Caracterização Molecular do Parvovírus Canino tipo 2 e do vírus da Panleucopenia Felina em cães e gatos da Área Metropolitana de Lisboa
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2025
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O género Protoparvovirus engloba várias espécies, incluindo o parvovírus canino do tipo 2 (CPV-2) e o vírus da panleucopenia felina (FPV). Adicionalmente, o CPV-2 apresenta uma taxa de mutação nucleotídica frequente, levando a alterações aminoacídicas na proteína viral 2 (VP2), responsáveis pela transmissão, infeção e propriedades antigénicas deste vírus, traduzindo-se na existência de subtipos (CPV-2a, CPV-2b, CPV-2c). Neste contexto, pretendeu-se com o presente trabalho, fazer a caracterização molecular do CPV-2 e o FPV de cães e gatos infetados na área metropolitana de Lisboa e determinar a frequência de cada subtipo do CPV-2. Para tal, foram estudadas amostras fecais/sangue de 27 animais com sinais clínicos compatíveis com infeção causada por CPV-2 e FPV e com teste de antigénio (SNAP) positivo (Idexx Laboratories ®, Portugal). Procedeu-se à extração de DNA presente em amostras de fezes e sangue, posteriormente, procedeu-se à amplificação por PCR do gene vp2 e sequenciação dos produtos de amplificação. As sequências obtidas foram alinhadas com o programa MAFFT e a subtipificação viral com o programa Aliview. Posteriormente, realizou-se uma árvore filogenética e análise de haplótipos do CPV-2, do FPV e de comparação do vírus presente nas fezes e no sangue de seis amostras aleatórias. No total, o vírus mais prevalente na amostra foi FPV (48,0%). Nos gatos detetou-se FPV (93,0%) e CPV-2c (7,0%). Nos cães, o subtipo predominante foi o CPV-2c (77,0%), seguido do subtipo CPV-2b (23,0%). Foram identificadas algumas mutações nas amostras em estudo. No estudo da árvore filogenética, o CPV e o FPV demonstraram ser monofiléticos. Relativamente ao CPV-2 este demonstrou a presença de segregação de intragrupos, porém, nenhum destes refletiu monofilia para os subtipos. Estes resultados reforçam a questão da importância da identificação e monitorização dos subtipos do CPV-2 em circulação nas populações caninas e felinas para a compreensão da evolução do vírus, a sua epidemiologia e o seu impacto na saúde dos hospedeiros. Estes apoiam a necessidade de análises moleculares regulares e de uma revisão da classificação com base na sequência do genoma completo do subtipo viral.
The genus Protoparvovirus comprises several species, including canine parvovirus type 2 (CPV-2) and feline panleukopenia virus (FPV). Additionally, CPV-2 exhibits a high nucleotide mutation rate, leading to amino acid changes in the Viral Protein 2 (VP2), which are responsible for the virus's transmission, infection, and antigenic properties, resulting in the existence of subtypes (CPV-2a, CPV-2b, CPV-2c). In this context, the present study aimed to characterise CPV-2 and FPV from infected dogs and cats in the Lisbon Metropolitan Area (LMA), to determine the prevalence of CPV-2 subtype through nucleotide sequencing. A total of 27 animals presenting clinical signs compatible with CPV-2 and FPV infections were tested, all of which had a positive result in an antigen (SNAP) test (Idexx Laboratories®, Portugal). DNA was extracted from faecal and blood samples, followed by PCR amplification of the vp2 gene and sequencing of the amplification products. The obtained sequences were aligned using the MAFFT programme, and viral subtyping was performed using the Aliview programme. Subsequently, a phylogenetic tree and haplotype analysis were constructed to compare CPV-2 and FPV isolates as well as the virus present in faeces and blood from six samples. Overall, the most prevalent virus was FPV (48,0%). In cats, FPV was detected in 93,0% of cases, and CPV-2c in 7,0%. In dogs, the predominant subtype was CPV-2c (77,0%), followed by CPV-2b (23,0%). Several mutations were identified in the analysed samples. Phylogenetic tree analysis demonstrated that CPV and FPV were monophyletic. Regarding CPV-2, intragroup segregation was observed; however, none of the clusters exhibited monophyly for subtypes These findings emphasise the importance of identifying and monitoring circulating CPV-2 subtypes in canine and feline populations to better understand the virus's evolution, epidemiology, and its impact on host health. This study supports, the need for regular molecular analyses and a revision of the classification based on the full genome sequence of the viral subtype.
The genus Protoparvovirus comprises several species, including canine parvovirus type 2 (CPV-2) and feline panleukopenia virus (FPV). Additionally, CPV-2 exhibits a high nucleotide mutation rate, leading to amino acid changes in the Viral Protein 2 (VP2), which are responsible for the virus's transmission, infection, and antigenic properties, resulting in the existence of subtypes (CPV-2a, CPV-2b, CPV-2c). In this context, the present study aimed to characterise CPV-2 and FPV from infected dogs and cats in the Lisbon Metropolitan Area (LMA), to determine the prevalence of CPV-2 subtype through nucleotide sequencing. A total of 27 animals presenting clinical signs compatible with CPV-2 and FPV infections were tested, all of which had a positive result in an antigen (SNAP) test (Idexx Laboratories®, Portugal). DNA was extracted from faecal and blood samples, followed by PCR amplification of the vp2 gene and sequencing of the amplification products. The obtained sequences were aligned using the MAFFT programme, and viral subtyping was performed using the Aliview programme. Subsequently, a phylogenetic tree and haplotype analysis were constructed to compare CPV-2 and FPV isolates as well as the virus present in faeces and blood from six samples. Overall, the most prevalent virus was FPV (48,0%). In cats, FPV was detected in 93,0% of cases, and CPV-2c in 7,0%. In dogs, the predominant subtype was CPV-2c (77,0%), followed by CPV-2b (23,0%). Several mutations were identified in the analysed samples. Phylogenetic tree analysis demonstrated that CPV and FPV were monophyletic. Regarding CPV-2, intragroup segregation was observed; however, none of the clusters exhibited monophyly for subtypes These findings emphasise the importance of identifying and monitoring circulating CPV-2 subtypes in canine and feline populations to better understand the virus's evolution, epidemiology, and its impact on host health. This study supports, the need for regular molecular analyses and a revision of the classification based on the full genome sequence of the viral subtype.
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MESTRADO INTEGRADO EM MEDICINA VETERINÁRIA, VETERINÁRIA, MEDICINA VETERINÁRIA, GATOS, FELÍDEOS, CÃES, CANÍDEOS, FILOGENIA, VIROLOGIA ANIMAL, EPIDEMIOLOGIA VETERINÁRIA, ÁREA METROPOLITANA DE LISBOA, VETERINARY MEDICINE, CATS, FELIDS, DOGS, CANIDS, PHYLOGENY, VETERINARY VIROLOGY, ANIMAL EPIDEMIOLOGY, LISBON METROPOLITAN AREA