Spatiotemporal expression pattern of alpha, beta, and gamma genes during BoHV-5 infection / Expressão temporal de genes alfa, beta e gama durante ainfecção pelo BoHV-5

Bovine herpesvirus 5 is an alphaherpesvirus that causes nonsuppurative meningoencephalitis in cattle. This disease occurs naturally in either outbreaks or isolated cases, and exhibits low morbidity and high lethality. Although previous studies elucidated crucial aspects involved in the pathogenesis of the disease, there is a paucity of information regarding the molecular events contributing to infection and replication of BoHV-5. The objective of the present study was to determine the in vitro gene expression pattern of BoHV-5 (e.g., alpha, beta, and gamma genes) and host cells genes (GAPDH and 18S) over time utilizing different quantities of inoculated virus. Three BoHV-5 genes (bICP0, UL9, US4) and one structural bovine cell gene had their expression accessed by real-time PCR. While the expression of BoHV-5 genes increased during the course of infection, GAPDH gene expression decreased in the host cells, evidencing the effect of viral infection on the expression of bovine cell genes. The 18S ribosomal RNA (rRNA) gene was constitutively expressed throughout BoHV-5 infection. Our data clearly demonstrates that GAPDH gene should not be used as a reference gene in studies of BoHV-5 infection because it was influenced by viral infection. However, 18S rRNA was constitutively expressed and, therefore, is recommended for normalization of BoHV-5 infection studies in bovine cells. The expression of viral genes transcripts was not altered by increasing number of viral particles added to the culture. All viral genes included here demonstrated the same expression pattern over time and there was no difference in the expression of viral genes among the various time points. Our data show important differences comparing to classical studies regarding herpesvirus alpha, beta, and gamma genes expression. More research is necessary to improve our understanding about the BoHV-5 biology during infection. Studies employing next-generation sequencing (i.e., RNA-seq), using both in vitro and in vivo models, would be the next logical step to grasp the virus and host cell's transcriptome changes over the course of infection.(AU)

Herpesvirus bovino 5 é um alfaherpesvírus causador de meningoencefalite não supurativa em bovinos. Esta doença possui ocorrência natural em surtos ou casos isolados, associadas a baixa morbidade e alta letalidade. Embora estudos anteriores tenham elucidado aspectos relacionados a patogenia da doença, há uma lacuna de conhecimento relacionado aos eventos moleculares que contribuem para a infecção e replicação do BoHV-5. O objetivo do presente estudo foi determinar a expressão gênica in vitro de genes virais (i.e., alfa, beta e gama) e das células hospedeiras (GAPDH e 18S) durante a infecção considerando diferentes momentos de infecção e quantidade de vírus utilizado. Três genes do BoHV-5 (bICP0, UL9, US4), um gene estrutural (GAPDH) e um gene constitutivo (18S) da célula bovina tiveram suas expressões avaliadas por PCR quantitativa (qPCR). Enquanto os genes virais tiveram sua expressão aumentada ao longo do tempo de infecção, o gene hospedeiro teve sua expressão diminuída, demonstrando a ação do vírus na expressão gênica de células bovinas in vitro. O gene constitutivo 18S teve sua expressão mantida durante todos os momentos do experimento. Nossos resultados claramente demonstraram que o GAPDH não deve ser usado como gene de referência em estudos com infecção por BoHV-5 pois é influenciado pela infecção viral. Entretanto, o 18S rRNA foi constitutivamente expresso e pode ser recomendado para normalização em células bovinas infectadas pelo vírus. A expressão de mRNA viral não foi alterada pela quantidade de vírus usada. Todos os genes virais demonstraram o mesmo padrão de expressão ao longo do tempo de infecção. Nossos resultados trazem importantes diferenças comparando aos estudos clássicos que avaliaram a expressão de genes alfa, beta e gama. Mais estudos são necessários para aumentar o conhecimento da biologia molecular do BoHV-5. Estudo utilizando sequenciamento de última geração (i.e., RNA-seq), usando modelos in vitro e in vivo, aparentam ser o próximo passo lógico para acessar as alterações do transcriptoma do hospedeiro e viral ao longo do curso da infecção.(AU)

Correlation of feline immunodeficiency virus status and the severity of feline chronic gingivostomatitis / Correlação do status sorológico para FIV e a gravidade da gengivoestomatite crônica felina

Feline chronic gingivostomatitis (FCGS) is a challenge for the veterinary practitioner since its etiology and treatments are still undefined. The present paper investigated the role of the feline immunodeficiency virus (FIV) in the severity of the FCGS. Oral mucosal biopsies obtained from 19 cats with FCGS were divided into two groups according to their FIV serology status. Later, the clinical lesion score was correlated with the histopathological grade of FCGS lesions and the degree of immunostaining in both groups. Most of the animals had significant histological changes; however, no correlation with FIV immunostaining intensity was observed. It was concluded that the presence of FIV infection or the animal's seropositivity status does not seem to interfere with the severity of clinical signs nor the degree of histopathological changes when compared to the seronegative group.(AU)

A gengivoestomatite crônica felina (FCGS) é um desafio para o veterinário, uma vez que a sua etiologia e tratamentos permanecem indefinidos. O presente trabalho investigou o papel do vírus da imunodeficiência felina (FIV) na gravidade do FCGS. Biópsias da mucosa oral de 19 gatos com FCGS foram divididas em dois grupos de acordo com o status sorológico de FIV. Mais tarde, o escore de lesão clínica foi correlacionado com o grau histopatológico das lesões FCGS e o grau de imunocoloração em ambos os grupos. A maioria dos animais apresentou alterações histológicas significativas, porém não foi observada correlação com a intensidade de imunocoloração para FIV. Concluiu-se que a presença de infecção por FIV ou o estado soropositivo dos animais não parece interferir com a gravidade dos sinais clínicos nem com o grau de alterações histopatológicas quando comparado ao grupo soronegativo.(AU)

Mycobacterium genavense infection in two species of captive snakes

Background: Mycobacterium is an important zoonotic agent with companion, livestock and wildlife animals reportedly playing a role as reservoirs. Although its association with reptiles has been described, the disease cycle remains to be fully established, particularly in snakes. Accordingly, this study aimed to report the occurrence of mycobacteriosis with clinical pneumonia in one exotic python snake (Python molurus) and one native green snake (Philodryas olfersii) from the Sorocaba Zoo, São Paulo state, Brazil. Methods: Diagnosis was based on necropsy, histopathological examination, Ziehl-Neelsen stain and immunohistochemistry. Results: Using a nested PCR followed by DNA sequencing and bioinformatics analysis, the causative Mycobacterium species was identified as Mycobacterium genavense. Conclusion: Mycobacterium genavense is an infectious zoonotic agent of animal and public health concerns.(AU)

Mycobacterium genavense infection in two species of captive snakes

Mycobacterium is an important zoonotic agent with companion, livestock and wildlife animals reportedly playing a role as reservoirs. Although its association with reptiles has been described, the disease cycle remains to be fully established, particularly in snakes. Accordingly, this study aimed to report the occurrence of mycobacteriosis with clinical pneumonia in one exotic python snake (Python molurus) and one native green snake (Philodryas olfersii) from the Sorocaba Zoo, São Paulo state, Brazil. Methods: Diagnosis was based on necropsy, histopathological examination, Ziehl-Neelsen stain and immunohistochemistry. Results: Using a nested PCR followed by DNA sequencing and bioinformatics analysis, the causative Mycobacterium species was identified as Mycobacterium genavense. Conclusion: Mycobacterium genavense is an infectious zoonotic agent of animal and public health concerns.