. This study enriched the understanding of the prevalence, molecular evolution, and cross-species transmission of parvovirus in cats and provided a basis for responding to challenges in the diagnosis and treatment of FPL.

Molecular characterization of canine parvovirus type 2 (CPV2) reveals a high prevalence of the CPV2c genotype among dogs suffering from diarrhea

AbstractCanine parvovirus 2 (CPV-2) is a highly contagious virus in dogs that typically causes hemorrhagic enteritis and a high mortality rate in unvaccinated puppies. The genetic variability and antigenic diversity of CPV-2 hinder its effective prevention of infection by vaccination. To investigate the epidemiology and genetic characteristics of CPV-2 in China, rectal swabs from affected dogs were collected from different animal clinics in Kunshan from 2022 to 2023. Preliminary detection and capsid gene sequencing of CPV-2 were performed using previously described primers and protocols. The overall detection rate for CPV-2 was 16.5% (33/200). A significant association was found between the CPV-2-positivity and clinical signs, age, breed and vaccination status. Sequence analysis revealed the presence of CPV-2c genotypes in all positive samples, which were genetically similar to other Asian CPV-2c strains. Notably, four key mutations (A5G, F267Y, Y324I and Q370R) were detected in all isolates, and one novel mutation (I447M) was detected in three CPV-2 isolates. These mutations in the CPV-2 strains could impact vaccine efficacy and the effectiveness of the virus immune evasion. Surprisingly, no recombination events were observed between the identified CPV-2c strains and reference strains from China. Our data revealed that amino acid residues 324, 426 and 440 of VP2 may under strong selection pressure. This pattern of genetic variation in the CPV-2 lineage warrants continuous laboratory-based surveillance programs in other parts of China to better understand the pattern of seasonal distribution and association between emerging genotypes and the intensity of disease severity.

Haematological changes in canine parvovirus infection

Canine parvovirus (CPV) infection is a highly contagious disease caused by canine parvovirus type 2 and commonly produce acute gastrointestinal illness. All dogs are susceptible to CPV, although some dogs are at greater risk than others, as puppies (between 6 weeks of age and 6 months), unvaccinated or incompletely vaccinated dogs. Due to the virus multiplication and effect on the bone marrow, severe haematological changes are reported, resulted also from the combination of severe inflammation, gastrointestinal bleeding and depletion of hematopoietic cell lines. In the present study, haematological changes in dogs diagnosed with CPV infection were analysed.

Molecular typing of canine parvovirus type 2 by VP2 gene sequencing and restriction fragment length polymorphism in affected dogs from Egypt.

Canine parvovirus-2 (CPV-2) is one of the most common infectious diseases in dogs characterized by severe gastroenteritis, vomiting, and bloody diarrhea. Little information is available about this topic in Egypt, particularly in the Delta region. This study reports the prevalence and molecular analysis of CPV-2 variants collected from El-Gharbia and Kafrelsheikh governorates in the Delta of Egypt. In this study, 320 rectal swabs were collected from infected domestic dogs from two districts in delta Egypt. The samples were investigated by rapid immunochromatographic test and polymerase chain reaction for detection the prevalence of CPV-2 variants. The genetic characterization was performed using restriction fragment length polymorphism (RFLP) analysis and partial VP2 gene sequence. The viral antigen was detected in (264/320, 82.5%) of samples by IC test, while PCR was found more sensitive by detecting (272/320, 85%) positive samples. The RFLP technique using MboII restriction enzyme was successfully used for the differentiation of CPV-2c antigenic variants from CPV-2a/2b strains. Interestingly, the molecular and phylogenetic analysis revealed that both CPV-2a and CPV-2c are circulating in the study area. Deduced amino acid sequence analysis showed changes at residue (N426E) and residue (T440A).: Our results indicated that CPV-2 is prevalent among dogs in Egypt, and therefore further molecular and epidemiological studies of CPV-2 are warranted.

Noticeable immune dysregulation-and-suppression in parvovirus affected dogs

Canine parvovirus type 2 (CPV-2) is one of the most common causes of infectious diarrhea in small animals, with high mortality and morbidity. Information on the specific treatment option(s) for CPV diseases (CPVD) is unachievably little. So, the treatment is mainly supportive one. Disruption of dog's innate immune system in viral diseases simply occurs; presumably, the CPV-2 may change the level of some TLRs, interleukins, CD4 and CD8 in the leukocytes of CPVD dogs, and disruptive activities of these immune molecules might be attributable to severe CPVD in dogs. Study on the role of the key immune molecules in CPVD is rare. Herein, by conducting and relating the clinical, para-clinical, immunological and molecular diagnostic tests, we tried to establish how some key immune molecules behave in blood of parvovirus affected dogs. As such, in the 1st study, the mRNA levels of TLR2, TLR4, TLR9, IL-1β, IL-6, CD4 and CD8 genes in the leukocytes of CPVD were assessed with quantitative (q)RT-PCR along with CPV-2 detection by rapid immunochromatography and PCR tests. In a 2nd study, the same measurements as in the 1st study were evaluated in two groups of mild versus severe clinical signs of CPVD. Both in the 1st and the 2nd studies leukopenia, much more pronounced in the severe CPVD, and immune dysregulation were observed. In the 1st study, a noticeable increase in the mRNA levels of TLR2 and TLR4 was detected with a slight decrease in TLR9 and a significant decrease in the expression of IL-1β, IL-6, CD4 and CD8 in leukocytes of CPV-infected dogs. Compared to the mild CPVD, the intense of downregulating effects on those immune molecules in the 2nd study was remarkably much more pronounced in the severe CPVD. Overall, it proves strong immune dysregulation and suppression/incompetence and potential T-cells exhaustion in severely CPV-2-affected dogs. Technically and clinically, this would be substantially applicable in canine medicine. By targeting those key immune molecules and their signaling pathways, new clinicodiagnostic approaches for CPVD can be evolved, and biotechnicoclinically this would be substantially applicable in all physiopathological conditions of dogs.

First report of detection of Canine Parvovirus type 2 in naturally infected domestic cats in Egypt by duplex PCR for simultaneous detection of Canine Parvovirus type 2 and Feline Panleukopenia virus

Feline Panleukopenia virus (FPLV) and Canine Parvovirus type 2 (CPV) cause fatal gastroenteritis in cats and dogs. In this study we developed a duplex polymerase chain reaction (dPCR) assay for the concurrent detection of FPLV and CPV-2 in a single PCR tube. Two primers were used based on nucleic acid conserved regions of the two viruses which specifically amplify 237 bp of the VP2 gene of FPLV and 583 bp of the VP2 gene of CPV 2.Sensitivity and Specificity of the dPCR were evaluated. A total of 30 rectal/fecal swabs were collected from domestic cats in Kafrelsheikh province, Egypt and were tested for FPLV and CPV-2 viruses using the dPCR assay. The results revealed that this dPCR assay was sensitive, as it could detect a minimum of 1 × 105 copies of genomic DNA of the two viruses. The dPCR assay was highly specific as there was no amplification of nucleic acid of other feline and canine pathogens. The positive ratio was 83.3% (25/30) for FPLV and 16.6% (5/30) for CPV respectively. Further analyses of CPV samples by Restriction Fragment Length Polymorphism (RFLP) revealed that they are classified as CPV 2a/2b variants. This study reports the first detection of CPV 2a/2b from symptomatic cats in Egypt using dPCR assay that can detect FPLV and CPV in a single tube reaction.