Inflammation and changes in cytokine levels in neurological feline infectious peritonitis.

Feline infectious peritonitis (FIP) is a progressive, fatal, predominantly Arthus-type immune-mediated disease that is triggered when cats are infected with a mutant enteric coronavirus. The disease presents variably with multiple organ failure, seizures, generalized effusion, or shock. Neurological FIP is clinically and pathologically more homogeneous than systemic 'wet' or 'dry' FIP; thus, comparison of cytokine profiles from cats with neurological FIP, wet FIP, and non-FIP neurological disease may provide insight into some baseline characteristics relating to the immunopathogenesis of neurological FIP. This study characterizes inflammation and changes in cytokines in the brain tissue of FIP-affected cats. Cellular infiltrates in cats with FIP included lymphocytes, plasma cells, neutrophils, macrophages, and eosinophils. IL-1 beta, IL-6, IL-12, IL-18, TNF-alpha, macrophage inhibitory protein (MIP)-1 alpha, and RANTES showed no upregulation in the brains of control cats, moderate upregulation in neurological FIP cats, and very high upregulation in generalized FIP cats. Transcription of IFN-gamma appeared upregulated in cats with systemic FIP and slightly downregulated in neurological FIP. In most cytokines tested, variance was extremely high in generalized FIP and much less in neurological FIP. Principal components analysis was performed in order to find the least number of 'components' that would summarize the cytokine profiles in cats with neurological FIP. A large component of the variance (91.7%) was accounted for by levels of IL-6, MIP-1 alpha, and RANTES. These findings provide new insight into the immunopathogenesis of FIP and suggest targets for immune therapy of this disease

Desenvolvimento de um roteiro conceitual para a gestão da biodiversidade e dos serviços ecossistêmicos no Caribe mexicano

Coral reefs and mangroves support rich biodiversity and provide ecosystem services that range from food, recreational benefits and coastal protection services, among others. They are one of the most threatened ecosystems by urbanization processes. In this context, we developed a conceptual framework for the management of biodiversity and ecosystem services for these coastal environments. We based our workflow on two sections: “Information base” and “Governance” and use the Puerto Morelos Coastal region as a case study for coastal protection. Puerto Morelos is between two of the most touristic destinations of Mexico (Playa del Carmen and Cancun) that has experienced an increase of population in the past four decades resulting in an intensification of multiple threats to its ecosystems. We characterized the two ecosystems with a “Management Units” strategy. An expert-based ecosystem services matrix was also described in order to connect mangroves and coral reef ecosystems with the multiple beneficiaries. Then an ecosystem model (conceptual model and Global Biodiversity model) was developed. The conceptual model was useful in understanding the interplay processes between systems regarding the ecosystem service of “Coastal Protection”. The Global Biodiversity model evidenced the human-induced shifts in the biodiversity for mangrove and coral reefs ecosystems. Also, a projection for 2035 of “best” and “worst” scenarios was applied using GLOBIO3. A DPSIR conceptual framework was used to analyze environmental problems regarding ecosystem services maintenance. Finally, we evaluated a set of policies associated with these ecosystems that favor coastal protection integrity. This framework facilitates the identification of the most relevant processes and controls about the provision of coastal protection service. It can also be useful to better target management actions and as a tool to identify future management needs to tackle the challenges preventing more effective conservation of coastal environments.Recifes de coral e manguezais possuem rica biodiversidade e fornecem serviços ecossistêmicos, tais como, alimento, recreação, proteção costeira, entre outros. Esses ecossistemas encontram-se entre os mais ameaçados pelos processos de urbanização. Nesse contexto, desenvolvemos um roteiro conceitual para a gestão da biodiversidade e dos serviços ecossistêmicos desses ambientes costeiros. Organizamos nossa sequência de passos de trabalho em duas seções: “Base de informações” e “Governança” e usamos a região costeira da cidade de Puerto Morelos (México) como um estudo de caso para analisar o serviço de proteção de costa. Puerto Morelos encontra-se entre dois dos destinos mais turísticos do México (Playa del Carmen e Cancún), e portanto sua população vem aumentando nas últimas quatro décadas, resultando na intensificação de múltiplas ameaças para os ecossistemas. Primeiramente, caracterizamos os dois ecossistemas identificando-os como “Unidades de Gestão”, detalhando seus principais componentes e processos. Através de uma “Matriz de serviços ecossistêmicos”, construída com base na opinião de especialistas, foram sistematizados os principais serviços ecossistêmicos prestados pelos manguezais e recifes de corais aos múltiplos beneficiários. Em seguida, foi desenvolvida uma modelagem do sistema (e ecossistemas) através de sua representação na forma de um modelo conceitual e um modelo numérico de Biodiversidade Global. O modelo conceitual facilitou a compreensão dos processos de interação entre sistemas em relação ao serviço “Proteção Costeira”. O modelo numérico evidenciou as mudanças induzidas pelo homem na biodiversidade dos ecossistemas de manguezal e recifes de coral. Além disso, uma projeção dos cenários “melhor” e “pior” foi desenvolvida para 2035 usando GLOBIO3. A Estrutura conceitual DPSIR foi aplicada para analisar problemas ambientais relacionados à manutenção dos serviços ecossistêmicos. Finalmente, avaliamos um conjunto de políticas públicas associadas a esses ecossistemas e que favorecem a integridade da proteção costeira. Portanto, o roteiro facilitou a identificação dos principais processos e controles para a provisão de um serviço ecossistêmico. Além disso, pode ser útil para direcionar melhor as ações de gerenciamento, bem como, uma ferramenta para identificar necessidades futuras de planejamento e gestão para enfrentar desafios que permitam uma conservação mais eficaz dos ambientes costeiros.Fil: Sánchez Quinto, Andrés. Universidad Nacional Autónoma de México; MéxicoFil: Costa, Julliet Correa da. Universidade Federal de Santa Catarina; BrasilFil: Zamboni, Nadia Selene. Universidade Federal do Rio Grande do Norte; BrasilFil: Sanches, Fábio H. C.. Universidade Federal de Sao Paulo; BrasilFil: Principe, Silas C.. Universidade de Sao Paulo; BrasilFil: Viotto, Evangelina del Valle. Provincia de Entre Ríos. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción. Universidad Autónoma de Entre Ríos. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción; ArgentinaFil: Casagranda, Maria Elvira. Universidad Nacional de Tucumán. Instituto de Ecología Regional. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Ecología Regional; ArgentinaFil: Lima, Francisco A. da Veiga. Universidade Federal de Santa Catarina; BrasilFil: Possamai, Bianca. Universidade Federal Do Rio Grande.; BrasilFil: Faroni Perez, Larisse. Universidade Federal de Juiz de Fora; Brasi

Evaluation of real-time RT-PCR for the quantification of FCoV shedding in the faeces of domestic cats

Faecal samples were taken from cats living in multi-cat households with endemic feline coronavirus (FCoV) infection. Total RNA was extracted from faecal suspensions and FCoV RNA was quantified using a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay. The real-time RT-PCR threshold cycle (CT) values were consistently high suggesting that the samples contained very little viral RNA. However, experiments in which RNA extracted from FCoV-infected cell culture supernatants was combined with RNA extracted from faecal suspensions revealed the presence of faecal factors that significantly inhibited the reverse transcription reaction. Consequently, three methods of RNA extraction were investigated and RNA dilution was undertaken to investigate whether the effects of the faecal inhibitors could be reduced. Our results show that using the QIAgen RNA mini kit for RNA extraction and dilution of the RNA samples helps to reduce the inhibitory effects. However, because the extent of the inhibitory effects varied between faecal samples, accurate quantification proved difficult. We, therefore, conclude that although real-time RT-PCR provides an excellent method for detecting the presence of viral shedding, quantification of FCoV RNA in faecal material has to take into account the possible effects of RT-PCR inhibitors. It is, therefore, essential that all new assays, and the methods of sample preparation, are carefully evaluated before being used in a clinical setting

High viral loads despite absence of clinical and pathological findings in cats experimentally infected with feline coronavirus (FCoV) type I and in naturally FCoV-infected cats

Specified pathogen-free cats were naturally infected with FCoV or experimentally infected with FCoV type I. Seroconversion was determined and the course of infection was monitored by measuring the FCoV loads in faeces, whole blood, plasma and/or monocytes. Tissue samples collected at necropsy were examined for viral load and histopathological changes. Experimentally infected animals started shedding virus as soon as 2 days after infection. They generally displayed the highest viral loads in colon, ileum and mesenteric lymph nodes. Seroconversion occurred 3-4 weeks post infection. Naturally infected cats were positive for FCoV antibodies and monocyte-associated FCoV viraemia prior to death. At necropsy, most animals tested positive for viral shedding and FCoV RNA was found in spleen, mesenteric lymph nodes and bone marrow. Both experimentally and naturally infected cats remained clinically healthy. Pathological findings were restricted to generalized lymphatic hyperplasia. These findings demonstrate the presence of systemic FCoV infection with high viral loads in the absence of clinical and pathological signs